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Advances in Biotechnology

I. INTRODUCTION

The recent advances in science and technology have driven the biological sciences into a new era. All of this can probably be traced back to when Anton van Leeuwenhoek, a Dutch dry-goods dealer, ground the first microscope lens. Through his newly invented glass, he discovered a previously unseen cellular world. The second half of the 21st century was a truly exciting time for molecular biologists. Many inventions, including new methods for analyzing proteins, DNA and RNA, fuelled an explosion of information and enabled scientists to understand cells and multicellular organisms. The main objective of modern science, however, is to know the nature of genetic material and to find the answers to questions like: which genes determine specific characters? How do they get switched on and off spatially and temporally? How do we correct genetic defects? How can we best manipulate genomes?

A. What is Biotechnology?

The field of biotechnology, which emerged as a new discipline was a result of the fusion of biology and technology. Biology is the science of all living organisms or their components, whereas technology deals with the physical-chemical properties and techniques applied to the production of biological products/services. The emergence of biotechnology has been possible mainly due to the revolutionary discoveries made in these two areas. Biotechnology has been defined in many ways by many organizations. Biotechnology may be broadly defined as “the controlled use of selected/manipulated biological systems or processes for the production of abundant/novel products or service”. Therefore, the

area covered under biotechnology is very vast and the techniques involved are widely divergent. Biotechnology can be applied in areas as diverse as agriculture, animal husbandry, medicine, environment, industry, and biological conservation.

Biotechnology is multidisciplinary by its very nature and encompasses several disciplines of basic sciences (genetics, biochemistry, molecular

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Advances in Biotechnology

biology, chemistry, microbiology, immunology, cell and tissue culture, and physiology), and engineering (processing technology, biochemical engineering, electronics and physical sciences) and also other disciplines like sociology, economics, politics, law and ethics.

B. When did Biotechnology Begin?

Although the term biotechnology is a recent development, its origin can be traced back to prehistoric times. Humans have been altering the genetic composition of plants for millennia – retaining seeds from the best crops and planting them in the following years, breeding and cross-breeding varieties to make them taste sweeter, grow bigger, last longer, etc. in this way, early agriculturists transformed the wild tomato (Lycopersicon), from a fruit the size of a peanut to today’s giant, juicy and fleshy tomato. From a weedy plant called teosinte with an “ear” barely an inch long has emerged our foot-long ears of sweet, nutrient-rich, yellow corn. Man has also selected hundreds and thousands of new crop varieties by selection and hybridization. In ancient scripts, it has been documented that humans employed microorganisms as early as 5000 BC for making wine, vinegar, yogurt, leavened bread, etc. The discovery that fermentation converted fruit juice into wine, milk into cheese and yogurt, and solutions of malt and hops into beer seems to have set in motion the study of biotechnology. The early animal breeders soon realized that different physical traits could be either magnified or lost by mating the appropriate pairs of animals, thereby engaging in the traditional manipulations of biotechnology.

However, the use of microorganisms for the production of chemicals on a commercial scale begun during the First World War, and has recently been more fully exploited due to the advancement of modern biotechnology.

C. Modern Biotechnology

Modern biotechnology is innovative and quite different from the conventional practices. Traditional breeders made crosses only between related organisms whose genetic composition was compatible (genetically closer). Doing it this way involved the transfer of tens of thousands of genes (many genes were not required) after years of long selection procedure. By contrast, today’s genetic engineers can transfer just a few desirable genes at a time, between species that are distantly related or not related at all. In

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Advances in Biotechnology

other words, scientists can extract a desirable gene from virtually any living organism and insert it into virtually any other organism. They can put human gene into a plant or microorganism or in any other combination. For example, they can put a rat gene into lettuce to make a plant that produces vitamin C or insert a microbial toxin gene into cotton plants to make it insect-resistant. All this genetic manipulation became possible by the discovery of techniques of gene splicing and recombinant DNA technology. The engineered organisms which scientists produce by transforming genes between species are called “transgenic” organisms. Transgenic animals and several dozen transgenic food crops are currently in the market. Most of these crops are engineered to help farmers deal with age-old agriculture problems: good seeds, insects, diseases, nutrient composition, stress tolerance, etc.

The beginning of modern biotechnology can be traced back to 1865, when Gregor Mendel published the results from his experiments conducted on the garden pea on the inheritance of seven different physical traits. This and many other studies eventually led to the concept of the gene as the basic unit of heredity. Over the next century, many other researchers with sophisticated techniques and instruments contributed to the growth of modern biotechnology.

Before 6000 BC Yeasts used to make wine and beerAbout 4000 BC Yeasts were used for making leavened bread1866 Mendel published his research findings, experiments

conducted on the garden pea, which led to the concept of the gene as the basic unit of heredity.

1869 Friedrich Miescher isolated nuclein, later shown to be DNA, from the nuclei of white blood cells.

1885 E.Coli bacterial cells are identified and grown under controlled conditions.

1910 Thomas Hunt Morgan showed the first evidence of the presence of genes in chromosomes. He used microorganisms to treat sewage.

1912-14 Large scale production of acetone, butanol and glycerol using bacteria.

1917 Karl Ereky coined the term “biotechnology”1944 Avery, Macleod and McCarty demonstrated that DNA, not

protein; carries hereditary information. Penicillin was produced on large scale for the first time.

1957 The Central Dogma, which states that hereditary information flows from DNA to RNA to protein, was put

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forth by Francis Crick and George Gamov1981 The use of monoclonal antibodies for diagnosis was

approved in the USA.1990 Approval was granted in the USA for a trial of human

somatic cell gene therapy.To date The field of biotechnological research enormously

expanded and is still expanding exponentially.Table 1.1 Chronology of some major developments in biotechnology

D. Gene Technology is a Basic Tool for Biotechnology

Scientists continue to find new ways to insert desirable genes for specific traits into the DNA of different biological systems (plants, animals and microorganisms). A field of promise and a subject of debate, genetic engineering is changing the food we eat and the world we live in. Many scientist envision a cornucopia(similar to the “kalparuksha” or “Kamadenu”): the mass production of rare plants; highly variegated and long shelf-life flowers; tomatoes and broccoli produced with pharmaceutical compounds and industrial chemicals; vaccine-producing bananas; vitamin-enriched rice, sweet potatoes, and cassava to help the malnourished poor and vegetable oils so loaded with therapeutic ingredients that doctors “prescribe” them for patients at risk of cancer, heart disease, diabetes, etc; cheaper and safer fuel; clean environment, etc. The possibilities are endless.

Overall, gene manipulation has provided novel solutions to experimental problems in biology; these solutions, in turn, have led to novel products. Most biotechnology companies and research institutes make use of gene technology or genetic engineering, which mainly involves recombinant DNA and gene cloning. This technology allows the splicing of a DNA molecule at desired places to isolate specific DNA segment and then inserting it into another DNA molecule at a desired position. The resultant product is called “recombinant DNA” and the technique often called “genetic engineering”. Using molecular techniques, we can isolate and clone a single copy of a gene or a DNA segment into an indefinite number of copies with similar properties. This became possible due to the identification and modification of different kinds of vectors(a self-replicating DNA molecule is called “vector” e.g., plasmid, phage or virus) with suitable properties, and which can be mobilized to a suitable host, where they reproduce along with the host. The vector carrying the inserted DNA will also replicate faithfully through the vector DNA. This technique is called “gene cloning”. A gene is a part of a chromosome and is responsible for a specific character or trait of

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Advances in Biotechnology

an organism/cell. Genes produce their phenotypic effects by specifying the amino acid sequences of specific proteins. There are also various biological tools that are used to carry out manipulation of genetic material and cells.

The gene cloning technique has had a tremendous impact on all areas of molecular biology and, consequently, on biotechnology. Recombinant DNA technology broadly involves: (1) the isolation of a specific DNA, (2) the selection of vectors, (3) the preparation of a chimeric DNA, (5) cloning of the chimeric DNA, and (6) the screening of recombinants.

II. WHAT MAKES BIOTECHNOLOGY A POWERFUL TECHNOLOGY?

Biotechnology is a unique and powerful technology, since it helps humans in many ways, for example, (i) mass production, (ii) generation of novelty, and (iii) better service.

A. Overproduction of Cellular Components

For commercial use or in academic studies, the determination of the structure, function or utility of a protein demands that adequate amounts of purified material are available. This is not always an easy task, particularly when the protein is normally present in very low levels in the cell mass. Genetic engineering provides a means of generating sufficient material. For example, 5 mg of somatostatin was first isolated from half a million sheep brains and a small amount of epidermal growth factor from 40,000 gallons of human urine. After the advent of gene cloning, the same amount of material was obtained from a few litters of bacterial culture. This principle has been applied to a wide range of cellular proteins and was the basis for many of the biotechnology start-up companies in the world. Over-production need not be restricted to proteins. It is possible to raise the levels of most intracellular components, provided that they are not toxic to the producing organism. This can be done by cloning all the genes for a particular biosynthetic pathway and over-expressing them. Alternatively, it is possible to shut down particular metabolic pathways and thus re-direct particular inter mediates towards the desired end product.

B. Generation of Novelty

By and large, gene manipulation has provided novel solutions to experimental problems in biology. These solutions have led to creation of novel products. Gene manipulation has been used to permanently modify

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the germ cells of animals (“transgenesis”), e.g., the production of ‘supermice’ which are extra-large as a result of the over-production of the human growth hormone. Transgenic animals that over-express foreign proteins and secrete them in milk have been developed. Novel therapies for various human diseases have been also created. Transgenic plants carrying genes resistant to various stresses have also been produced. However, the development of these products has also raised some novel problems. Some of these benefits have been discussed in the following sections.

C. Better ServiceBiotechnology can also be of better service to human beings, other

organisms and to the ecosystem by providing solutions to many natural and man-made problems. For example, biotechnology can provide solutions to environmental pollution; it can help to monitor and conserve biodiversity, and reduce the rapid loss of natural resources by providing alternative solutions. It can also aid in the overall improvement of biological safety and in maintaining the ecological balance.

III. APPLICATIONS OF BIOTECHNOLOGY

Biotechnology has rapidly emerged as an area of activity that will have a potential impact on virtually all domains of human welfare; food processing, human health, agriculture animal improvement, and environmental protection. As a result, it plays a very important role in employment, production, trade, economics and economy, human health, conservation of biodiversity and even in the socio-economical and political status of a nation. This is clearly reflected in the emergence of numerous biotechnology companies throughout the world.

The importance of biotechnology to human welfare is becoming increasingly more important. The products of DNA research, ranging from proteins to engineering organisms, have a wide range of applications. The following are some of the contributions of biotechnology to overall development of human welfare.

A. Medical Biotechnology

1. Diagnosis of biological disorders

The production of biological reagents for the diagnosis of biological disorders and infectious diseases is a major industry. On a global scale, the

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revenue from sales is estimated at many billions of dollars. There are three types of diagnostic reagents: biochemical reagents for assaying specific enzymes, antibodies for detecting specific proteins, and a recent development, nucleic probes.

Molecular analysis of genetic disorders: There are several hundred genetic diseases in man, which are the result of mutations. For many of these genetic diseases there is no definitive treatment, although in some cases human gene therapy may become possible. DNA research helps in understanding many diseases or genetic disorders at the molecular level, such as sickle cell anemia, thalassemias, familial hypercholesterolemia, etc.

Laboratory diagnostic applications: By using recombinant DNA techniques, many diseases can be diagnosed, e.g., AIDS, hepatitis B, etc. In the case of most diseases, diagnosis is the detection of a specific microorganism. For example, a patient with a disorder of the gastrointestinal tract could be infected with more than 15 types of microorganisms. By using the right probe, a particular infectious microbe can be accurately detected from stool samples without any cultivation and cytological work.

Prenatal diagnosis of diseases: The DNA collected from the amniotic fluid can be used to predict the risk of developing genetic diseases (e.g., sickle cell anemia and many other genetic defects) and this can be done by using DNA probes.

2. Hybridoma technology

This produces large quantities of monoclonal antibodies, which are used for the diagnosis of various diseases, e.g., venereal diseases, hepatitis B, viral diseases, cancer, etc.

3. Genetically-engineered microbes

Vaccines: conventional viral vaccines consist of inactivated, virulent strains or live, attenuated strains, but they are not without their problems (e.g., there is a danger of vaccine-related disease). Many problems can be overcome by recombinant vaccines, which are cleaner and safer (e.g., human hepatitis B virus, E. coli vaccines for pigs, rabbits, etc.)

GEMs can be used to produce DNA probes that can be used for diagnosing diseases such as ka-azar, sleeping sickness, malaria, etc. They

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can also be a source of valuable drugs like human insulin, human interferon, human and bovine growth hormones, etc.

4. Advanced therapeutic techniques

Production of proteins in abundance: Using recombinant DNA techniques, several proteins have been produced in abundance for therapeutic purposes. These include insulin, growth hormone, erythropoietin, interferons, blood-clotting factors, vaccines and superoxide dismutase.

Artificial insemination and sexing: genetic procedures can be used to trea infertility as well as produce babies of a specified sex (by artificial insemination with X or Y carrying sperms, prepared by sperm separation techniques).

5. Application to forensic medicine

Finger-printing and forensics: in paternity disputes, the parents can be identified by using DNA or auto-antibody finger-printing. Using the same technique, criminals can be identified very accurately by blood or semen stains, hair roots, etc., collected from a crime scene and compared with the suspect’s DNA.

B. Animal Biotechnology

1. In vitro fertilization and embryo transfer

In vitro fertilization and embryo transfer have been successfully applied both in humans and in animals. In the case of human beings with the help of biotechnology, some couples suffering from infertility can bear children.

The rapid multiplication of superior genotypes in animals can be achieved by hormone-induced superovulation, in vitro fertilization and/or embryo splitting followed by embryo transfer into a foster mother.

2. Transgenic animals

Transgenic animals can be created for increased milk, rapid growth rate, resistance to diseases and the production of valuable proteins in the milk/serum/urine. Many transgenic animals carrying novel characteristics have been developed in mice, pigs, chicken, rabbits, cattle, sheep and fish.

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Transgenic fish: Atlantic salmon grow more slowly during the winter, but engineered salmon carrying modified growth-hormone genes from other fish, reach market size in about half the normal time.

Pharmaceutical-producing animals: Scientists are using biotechnology to insert genes into cows and sheep, so that the animals produce beneficial pharmaceuticals in their milk.

C. Plant Biotechnology

1. Diagnostics

The accurate identification of viruses is critical in the prediction of plant diseases in annual crops, for the prevention of infection in planting stock, in monitoring disease-control methods and in diagnosing diseases in plants held in quarantine. Biotechnology is a useful tool in identifying viruses.

2. Plant cell culture

Clonal propagation: A very high proportion of plant material can be multiplied by clonal propagation through meristem culture. For example, many fruit and forest trees are very slow-growing and their growth can be increased manifold by means of biotechnological protocols.

Embryo culture: Embryo culture is used to rescue ubviable hybrids and haploid plants from inter-specific hybrids. This is one of the most important techniques in plant breeding programs.

Virus-free plants: Meristem culture is generally combined with thermotherapy/cryotherapy. This procedure is usually followed to recover virus- and pathogen-free stocks of clonal crops. It is very useful in clonal crops and germplasm exchange.

Anther culture: The rapid isolation of homozygous lines can be achieved by chromosome doubling of haploids produced through anther culture. This technique has been successfully used in the development of variety, e.g., in rice and wheat.

Somaclonal variation: This refers to the isolation of stable somaclonal variants with improved yield and other desirable traits such as resistance to diseases, cold, herbicides, metal toxicity, salt and other abiotic stresses.

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3. Transgenic plants

Transgenic plants have been established in many crop varieties by using transfer techniques for various characteristics, which include insect resistance, protection against viruses, herbicide resistance, storage protein improvement, secondary metabolites, etc.

Herbicide resistance: Farmers routinely spray herbicides to kill weeds. However, biotech crops can carry special “tolerance” genes that help them withstand the spraying of chemicals that kill nearly every other kind of plant.

Insect resistance: Some biotech varieties make their own insecticides. For example, Bt transgenics, which is a gene borrowed from a common soil bacterium, Bacillus thuringiensis, (Bt for short). Bt genes code for toxins that are considered harmless to humans but lethal to certain insects. When these insects feed on Bt plants, the toxin attacks their digestive tracts, and they die within a few days.

Disease resistance: Many crops have been genetically engineered to resist diseases. For example, squash and papaya have been genetically engineered to resist viral diseases. Potatoes have been transformed with the genes of bees and moths to protect the crops from the potato blight fungus, and grapevines have been altered with silkworm genes to make the vines resistant to Pierce’s disease, which is spread by insects.

Generation of novel food content of plants: It is widely recognized that gene manipulation techniques have revitalized the biotechnology industry. Altered starch content, oil and amino acid composition in storage proteins of seeds are a few of the many transgenic food crops developed.

Pigmentation in transgenic plants: Plants are widely used for ornamental purposes; therefore, considerable attempts have been made to develop varieties exhibiting new colors or pigmentation patterns. The pigmentation in flowers is mainly due to three classes of compounds: the flavonoids, the carotenoids and the betalains. Several flavonoids genes have been cloned and many-hued color patterns have been created.

4. Molecular mapping

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Molecular markers have been extensively used both for linkage mapping and for the mapping of quantitative trait loci. These techniques are very powerful tools for the indirect selection of quantitative traits and for several other important applications.

D. Industrial Biotechnology

Industrial Biotechnology is also known as white biotechnology. It is biotechnology applied to industrial processes. White biotechnology tends to consume less in resources than traditional processes used to produce industrial goods. The investment and economic output of all of these types of applied biotechnologies is termed as bioeconomy.

Microorganisms have been identified and exploited for more than a century. The Babylonians and Sumerians used yeast to prepare alcohol. There is a great history beyond fermentation processes, which explains the applications of microbial processes that resulted in the production of food and beverages. In the mid-nineteenth century, Louis Pasteur understood the role of microorganisms in fermented food, wine, alcohols, beverages, cheese, milk, yoghurt and other dairy products, fuels, and fine chemical industries. He identified many microbial processes and discovered the first principal role of fermentation, which was that microbes required substrate to produce primary and secondary metabolites, and end products.

In the new millennium, extensive application of bioprocesses has created an environment for many engineers to expand the field of biotechnology. One of the useful applications of biotechnology is the use of microorganisms to produce alcohols and acetone, which are used in the industrial processes. The knowledge related to industrial microbiology has been revolutionised by the ability of genetically engineered cells to make many new products.Genetic engineering and gene mounting have been developed in the enhancement of industrial fermentation. Consequently, biotechnology is a new approach to making commercial products by using living organisms. Furthermore, knowledge of bioprocesses has been developed to deliver fine-quality products. Application of biological sciences in industrial processes is known as bioprocessing. Nowadays most biological and pharmaceutical products are produced in well-defined industrial bioprocesses. For instance, bacteria are able to produce most amino acids that can be used in food and medicine. There are hundreds of microbial and fungal products purely available in the biotechnology market.

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An industrial biotechnology process that uses microorganisms for producing a commercial product typically has three operational stages.

1. Upstream Processing is the preparation of the raw materials so that it can be used as a food source for the target microorganism.

2. Fermentation and Transformation is the growth of the desirable microorganism in a fermenter (usually a bioreactor >100 liters), which produces the desired product by fermentation/biotransformation process.

3. Downstream Processing involves the purification of the desired product either from the cell medium or the cell-mass.

Production of Organic Compounds

Lactic AcidSeveral carbohydrates such as corn and potato starch, molasses and

whey can be used to produce lactic acid. Starch must first be hydrolysed to glucose by enzymatic hydrolysis; then fermentation is performed in the second stage. The choice of carbohydrate material depends upon its availability, and pretreatment is required before fermentation. We shall describe the bioprocess for the production of lactic acid from whey. Large quantities of whey constitute a waste product in the manufacture of dairy

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products such as cheese. From the standpoint of environmental pollution it is considered a major problem, and disposal of untreated wastes may create environmental disasters.

It is desirable to use whey to make some more useful product. Whey can be converted from being a waste product to something more desirable that can be used for the growth of certain bacteria, because it contains lactose, nitrogenous substances, vitamins and salts. Organisms can utilise lactose and grow on cheese wastes; the most suitable of them are Lactobacillus species such as Lactobacillus bulgaricus, which is the most suitable species for whey. This organism grows rapidly, is homofermentative and thus capable of converting lactose to the single end-product of lactic acid. Stock cultures of the organism are maintained in skimmed milk medium. The 3–5% of inoculum is prepared and transferred to the main bioreactor, and the culture is stored in pasteurised, skimmed milk at an incubation temperature of 43 °C.

During fermentation, pH is controlled by the addition of slurry of lime to neutralize the product to prevent any product inhibition. The accumulation of lactic acid would retard the fermentation process because of the formation of calcium lactate. After 2 days of complete incubation, the material is boiled to coagulate the protein, and then filtered. The solid filter cake is a useful, enriched protein product, which may be used as an animal feed supplement. The filtrate containing calcium lactate is then concentrated by removing water under vacuum, followed by purification of the final product.

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Acetic AcidThe sugars in fruits such as grapes are fermented by yeasts to

produce wines. In winemaking, lactic acid bacteria convert malic acid into lactic acid in malolactic fermentation in fruits with high acidity. Acetobacter and Gluconobacter oxidise ethanol in wine to acetic acid (vinegar). The word ‘wine’ is derived from the French term ‘vinaigre’ meaning ‘sour wine’. It is prepared by allowing a wine to get sour under controlled conditions. The production of vinegar involves two steps of biochemical changes:

(1) Alcoholic fermentation in fermentation of a carbohydrate.(2) Oxidation of the alcohol to acetic acid.

There are several kinds of vinegar. The differences between them are primarily associated with the kind of material used in the alcoholic fermentation, e.g. fruit juices, sugar and hydrolysed starchy materials. Based on US Department of Agriculture (USDA) definitions, there are a few types of vinegar: vinegar, cider vinegar, apple vinegar. The products are made by the alcoholic and subsequent acetous fermentations of the apple juice. The acetic acid content is about 5%. Yeast fermentation is used for the production of alcohol. The alcohol is adjusted to 10–13%, then it is exposed to acetic acid bacteria (Acetobacter species), whereby oxygen is

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required for the oxidation of alcohol to acetic acid. The desired temperature for Acetobacter is 15–34 °C. The reaction is:

Amino acids (lysine and glutamic acid) and insulinMany microorganisms can synthesise amino acids from inorganic

nitrogen compounds. The rate and amount of some amino acids may exceed the cells’ need for protein synthesis, where the excess amino acids are excreted into the media. Some microorganisms are capable of producing certain amino acids such as lysine, glutamic acid and tryptophan.

o Stepwise Amino Acid ProductionOne of the commercial methods for production of lysine consists of a two-stage process using two species of bacteria. The carbon sources for production of amino acids are corn, potato starch, molasses, and whey. If starch is used, it must be hydrolysed to glucose to achieve higher yield. Escherichia coli is grown in a medium consisting of glycerol, cornsteep liquor and di-ammonium phosphate under aerobic conditions, with temperature and pH controlled.

• Step 1: Formation of diaminopimelic acid (DAP) by E. coli.• Step 2: Decarboxylation of DAP by Enterobacter aerogenes.

E. coli can easily grow on corn steep liquor with phosphate buffer for an incubation period of 3 days. Lysine is an essential amino acid for the nutrition of humans, which is used as a supplementary food with bread and other foodstuffs. This amino acid is a biological product which is also used as a food additive and cereal protein.

Many species of microorganisms, especially bacteria and fungi, are capable of producing large amounts of glutamic acid. Glutamic acid is produced by microbial metabolites of Micrococcus, Arthrobacter, and Brevibacterium by the Krebs cycle. Monosodium glutamate is known as a flavour-enhancing amino acid in food industries. The medium generally used consists of carbohydrate, peptone, inorganic salts and biotin. The concentration of biotin has a significant influence on the yield of glutamic

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acid. The _-ketoglutaric acid is an intermediate in the Krebs cycle and is the precursor of glutamic acid. The conversion of _-ketoglutaric acid to glutamic acid is accomplished in the presence of glutamic acid dehydrogenase, ammonia and nicotinamide adenine dinucleotide dehydrogenase (NADH2). The living cells assimilate nitrogen by incorporating it into ketoglutaric acid, then to glutamic acid and glutamine. Therefore glutamic acid is formed by the reaction between ammonia and _-ketoglutaric acid in one of the tricarboxylic acid (TCA) cycle or Krebs cycle intermediates.

Insulin is one of the important pharmaceutical products produced commercially by genetically engineered bactera. Before this development, commercial insulin was isolated from animal pancreatic tissue. Microbial insulin has been available since 1982. The human insulin gene is introduced into a bacterium like E. coli. Two of the major advantages of insulin production by microorganisms are that the resultant insulin is chemically identical to human insulin, and it can be produced in unlimited quantities.

PenicillinThe commercial production of penicillin and other antibiotics are the

most dramatic in industrial microbiology. The annual production of bulk penicillin is about 33 thousand metric tonnes with annual sales market of more than US$400 million.8 The worldwide bulk sales of the four most important groups of antibiotics, penicillins, cephalosporins, tetracyclines and erythromycin, are US$4.2 billion per annum. The mold isolated by Alexander Fleming in early 1940s was Penicillium notatum, who noted that this species killed his culture of Staphylococcus aureus. The production of penicillin is now done by a better penicillin-producing mould species, Penicillium chrysogenum. Development of submerged culture techniques enhanced the cultivation of the mould in large-scale operation by using a sterile air supply.

• Streptomycin produced by Actinomycetes• Molasses, corn steep liquor, waste product from sugar industry, and

wet milling corn are used for the production of penicillin• Penicillium chrysogenum can produce 1000 times more penicillin

than Fleming’s original culture• The major steps in the commercial production of penicillin are:(1) Preparation of inoculum.(2) Preparation and sterilisation of the medium.(3) Inoculation of the medium in the fermenter.

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(4) Forced aeration with sterile air during incubation.(5) Removal of mould mycelium after fermentation.(6) Extraction and purification of the penicillin.

BiotransformationThis is the transformation of toxic compounds to less toxic or non-

toxic compounds. By means of biotransformation, less useful and cheaper compounds can be converted into more useful and valuable ones using microorganisms or immobilized enzyme in a fermenter.

Biotransformation is the chemical modification (or modifications) made by an organism on a chemical compound. If this modification ends in mineral compounds like CO2, NH4

+ or H2O, the biotransformation is called mineralisation. Biotransformation means chemical alteration of chemicals such as (but not limited to) nutrients, amino acids, toxins, or drugs in the body. It is also needed to render nonpolar compounds polar so that they are not reabsorbed in renal tubules and are excreted.

Drug metabolism

The metabolism of a drug or toxin in a body is an example of a biotransformation. Typically the body deals with a foreign compound by making it more soluble, to increase the rate of its excretion through the urine. There are a number of different process that can occur; the pathways of drug metabolism can be divided into:

phase І phase II

Drugs can undergo one of four potential biotransformations: Active Drug to Inactive Metabolite, Active Drug to Active Metabolite, Inactive Drug to Active Metabolite, Active Drug to Toxic Metabolite (biotoxification).

Phase I reaction

Includes oxidative, reductive and hydrolytic reactions. In these type of reactions, a polar group is either introduced or unmasked,

so the drug molecule becomes more water-soluble and can be excreted. Reactions are non-synthetic in nature & generally produce a more water

soluble & less active metabolites.

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The majority of metabolites are generated by a common hydroxylating enzyme system known as Cytochrome P450.

Phase II reaction

These reactions involve covalent attachment of small polar endogenous molecule such as glucuronic acid, sulfate, or glycine to form water-soluble compounds.

This is also known as a conjugation reaction.

The final compounds have a larger molecular weight.

Microbial biotransformation

Biotransformation of various pollutants is a sustainable way to clean up contaminated environments.[1] These bioremediation and biotransformation methods harness the naturally occurring, microbial catabolic diversity to degrade, transform or accumulate a huge range of compounds including hydrocarbons (e.g. oil), polychlorinated biphenyls (PCBs), polyaromatic hydrocarbons (PAHs), pharmaceutical substances, radionuclides and metals. Major methodological breakthroughs in recent years have enabled detailed genomic, metagenomic, proteomic, bioinformatic and other high-throughput analyses of environmentally relevant microorganisms providing unprecedented insights into biotransformation and biodegradative pathways and the ability of organisms to adapt to changing environmental conditions.

Biological processes play a major role in the removal of contaminants and pollutants from the environment. Some microorganisms possess an astonishing catabolic versatility to degrade or transform such compounds. New methodological breakthroughs in sequencing, genomics, proteomics, bioinformatics and imaging are producing vast amounts of information. In the field of Environmental Microbiology, genome-based global studies open a new era providing unprecedented in silico views of metabolic and regulatory networks, as well as clues to the evolution of biochemical pathways relevant to biotransformation and to the molecular adaptation strategies to changing environmental conditions. Functional genomic and metagenomic approaches are increasing our understanding of the relative importance of different pathways and regulatory networks to carbon flux in particular environments and for particular compounds and they are

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accelerating the development of bioremediation technologies and biotransformation processes. Also there is other approach of biotransformation called enzymatic biotransformation.

Metabolic Engineering and Biocatalytic Applications

The study of the fate of persistent organic chemicals in the environment has revealed a large reservoir of enzymatic reactions with a large potential in preparative organic synthesis, which has already been exploited for a number of oxygenases on pilot and even on industrial scale. Novel catalysts can be obtained from metagenomic libraries and DNA sequence based approaches. Our increasing capabilities in adapting the catalysts to specific reactions and process requirements by rational and random mutagenesis broadens the scope for application in the fine chemical industry, but also in the field of biodegradation. In many cases, these catalysts need to be exploited in whole cell bioconversions or in fermentations, calling for system-wide approaches to understanding strain physiology and metabolism and rational approaches to the engineering of whole cells as they are increasingly put forward in the area of systems biotechnology and synthetic biology.

Biomass Production

Biomass, a renewable energy source, is biological material from living, or recently living organisms, such as wood, waste, (hydrogen) gas, and alcohol fuels. Biomass is commonly plant matter grown to generate electricity or produce heat. In this sense, living biomass can also be included, as plants can also generate electricity while still alive. The most conventional way in which biomass is used however, still relies on direct incineration. Forest residues for example (such as dead trees, branches and tree stumps), yard clippings, wood chips and garbage are often used for this. However, biomass also includes plant or animal matter used for production of fibers or chemicals. Biomass may also include biodegradable wastes that can be burnt as fuel. It excludes such organic materials as fossil fuels, which have been transformed by geological processes into substances such as coal or petroleum.

Industrial biomass can be grown from numerous types of plants, including miscanthus, switchgrass, hemp, corn, poplar, willow, sorghum,

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sugarcane, and a variety of tree species, ranging from eucalyptus to oil palm (palm oil). The particular plant used is usually not important to the end products, but it does affect the processing of the raw material.

Although fossil fuels have their origin in ancient biomass, they are not considered biomass by the generally accepted definition because they contain carbon that has been "out" of the carbon cycle for a very long time. Their combustion therefore disturbs the carbon dioxide content in the atmosphere.

Chemical composition

Biomass is carbon, hydrogen and oxygen based. Nitrogen and small quantities of other atoms, including alkali, alkaline earth and heavy metals can be found as well. Metals are often found in functional molecules such as the porphyrins which include chlorophyll which contains magnesium.

Plants in particular combine water and carbon dioxide to sugar building blocks. The required energy is produced from light via photosynthesis based on chlorophyll. On average, between 0.1 and 1 % of the available light is stored as chemical energy in plants. The sugar building blocks are the starting point for the major fractions found in all terrestrial plants, lignin, hemicellulose and cellulose.[4]

Biomass sources

Biomass energy is derived from five distinct energy sources: garbage, wood, waste, landfill gases, and alcohol fuels. Wood energy is derived both from direct use of harvested wood as a fuel and from wood waste streams. The largest source of energy from wood is pulping liquor or “black liquor,” a waste product from processes of the pulp, paper and paperboard industry. Waste energy is the second-largest source of biomass energy. The main contributors of waste energy are municipal solid waste (MSW), manufacturing waste, and landfill gas. Biomass alcohol fuel, or ethanol, is derived primarily from sugarcane and corn. It can be used directly as a fuel or as an additive to gasoline.

Biomass can be converted to other usable forms of energy like methane gas or transportation fuels like ethanol and biodiesel. Rotting garbage, and agricultural and human waste, release methane gas - also called "landfill gas" or "biogas." Crops like corn and sugar cane can be

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fermented to produce the transportation fuel, ethanol. Biodiesel, another transportation fuel, can be produced from left-over food products like vegetable oils and animal fats. Also, Biomass to liquids (BTLs) and cellulosic ethanol are still under research.

Biomass conversion process to useful energy

There are a number of technological options available to make use of a wide variety of biomass types as a renewable energy source. Conversion technologies may release the energy directly, in the form of heat or electricity, or may convert it to another form, such as liquid biofuel or combustible biogas. While for some classes of biomass resource there may be a number of usage options, for others there may be only one appropriate technology.

Thermal conversion

These are processes in which heat is the dominant mechanism to convert the biomass into another chemical form. The basic alternatives of combustion, torrefaction, pyrolysis, and gasification are separated principally by the extent to which the chemical reactions involved are allowed to proceed (mainly controlled by the availability of oxygen and conversion temperature).

There are a number of other less common, more experimental or proprietary thermal processes that may offer benefits such as hydrothermal upgrading (HTU) and hydroprocessing. Some have been developed for use on high moisture content biomass, including aqueous slurries, and allow them to be converted into more convenient forms. Some of the applications of thermal conversion are combined heat and power (CHP) and co-firing. In a typical biomass power plant, efficiencies range from 20-27%.[9]

Chemical conversion

A range of chemical processes may be used to convert biomass into other forms, such as to produce a fuel that is more conveniently used, transported or stored, or to exploit some property of the process itself.

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Biochemical conversion

As biomass is a natural material, many highly efficient biochemical processes have developed in nature to break down the molecules of which biomass is composed, and many of these biochemical conversion processes can be harnessed.

Biochemical conversion makes use of the enzymes of bacteria and other micro-organisms to break down biomass. In most cases micro-organisms are used to perform the conversion process: anaerobic digestion, fermentation and composting. Other chemical processes such as converting straight and waste vegetable oils into biodiesel is transesterification. Another way of breaking down biomass is by breaking down the carbohydrates and simple sugars to make alcohol. However, this process has not been perfected yet. Scientists are still researching the effects of converting biomass.

Biofuels

Biofuels are a wide range of fuels which are in some way derived from biomass. The term covers solid biomass, liquid fuels and various biogases.[1]

Biofuels are gaining increased public and scientific attention, driven by factors such as oil price spikes, the need for increased energy security, and concern over greenhouse gas emissions from fossil fuels.

Liquid fuels for transportation

Most transportation fuels are liquids, because vehicles usually require high energy density, as occurs in liquids and solids. High power density can be provided most inexpensively by an internal combustion engine; these engines require clean burning fuels, to keep the engine clean and minimize air pollution.

The fuels that are easiest to burn cleanly are typically liquids and gases. Thus liquids (and gases that can be stored in liquid form) meet the requirements of being both portable and clean burning. Also, liquids and gases can be pumped, which means handling is easily mechanized, and thus less laborious.

First generation biofuels

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'First-generation biofuels' are biofuels made from sugar, starch, and vegetable oil.

Bioalcohols

Biologically produced alcohols, most commonly ethanol, and less commonly propanol and butanol, are produced by the action of microorganisms and enzymes through the fermentation of sugars or starches (easiest), or cellulose (which is more difficult). Biobutanol (also called biogasoline) is often claimed to provide a direct replacement for gasoline, because it can be used directly in a gasoline engine (in a similar way to biodiesel in diesel engines).

Ethanol fuel is the most common biofuel worldwide, particularly in Brazil. Alcohol fuels are produced by fermentation of sugars derived from wheat, corn, sugar beets, sugar cane, molasses and any sugar or starch that alcoholic beverages can be made from (like potato and fruit waste, etc.). The ethanol production methods used are enzyme digestion (to release sugars from stored starches), fermentation of the sugars, distillation and drying. The distillation process requires significant energy input for heat (often unsustainable natural gas fossil fuel, but cellulosic biomass such as bagasse, the waste left after sugar cane is pressed to extract its juice, can also be used more sustainably).

Ethanol can be used in petrol engines as a replacement for gasoline; it can be mixed with gasoline to any percentage. Most existing car petrol engines can run on blends of up to 15% bioethanol with petroleum/gasoline. Ethanol has a smaller energy density than gasoline, which means it takes more fuel (volume and mass) to produce the same amount of work. An advantage of ethanol (CH3CH2OH) is that it has a higher octane rating than ethanol-free gasoline available at roadside gas stations which allows an increase of an engine's compression ratio for increased thermal efficiency. In high altitude (thin air) locations, some states mandate a mix of gasoline and ethanol as a winter oxidizer to reduce atmospheric pollution emissions.

Ethanol is also used to fuel bioethanol fireplaces. As they do not require a chimney and are "flueless", bio ethanol fires are extremely useful for new build homes and apartments without a flue. The downside to these fireplaces, is that the heat output is slightly less than electric and gas fires.

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In the current alcohol-from-corn production model in the United States, considering the total energy consumed by farm equipment, cultivation, planting, fertilizers, pesticides, herbicides, and fungicides made from petroleum, irrigation systems, harvesting, transport of feedstock to processing plants, fermentation, distillation, drying, transport to fuel terminals and retail pumps, and lower ethanol fuel energy content, the net energy content value added and delivered to consumers is very small. And, the net benefit (all things considered) does little to reduce un-sustainable imported oil and fossil fuels required to produce the ethanol.

Although ethanol-from-corn and other food stocks has implications both in terms of world food prices and limited, yet positive energy yield (in terms of energy delivered to customer/fossil fuels used), the technology has led to the development of cellulosic ethanol. According to a joint research agenda conducted through the U.S. Department of Energy, the fossil energy ratios (FER) for cellulosic ethanol, corn ethanol, and gasoline are 10.3, 1.36, and 0.81, respectively.

Many car manufacturers are now producing flexible-fuel vehicles (FFV's), which can safely run on any combination of bioethanol and petrol, up to 100% bioethanol. They dynamically sense exhaust oxygen content, and adjust the engine's computer systems, spark, and fuel injection accordingly. This adds initial cost and ongoing increased vehicle maintenance. As with all vehicles, efficiency falls and pollution emissions increase when FFV system maintenance is needed (regardless of the fuel mix being used), but is not performed. FFV internal combustion engines are becoming increasingly complex, as are multiple-propulsion-system FFV hybrid vehicles, which impacts cost, maintenance, reliability, and useful lifetime longevity.

Even dry ethanol has roughly one-third lower energy content per unit of volume compared to gasoline, so larger / heavier fuel tanks are required to travel the same distance, or more fuel stops are required. With large current unsustainable, non-scalable subsidies, ethanol fuel still costs much more per distance traveled than current high gasoline prices in the United States.

Methanol is currently produced from natural gas, a non-renewable fossil fuel. It can also be produced from biomass as biomethanol. The methanol economy is an interesting alternative to get to the hydrogen

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economy, compared to today's hydrogen production from natural gas. But this process is not the state-of-the-art clean solar thermal energy process where hydrogen production is directly produced from water.

Butanol is formed by ABE fermentation (acetone, butanol, ethanol) and experimental modifications of the process show potentially high net energy gains with butanol as the only liquid product. Butanol will produce more energy and allegedly can be burned "straight" in existing gasoline engines (without modification to the engine or car), and is less corrosive and less water soluble than ethanol, and could be distributed via existing infrastructures. DuPont and BP are working together to help develop Butanol. E. coli have also been successfully engineered to produce Butanol by hijacking their amino acid metabolism.

Fermentation is not the only route to forming biofuels or bioalcohols. One can obtain methanol, ethanol, butanol or mixed alcohol fuels through pyrolysis of biomass including agricultural waste or algal biomass. The most exciting of these pyrolysis alcoholic fuels is the pyrolysis biobutanol. The product can be made with limited water use and most places in the world.

Green diesel

Green diesel, also known as renewable diesel, is a form of diesel fuel which is derived from renewable feedstock rather than the fossil feedstock used in most diesel fuels. Green diesel feedstock can be sourced from a variety of oils including canola, algae, jatropha and salicornia in addition to tallow. Green diesel uses traditional fractional distillation to process the oils, not to be confused with biodiesel which is chemically quite different and processed using transesterification.

“Green Diesel” as commonly known in Ireland should not be confused with dyed green diesel sold at a lower tax rate for agriculture purposes, using the dye allows custom officers to determine if a person is using the cheaper diesel in higher taxed applications such as commercial haulage or cars.

Biodiesel

Biodiesel is the most common biofuel in Europe. It is produced from oils or fats using transesterification and is a liquid similar in composition to fossil/mineral diesel. Chemically, it consists mostly of fatty acid methyl (or

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ethyl) esters (FAMEs). Feedstocks for biodiesel include animal fats, vegetable oils, soy, rapeseed, jatropha, mahua, mustard, flax, sunflower, palm oil, hemp, field pennycress, pongamia pinnata and algae. Pure biodiesel (B100) is the lowest emission diesel fuel. Although liquefied petroleum gas and hydrogen have cleaner combustion, they are used to fuel much less efficient petrol engines and are not as widely available.

Biodiesel can be used in any diesel engine when mixed with mineral diesel. In some countries manufacturers cover their diesel engines under warranty for B100 use, although Volkswagen of Germany, for example, asks drivers to check by telephone with the VW environmental services department before switching to B100. B100 may become more viscous at lower temperatures, depending on the feedstock used. In most cases, biodiesel is compatible with diesel engines from 1994 onwards, which use 'Viton' (by DuPont) synthetic rubber in their mechanical fuel injection systems.

Electronically controlled 'common rail' and 'unit injector' type systems from the late 1990s onwards may only use biodiesel blended with conventional diesel fuel. These engines have finely metered and atomized multi-stage injection systems that are very sensitive to the viscosity of the fuel. Many current generation diesel engines are made so that they can run on B100 without altering the engine itself, although this depends on the fuel rail design. Since biodiesel is an effective solvent and cleans residues deposited by mineral diesel, engine filters may need to be replaced more often, as the biofuel dissolves old deposits in the fuel tank and pipes. It also effectively cleans the engine combustion chamber of carbon deposits, helping to maintain efficiency. In many European countries, a 5% biodiesel blend is widely used and is available at thousands of gas stations. Biodiesel is also an oxygenated fuel, meaning that it contains a reduced amount of carbon and higher hydrogen and oxygen content than fossil diesel. This improves the combustion of fossil diesel and reduces the particulate emissions from un-burnt carbon.

Biodiesel is also safe to handle and transport because it is as biodegradable as sugar, 10 times less toxic than table salt, and has a high flash point of about 300 F (148 C) compared to petroleum diesel fuel, which has a flash point of 125 F (52 C).

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Vegetable oil

Straight unmodified edible vegetable oil is generally not used as fuel, but lower quality oil can be used for this purpose. Used vegetable oil is increasingly being processed into biodiesel, or (more rarely) cleaned of water and particulates and used as a fuel.

Also here, as with 100% biodiesel (B100), to ensure that the fuel injectors atomize the vegetable oil in the correct pattern for efficient combustion, vegetable oil fuel must be heated to reduce its viscosity to that of diesel, either by electric coils or heat exchangers. This is easier in warm or temperate climates. Big corporations like MAN B&W Diesel, Wärtsilä, and Deutz AG as well as a number of smaller companies such as Elsbett offer engines that are compatible with straight vegetable oil, without the need for after-market modifications.

Vegetable oil can also be used in many older diesel engines that do not use common rail or unit injection electronic diesel injection systems. Due to the design of the combustion chambers in indirect injection engines, these are the best engines for use with vegetable oil. This system allows the relatively larger oil molecules more time to burn. Some older engines, especially Mercedes are driven experimentally by enthusiasts without any conversion, a handful of drivers have experienced limited success with earlier pre-"Pumpe Duse" VW TDI engines and other similar engines with direct injection. Several companies like Elsbett or Wolf have developed professional conversion kits and successfully installed hundreds of them over the last decades.

Oils and fats can be hydrogenated to give a diesel substitute. The resulting product is a straight chain hydrocarbon, high in cetane, low in aromatics and sulfur and does not contain oxygen. Hydrogenated oils can be blended with diesel in all proportions Hydrogenated oils have several advantages over biodiesel, including good performance at low temperatures, no storage stability problems and no susceptibility to microbial attack.[18]

Bioethers

Bio ethers (also referred to as fuel ethers or oxygenated fuels) are cost-effective compounds that act as octane rating enhancers. They also enhance engine performance, whilst significantly reducing engine wear and

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toxic exhaust emissions. Greatly reducing the amount of ground-level ozone, they contribute to the quality of the air we breathe.

Biogas

Biogas is methane produced by the process of anaerobic digestion of organic material by anaerobes. It can be produced either from biodegradable waste materials or by the use of energy crops fed into anaerobic digesters to supplement gas yields. The solid byproduct, digestate, can be used as a biofuel or a fertilizer.

Biogas can be recovered from mechanical biological treatment waste processing systems.

Note: Landfill gas is a less clean form of biogas which is produced in landfills through naturally occurring anaerobic digestion. If it escapes into the atmosphere it is a potential greenhouse gas.

Farmers can produce biogas from manure from their cows by getting a anaerobic digester (AD).

Syngas

Syngas, a mixture of carbon monoxide and hydrogen, is produced by partial combustion of biomass, that is, combustion with an amount of oxygen that is not sufficient to convert the biomass completely to carbon dioxide and water. Before partial combustion the biomass is dried, and sometimes pyrolysed. The resulting gas mixture, syngas, is more efficient than direct combustion of the original biofuel; more of the energy contained in the fuel is extracted.

Syngas may be burned directly in internal combustion engines or turbines. The wood gas generator is a wood-fueled gasification reactor mounted on an internal combustion engine.

Syngas can be used to produce methanol, DME and hydrogen, or converted via the Fischer-Tropsch process to produce a diesel substitute, or a mixture of alcohols that can be blended into gasoline. Gasification normally relies on temperatures >700°C.

Lower temperature gasification is desirable when co-producing biochar but results in a Syngas polluted with tar.

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Solid biofuels

When raw biomass is already in a suitable form (such as firewood), it can burn directly in a stove or furnace to provide heat or raise steam. When raw biomass is in an inconvenient form (such as sawdust, wood chips, grass, urban waste wood, agricultural residues), the typical process is to densify the biomass. This process includes grinding the raw biomass to an appropriate particulate size (known as hogfuel), which depending on the densification type can be from 1 to 3 cm (1 in), which is then concentrated into a fuel product. The current types of processes are wood pellet, cube, or puck. The pellet process is most common in Europe and is typically a pure wood product. The other types of densification are larger in size compared to a pellet and are compatible with a broad range of input feedstocks. The resulting densified fuel is easier to transport and feed into thermal generation systems such as boilers.

A problem with the combustion of raw biomass is that it emits considerable amounts of pollutants such as particulates and PAHs (polycyclic aromatic hydrocarbons). Even modern pellet boilers generate much more pollutants than oil or natural gas boilers. Pellets made from agricultural residues are usually worse than wood pellets, producing much larger emissions of dioxins and chlorophenols.

Notwithstanding the above noted study, numerous studies have shown that biomass fuels have significantly less impact on the environment than fossil based fuels. Of note is the U.S. Department of Energy Laboratory, Operated by Midwest Research Institute Biomass Power and Conventional Fossil Systems with and without CO2 Sequestration – Comparing the Energy Balance, Greenhouse Gas Emissions and Economics Study. Power generation emits significant amounts of greenhouse gases (GHGs), mainly carbon dioxide (CO2). Sequestering CO2 from the power plant flue gas can significantly reduce the GHGs from the power plant itself, but this is not the total picture. CO2 capture and sequestration consumes additional energy, thus lowering the plant's fuel-to-electricity efficiency. To compensate for this, more fossil fuel must be procured and consumed to make up for lost capacity.

Taking this into consideration, the global warming potential (GWP), which is a combination of CO2, methane (CH4), and nitrous oxide (N2O) emissions, and energy balance of the system need to be examined using a

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life cycle assessment. This takes into account the upstream processes which remain constant after CO2 sequestration as well as the steps required for additional power generation. firing biomass instead of coal led to a 148% reduction in GWP.

A derivative of solid biofuel is biochar, which is produced by biomass pyrolysis. Bio-char made from agricultural waste can substitute for wood charcoal. As wood stock becomes scarce this alternative is gaining ground. In eastern Democratic Republic of Congo, for example, biomass briquettes are being marketed as an alternative to charcoal in order to protect Virunga National Park from deforestation associated with charcoal production.

Second generation biofuels

Supporters of biofuels claim that a more viable solution is to increase political and industrial support for, and rapidity of, second-generation biofuel implementation from non-food crops. These include waste biomass, the stalks of wheat, corn, wood, and special-energy-or-biomass crops (e.g. Miscanthus). Some second generation (2G) biofuels use biomass to liquid technology, including cellulosic biofuels. Many second generation biofuels are under development such as biohydrogen, biomethanol, DMF, BioDME, Fischer-Tropsch diesel, biohydrogen diesel, mixed alcohols and wood diesel.

Cellulosic ethanol production uses non-food crops or inedible waste products and does not divert food away from the animal or human food chain. Lignocellulose is the "woody" structural material of plants. This feedstock is abundant and diverse, and in some cases (like citrus peels or sawdust) it is in itself a significant disposal problem.

Producing ethanol from cellulose is a difficult technical problem to solve. In nature, ruminant livestock (like cattle) eat grass and then use slow enzymatic digestive processes to break it into glucose (sugar). In cellulosic ethanol laboratories, various experimental processes are being developed to do the same thing, and then the sugars released can be fermented to make ethanol fuel. In 2009 scientists reported developing, using "synthetic biology", "15 new highly stable fungal enzyme catalysts that efficiently break down cellulose into sugars at high temperatures", adding to the 10 previously known. The use of high temperatures, has been identified as an important factor in improving the overall economic feasibility of the biofuel industry and the identification of enzymes that are stable and can operate efficiently at extreme temperatures is an area of active research. In

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addition, research conducted at TU Delft by Jack Pronk has shown that elephant yeast, when slightly modified can also create ethanol from non-edible ground sources (e.g. straw).

The recent discovery of the fungus Gliocladium roseum points toward the production of so-called myco-diesel from cellulose. This organism was recently discovered in the rainforests of northern Patagonia and has the unique capability of converting cellulose into medium length hydrocarbons typically found in diesel fuel. Scientists also work on experimental recombinant DNA genetic engineering organisms that could increase biofuel potential.

Scientists working in New Zealand have developed a technology to use industrial waste gases from steel mills as a feedstock for a microbial fermentation process to produce ethanol.

Second, third, and fourth generation biofuels are also called advanced biofuels.

Third generation biofuels

Algae fuel, also called oilgae or third generation biofuel, is a biofuel from algae. Algae are low-input, high-yield feedstocks to produce biofuels. Based on laboratory experiments, it is claimed that algae can produce up to 30 times more energy per acre than land crops such as soybeans, but these yields have yet to be produced commercially. With the higher prices of fossil fuels (petroleum), there is much interest in algaculture (farming algae). One advantage of many biofuels over most other fuel types is that they are biodegradable, and so relatively harmless to the environment if spilled. Algae fuel still has its difficulties though, for instance to produce algae fuels it must be mixed uniformly, which, if done by agitation, could affect biomass growth.

The United States Department of Energy estimates that if algae fuel replaced all the petroleum fuel in the United States, it would require only 15,000 square miles (38,849 square kilometers), which is roughly the size of Maryland, or less than one seventh the amount of land devoted to corn in 2000.

Algae, such as Botryococcus braunii and Chlorella vulgaris are relatively easy to grow, but the algal oil is hard to extract. There are several

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approaches, some of which work better than others. Macroalgae (seaweed) also have a great potential for bioethanol and biogas production.

Ethanol from living algae

Most biofuel production comes from harvesting organic matter and then converting it to fuel but an alternative approach relies on the fact that some algae naturally produce ethanol and this can be collected without killing the algae. The ethanol evaporates and then can be condensed and collected. The company Algenol is trying to commercialize this process.

Fourth generation biofuels

A number of companies are pursuing advanced "bio-chemical" and "thermo-chemical" processes that produce "drop in" fuels like "green gasoline," "green diesel," and "green aviation fuel." While there is no one established definition of "fourth-generation biofuels," some have referred to it as the biofuels created from processes other than first generation ethanol and biodiesel, second generation cellulosic ethanol, and third generation algae biofuel. Some fourth generation technology pathways include: pyrolysis, gasification, upgrading, solar-to-fuel, and genetic manipulation of organisms to secrete hydrocarbons.

GreenFuel Technologies Corporation developed a patented bioreactor system that uses nontoxic photosynthetic algae to take in smokestacks flue gases and produce biofuels such as biodiesel, biogas and a dry fuel comparable to coal.

With thermal depolymerization of biological waste one can extract methane and other oils similar to petroleum.

Hydrocarbon plants or petroleum plants are plants which produce terpenoids as secondary metabolites that can be converted to gasoline-like fuels. Latex producing members of the Euphorbiaceae such as Euphorbia lathyris and E. tirucalli and members of Apocynaceae have been studied for their potential energy uses.

Green fuels

However, if biocatalytic cracking and traditional fractional distillation are used to process properly prepared algal biomass i.e. biocrude, then as a

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result we receive the following distillates: jet fuel, gasoline, diesel, etc.. Hence, we may call them third generation or green fuels.

BioleachingBiomining or bioleaching is the microbial (mainly bacterial) process of

mineral extraction through leaching from low-grade ores (copper, gold, uranium, etc.)

Bioleaching can involve numerous ferrous iron and sulfur oxidizing bacteria, including Acidithiobacillus ferrooxidans and Acidithiobacillus (formerly known as Thiobacillus). As a general principle, Fe3+ ions are used as an oxidize the ore. This step is entirely independent of microbes. The role of the bacteria is the further oxidation of the ore, but more importantly also the regeneration of the chemical oxidant Fe3+ from Fe<sup2+/>. For example, bacteria catalyse the breakdown of the mineral pyrite (FeS2) by oxidising the sulfur and metal (in this case ferrous iron, Fe2+) using oxygen. This yields soluble products which can be further purified and refined to yield the desired metal.

Pyrite leaching (FeS2): In the first step, disulfide is spontaneously oxidized to thiosulfate by ferric iron (Fe3+), which in turn is reduced to give ferrous iron (Fe2+):

(1)      spontaneousThe ferrous iron is then oxidized by bacteria using oxygen:

(2)      (iron oxidizers)Thiosulfate is also oxidized by bacteria to give sulfate:

(3)      (sulfur oxidizers)

The ferric iron produced in reaction (2) oxidized more sulfide as in reaction (1), closing the cycle and given the net reaction:

(4)  

The net products of the reaction are soluble ferrous sulfate and sulfuric acid.The microbial oxidation process occurs at the cell membrane of the bacteria. The electrons pass into the cells and are used in biochemical processes to produce energy for the bacteria while reducing oxygen to

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water. The critical reaction is the oxidation of sulfide by ferric iron. The main role of the bacterial step is the regeneration of this reactant.

The process for copper is very similar, but the efficiency and kinetics depend on the copper mineralogy. The most efficient minerals are supergene minerals such as chalcocite, Cu2S and Covellite, CuS. The main copper mineral chalcopyrite (CuFeS2) is not leached very efficiently, which is why the dominant copper producing technology remains flotation followed by smelting and refining. The leaching of CuFeS2 follows the two stages of being dissolved and then further oxidised, with Cu2+ ions being left in solution.

Chalcopyrite leaching:

(1)      spontaneous

(2)      (iron oxidizers)

(3)      (sulfur oxidizers)net reaction:

(4)  

In general, sulfides are first oxidized to elemental sulfur, whereas disulfides are oxidized to give thiosulfate, and the processes above can be applied to other sulfidic ores. Bioleaching of non-sulfidic ores such as pitchblende also uses ferric iron as an oxidant (e.g. UO2 + 2 Fe3+ ==> UO2

2+ + 2 Fe2+). In this case the sole purpose of the bacterial step is the regeneration of Fe3+. Sulfidic iron ores can be added to speed up the process and provide a source of iron.

Further processing

The dissolved copper (Cu2+) ions are removed from the solution by ligand exchange solvent extraction which leaves other ions in the solution. The copper is removed by bonding to a ligand, which is a large molecule consisting of a number of smaller groups, each possessing a lone electron pair. The ligand-copper complex is extrcted from the solution using an organic solvent such as kerosene:

Cu2+(aq) + 2LH(organic) → CuL2(organic) + 2H+

(aq)

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The ligand donates electrons to the copper, producing a complex - a central metal atom (copper) bonded to the ligand. Because this complex has no charge, it is no longer attracted to polar water molecules and dissolves in the kerosene, which is then easily separated from the solution. Because the initial reaction is reversible, it is determined by pH. Adding concentrated acid reverses the equation, and the copper ions go back into an aqueous solution.Then the copper is passed through an electro-winning process to increase its purity: an electric current is passed through the resulting solution of copper ions. Because copper ions have a 2+ charge, they are attracted to the negative cathodes and collect there.The copper can also be concentrated and separated by displacing the copper with Fe from scrap iron:Cu2+

(aq) + Fe(s) → Cu(s) + Fe2+(aq)

The electrons lost by the iron are taken up by the copper. Copper is the oxidising agent (it accepts electrons), and iron is the reducing agent (it loses electrons).

Traces of precious metals such as gold may be left in the original solution. Treating the mixture with sodium cyanide in the presence of free oxygen dissolves the gold. The gold is removed from the solution by adsorbing (taking it up on the surface) to charcoal.

Bioleaching with fungi

Several species of fungi can be used for bioleaching. Fungi can be grown on many different substrates, such as electronic scrap, catalytic converters, and fly ash from municipal waste incineration. Experiments have shown that two fungal strains (Aspergillus niger, Penicillium simplicissimum) were able to mobilize Cu and Sn by 65%, and Al, Ni, Pb, and Zn by more than 95%.Aspergillus niger can produce some organic acids such as citric acid This form of leaching does not rely on microbial oxidation of metal, but rather uses microbial metabolism as source of acids which directly dissolve the metal.

Compared with other extraction techniques

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Extractions involve many expensive steps such as roasting and smelting, which require sufficient concentrations of elements in ores and are environmentally unfriendly. Low concentrations are not a problem for bacteria because they simply ignore the waste which surrounds the metals, attaining extraction yields of over 90% in some cases. These microorganisms actually gain energy by breaking down minerals into their constituent elements. The company simply collects the ions out of the solution after the bacteria have finished. Some advantages associated with bioleaching are:

economical: bioleaching is generally simpler and therefore cheaper to operate and maintain than traditional processes, since fewer specialists are needed to operate complex chemical plants.

environmental: The process is more environmentally friendly than traditional extraction methods. For the company this can translate into profit, since the necessary limiting of sulfur dioxide emissions during smelting is expensive. Less landscape damage occurs, since the bacteria involved grow naturally, and the mine and surrounding area can be left relatively untouched. As the bacteria breed in the conditions of the mine, they are easily cultivated and recycled.

Some disadvantages associated with bioleaching are:

economical: the bacterial leaching process is very slow compared to smelting. This brings in less profit as well as introducing a significant delay in cash flow for new plants.

environmental: Toxic chemicals are sometimes produced in the process. Sulfuric acid and H+ ions which have been formed can leak into the ground and surface water turning it acidic, causing environmental damage. Heavy ions such as iron, zinc, and arsenic leak during acid mine drainage. When the pH of this solution rises, as a result of dilution by fresh water, these ions precipitate, forming "Yellow Boy" pollution. For these reasons, a setup of bioleaching must be carefully planned, since the process can lead to a biosafety failure.

Currently it is more economical to smelt copper ore rather than to use bioleaching, since the concentration of copper in its ore is generally quite high. The profit obtained from the speed and yield of smelting justifies its cost. However, the concentration of gold in its ore is generally very low. The

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lower cost of bacterial leaching in this case outweighs the time it takes to extract the metal.

Enzyme EngineeringThis means changing the primary structure of the

existing-proteins/enzymes by genetic engineering. The protein/enzyme can be modified into a more efficient prototype. By engineering proteins, immunotoxins can be produced which can destroy specific cell types.

Production of enzymesMany moulds synthesise and excrete large quantities of enzymes into

the surrounding medium. Enzymes are proteins; they are denatured by heat and extracted or precipitated by chemical solvents like ethanol and by inorganic salts like ammonium sulphate. Coenzymes are also proteins combined with low molecular mass organics like vitamin B. It is industrially applicable and economically feasible to produce, concentrate, extract and purify enzymes from cultures of moulds such as Aspergillus, Penicillium, Mucor and Rhizopus. Mould enzymes such as amylase, invertase, protease, and pectinase are useful in the processing or refining of a variety of materials. Amylases hydrolyse starch to dextrin and sugars. They are used in preparing sizes and adhesives, desizing textile, clarifying fruit juices, manufacturing pharmaceuticals and other purposes. Invertase hydrolyses sucrose to form glucose and fructose (invert sugar). It is widely used in candy making and the production of non-crystallizable syrup from sucrose, which is partly hydrolysed by this enzyme. The proteolytic enzymes such as protease are used for bating in leather processing to obtain fine texture. Protease is also used in the manufacture of liquid glue, degumming of silks and clarification of beer protein. It is used in laundry detergents and as an adjunct with soaps. Pectinase is used in the clarification of fruit juice and to hydrolyse pectins in the retting of flax for the manufacture of linen. Apoenzyme is the protein portion of the enzyme, which is inactive. The reaction between low molecular mass coenzymes and apoenzyme gives active holoenzyme:

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E. Biodiversity ConservationBiodiversity can be measured at the gene level, the species level or at

the ecosystems level. The conservation of biodiversity is vital for our future survival and quality of life. Due to human interference (increased use of agricultural land, environmental pollution due to industrialization and mining) hundreds and thousand of species have been lost and thousands more are on the verge of extinction. Thus biodiversity conservation is becoming one of the priority areas of human activities. There is an urgent need for biodiversity conservation throughout the world.

Gene Bank and Plant ConservationTo feed the ever-growing human population, agriculture has been

extended to large areas and as a result, most of the cropping areas are occupied by a few high yielding crops (monoculture). Consequently, our natural resources are rapidly disappearing and the genetic diversity of many species is either decreasing or lost. Therefore, to save the valuable and threatened species, germplasm conservation is essential. Gene banks help preserve genetic material, be it plant or animal. In plants, this could be by freezing cuts from the plant, or stocking the seeds. In animals, this is the freezing of sperm and eggs in zoological freezers until

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further need. With corals, fragments are taken which are stored in water tanks under controlled conditions.

In plants, it is possible to unfreeze the material and propagate it, however, in animals; a living female is required for artificial insemination. While it is often difficult to utilize frozen animal sperm and eggs, there are many examples of it being done successfully.In an effort to conserve agricultural biodiversity, gene banks are used to store and conserve the plant genetic resources of major crop plants and their crop wild relatives. There are many gene banks all over the world, with the Svalbard Global Seed Vault being probably the most famous one.

The gene bank has undertaken the challenge of conserving the gene pool of many species. During the last two decades, many regional and international genetic resource centers (GRCs) have been set up in different countries. For example, the International rice Research Institute (IRRI), Manila has collected about 25,000 varieties of rice germplasm. Similarly, at the Maize and Wheat Improvement Center (CIMMYT), Mexico, more than 12,000 varieties have been conserved.

Svalbard Global Seed VaultThe Svalbard Global Seed Vault is a secure seedbank located on the

Norwegian island of Spitsbergen near the town of Longyearbyen in the remote Arctic Svalbard archipelago, about 1,300 kilometres (810 mi) from the North Pole. The facility preserves a wide variety of plant seeds in an underground cavern. The seeds are duplicate samples, or "spare" copies, of seeds held in genebanks worldwide. The seed vault will provide insurance against the loss of seeds in genebanks, as well as a refuge for seeds in the case of large scale regional or global crises.

F. EvolutionRecombinant DNA (rDNA) is a form of artificial DNA that is created by

combining two or more sequences that would not normally occur together through the process of gene splicing. In terms of genetic modification, it is created through the introduction of relevant DNA into an existing organismal DNA, such as the plasmids of bacteria, to code for or alter different traits for a specific purpose, such as antibiotic resistance. It differs from genetic recombination in that it does not occur through natural

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processes within the cell, but is engineered. A recombinant protein is a protein that is derived from recombinant DNA.

Recombinant DNA technology is of great service in bridging several missing links in the evolution. This has been done by amplifying the DNA (by Polymerase Chain Reaction or PCR) from the archaeological samples of extinct animals.

The polymerase chain reaction (PCR) is a scientific technique in molecular biology to amplify a single or a few copies of a piece of DNA across several orders of magnitude, generating thousands to millions of copies of a particular DNA sequence.

G. Environmental Biotechnology

Biodegradation of Oil SpillsOil spills and automobile exhaust are major sources of environmental

pollution in recent times, particularly in industrial areas. A strain of Pseudomonas putida has been used to treat environmental pollution. Many other efficient strains are in the developmental stages.

Petroleum oil is toxic for most life forms and episodic and chronic pollution of the environment by oil causes major ecological perturbations. Marine environments are especially vulnerable since oil spills of coastal regions and the open sea are poorly containable and mitigation is difficult. In addition to pollution through human activities, millions of tons of petroleum enter the marine environment every year from natural seepages. Despite its toxicity, a considerable fraction of petroleum oil entering marine systems is eliminated by the hydrocarbon-degrading activities of microbial communities, in particular by a remarkable recently discovered group of specialists, the so-called hydrocarbonoclastic bacteria (HCB). Alcanivorax borkumensis, a paradigm of HCB and probably the most important global oil degrader, was the first to be subjected to a functional genomic analysis. This analysis has yielded important new insights into its capacity for (i) n-alkane degradation including metabolism, biosurfactant production and biofilm formation, (ii) scavenging of nutrients and cofactors in the oligotrophic marine environment, as well as (iii) coping with various habitat-specific stresses. The understanding thereby gained constitutes a significant advance in efforts towards the design of new knowledge-based strategies for the mitigation of ecological damage caused by oil pollution of marine

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habitats. HCB also have potential biotechnological applications in the areas of bioplastics and biocatalysis.

Biological Control: An alternative to chemical control of plant diseases and pests

The biocontrol of diseases and insect pests, by using viruses, bacteria, amoebae, fungi, and other microorganisms is environmentally friendly and circumvents the use of pesticides, which cause environmental pollution and pose health hazards.

IV. ISSUES PERTAINING TO BIOTECH PRODUCTS

A. Intellectual Property Rights and Farmers’ Rights

INTERNATIONAL INTELLECTUAL PROPERTY FOR BIOTECHNOLOGY

Introduction

International intellectual property issues are becoming increasingly important as biotechnology is used and sold worldwide. This is because intellectual property rights are inextricably linked to the right to exclude others from use. The right to exclude can provide a competitive advantage or a barrier to entry into a commercial market. Several types of intellectual property provide some right to exclude others from an invention in the area of biotechnology—trade secrets, patents, and plant variety rights. Trade secrets enable their holder to prevent others from wrongfully appropriating valuable information for a potentially infinite time period. But they do not protect against independent invention, and they terminate once the information becomes public. A patent provides a right to exclude others, including independent inventors, from using the patented invention without consent, and only for a limited time. Plant variety rights function similarly to patents with respect to the ability to exclude others, but they are only available for plant ‘‘varieties.’’

Types of Protection

Trade Secrets. A trade secret typically consists of any information that is not generally known to others in the same business; it provides a

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competitive advantage to its owner. This is the easiest type of intellectual property to acquire, but it also provides the weakest protection. Unlike other types of intellectual property protection, formal procedural requirements are usually unnecessary to ‘‘obtain’’ a trade secret. Rather, the inventor of a trade secret merely needs to keep the information reasonably secret. The trade secret lasts as long as the information remains secret. However, once the information becomes publicly known, the trade secret ceases to exist. One way this can happen is if the information is independently developed and patented by another. Under this scenario, a trade secret would be exterminated because a patent on the same information would reveal the information to the public (since patents are public documents). Moreover, under this scenario, the patent owner could preclude the former trade secret owner from using the now patented invention because patent owners generally have rights to exclude all others from the patented invention. Although some countries soften this approach by providing those who used an invention prior to its patenting by another (prior users) with the right of continued use, there is no uniformity in such protection; for example, while some European countries allow a limited right, no analogous protection exists for biotechnology trade secrets in the United States.

Even without the complications of a superceding patent, trade secret protection offers minimal protection. To begin with trade secret protection does not allow exclusion of those who independently invent the identical ‘‘trade secret’’ therefore, two or more individuals or corporations could theoretically be using the same trade secret without infringing on each other’s rights if they all did so independently. A trade secret does not confer any affirmative rights except as to those who misappropriate the information (e.g., an employee who leaves with confidential information). Even then, the ‘‘protection’’ provided is usually inadequate because monetary compensation for the trade secret misappropriation cannot restore information to trade secrecy status if it has been disclosed to the public.

Patents. Patents provide inventors a reward or incentive for publicly disclosing an invention, by providing the inventor with the right to exclude others from the patented invention for a limited term. The exclusivity provided by a patent is considered critical to stimulate ideas and lead to further advances. The requirements established for patentability are aimed at securing the goal of promoting innovation. Although the requirements vary somewhat between various countries, the typical requirements for

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obtaining a patent are that (1) the invention constitute patentable subject matter, namely constitute the type of subject matter that country wants to encourage innovation in, and (2) that the invention, as disclosed in a patent application, satisfies technical patentability requirements. The scope of patentable subject matter may include both products and processes in all areas of technology. Technical patentability requirements typically require that the invention be at least ‘‘new,’’ ‘‘useful’’ (or have ‘‘industrial application’’), ‘‘nonobvious’’ (or have an ‘‘inventive step’’), and fully disclosed in a written document such that someone who was similarly technically competent could reproduce the invention. These requirements are intended to define inventive activity deserving of a patent. A national patent office typically examines patent applications to determine whether the patentability requirements are met. Additionally, in some countries, third parties are allowed to oppose the issuance of a patent or petition to revoke an existing patent for failing to meet the technical requirements.

Patents are generally considered the preferable type of intellectual property to protect biotechnology because they provide the most exclusive rights. Unlike trade secrets, patents protect against independent invention because the owner of a patent can exclude all others from using the patented invention. In addition, because a patent can entitle its owner to exclude others, including competitors, a patent or even a potential patent can justify the often high cost of research and development involved in biotechnology. The potential to exclude all others through patent protection is considered more valuable than attempting to maintain a trade secret indefinitely with no potential to affirmatively exclude others. Thus patents are the principal type of protection that is sought for biotechnology even though disclosure of the invention is required and patent protection is not a certainty.

Plant Variety Rights. A plant variety right also confers some exclusive rights. A plant variety right, which is also referred to as a ‘‘breeder’s right’’ (because the right is provided to the breeder of a plant variety), functions analogously to patent rights—a relatively exclusive right is provided to breeders of new plant varieties to further the development of agriculture. As with patents, plant variety rights are not automatic; rather, they must be applied for and examined to determine whether they meet the requisite technical requirements. The requirements for plant variety rights are intended to function similarly to those for patent rights in that both are intended to provide protection to subject matter that is truly innovative. Some of the technical requirements for plant breeder rights parallel those

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for patent rights—a variety must be ‘‘new,’’ ‘‘distinct,’’ ‘‘uniform,’’ and ‘‘stable.’’ These requirements are generally less onerous than those needed to meet patentability requirements. In addition, unlike patents, disclosure of the invention, or at least the method of making the invention, is not always required, which could be seen as an advantage. Although plant variety rights will be addressed in this article, the focus is on patent protection because patents provide coverage for more types of biotechnology and broadest protection.

International Conventions Impacting Biotechnology

1. European Patent ConventionThe Convention on the Grant of European Patents of 5 October 1973, commonly known as the European Patent Convention (EPC), is a multilateral treaty instituting the European Patent Organisation and providing an autonomous legal system according to which European patents are granted.2. European UnionThe European Union (EU) is an economic and political union of 25 member states which are located primarily in Europe.3. Agreement on Trade Related Aspects of Intellectual Property Rights (TRIPS)The Agreement on Trade Related Aspects of Intellectual Property Rights (TRIPS) is an international agreement administered by the World Trade Organization (WTO) that sets down minimum standards for many forms of intellectual property (IP) regulation as applied to nationals of other WTO Members.4. Budapest ConventionThe Budapest Treaty on the International Recognition of the Deposit of Microorganisms for the Purposes of Patent Procedure, or Budapest Treaty, is an international treaty signed in Budapest, Hungary, on April 28, 1977. It entered into force on August 9, 1980, and was later amended on September 26, 1980. The treaty is administered by the World Intellectual Property Organization (WIPO).5. International Union for the Protection of New Varieties of Plants (Union internationale pour la protection des obtentions végétales)The International Union for the Protection of New Varieties of Plants or UPOV is an intergovernmental organization with headquarters in Geneva, Switzerland. The current Secretary-General of UPOV is Francis Gurry.

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UPOV was established by the International Convention for the Protection of New Varieties of Plants. The Convention was adopted in Paris in 1961 and revised in 1972, 1978 and 1991. The objective of the Convention is the protection of new varieties of plants by an intellectual property right. By codifying intellectual property for plant breeders, UPOV aims to encourage the development of new varieties of plants for the benefit of society.

PHILIPPINE INTELLECTUAL PROPERTY FOR BIOTECHNOLOGY

Introduction

Technology transfer to developing countries is now more difficult because biotechnology has now become proprietary in nature and most developing countries do not have intellectual property (IP) laws or the capacity to implement them, if any. In the Philippines, there are two laws governing IP protection: the IP Code (1997), which contains a very broad definition of patents or patentable inventions or innovations (Section 21) and specifies that microorganisms, non-biological processes, and microbiological processes used for biotechnologies can be patented (Section 22); and the PVPA 2002, which allows plants with distinct, stable and uniform morphological characteristics to be patented. The law that provides IP protection for biotechnology products and processes is the IP Code but no mechanism to monitor infringement of IPR has yet been installed.

Biotechnology – Related International IPR Treaties to Which the Philippines is a Party

The Republic of the Philippines is a signatory to several international treaties and conventions on intellectual property rights, to wit:

Convention Establishing the World Intellectual Property Organization [since 1980] Budapest Treaty on the International Recognition of the Deposit of

Microorganisms for Purposes of Patent Procedure [since 1981] Agreement on Trade-Related Aspects of Intellectual Property Rights

[TRIPS Agreement]

RA 8293: Intellectual Property Code of the Philippines

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Important sections of the IP Code of the Philippines pertaining to biotechnology are stated below, which contains a very broad definition of patents or patentable inventions or innovations and specifies that microorganisms, non-biological processes, and microbiological processes used for biotechnologies can be patented.

Section 21. Patentable InventionsAny technical solution of a problem in any field of human activity which is new, involves an inventive step and is industrially applicable shall be Patentable. It may be, or may relate to, a product, or process, or an improvement of any of the foregoing. (Sec. 7, R.A. No. 165a)Section 22. Non-Patentable InventionsThe following shall be excluded from patent protection:22.1. Discoveries, scientific theories and mathematical methods;22.2. Schemes, rules and methods of performing mental acts, playing games or doing business, and programs for computers;22.3. Methods for treatment of the human or animal body by surgery or therapy and diagnostic methods practiced on the human or animal body. This provision shall not apply to products and composition for use in any of these methods;22.4. Plant varieties or animal breeds or essentially biological process for the production of plants or animals. This provision shall not apply to micro-organisms and non-biological and microbiological processes.Provisions under this subsection shall not preclude Congress to consider the enactment of a law providing sui generis protection of plant varieties and animal breeds and a system of community intellectual rights protection:22.5. Aesthetic creations; and22.6. Anything which is contrary to public order or morality. (Sec. 8,

R.A. No. 165a)

RA 9168: Philippine Plant Variety Protection Act of 2002

Congress of the PhilippinesTwelfth Congress

First Regular Session

REPUBLIC ACT NO. 9168June 7, 2002

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AN ACT TO PROVIDE PROTECTION TO NEW PLANT VARIETIES, ESTABLISHING A NATIONAL PLANT VARIETY PROTECTION BOARD

AND FOR OTHER PURPOSES

Be it enacted by the Senate and House of Representatives of the Philippines in Congress assembled:

Section 1. Short Title.This Act shall be known and cited as the "Philippine Plant Variety Protection Act of 2002"

Sec. 2. Statement of Policies.

a) The State recognizes that an effective intellectual property system in general and the development of new plant variety in particular is vital in attaining food security for the country. To this end, it shall protect and secure the exclusive rights of breeders with respect to their new plant variety particularly when beneficial to the people for such periods as provided for in this Act.

b) The use of intellectual property bears a socioeconomic function. To this end, the State shall promote the diffusion of technology and information for the promotion of national development and progress for the common good.

c) The State recognizes the indispensable role of the private sector, encourages the participation of private enterprises and provides incentives to needed investments in the development of new plant varieties.

d) The State recognizes that science and technology are essential for national development and promotes the adaptation of technology and knowledge from all sources for the national benefit. The State also recognizes the need to protect and secure the exclusive rights of scientists and other gifted citizens to their intellectual property and creations.

e) The State, while recognizing intellectual property rights in the field of agriculture, does so in a manner supportive of and not inconsistent with its obligation to maintain a healthful ecology in accord with the rhythm and harmony of nature.

TITLE II

Definitions

Sec. 3. Definitions. -

a) "Applicant" means the breeder who applies for the grant of a Certificate of Plant Variety Protection.

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b) "Board" means the National Plant Variety Protection Board created by this Act. It shall also refer to the National Seed Industry Council during the transition period from the effectivity of this Act up to the time the said Board has been organized and operating.

c) "breeder" means:

1. The person who bred, or discovered and developed a new plant variety; or

2. The person who is the employer of the aforementioned person or who has commissioned the work; or

3) The successors-in-interest of the foregoing persons as the case may be; or

4) The holder of the Certificate of Plant Variety Protection.

d) "Certificate of Plant Variety Protection" means the document issued by the Board pursuant to this Act for the protection of a new plant variety.

e) "Commission" means to engage the services of a person to develop new plant varieties in exchange for monetary or any material consideration.

f) "Harvested material" means any part of a plant with potential economic value or any product made directly therefrom in proper case.

g) "Holder" means a person who has been granted a Certificate of Plant Variety Protection or his successors-in-interest.

h) "Persons" includes natural persons and juridical persons.

i) "Plant" includes terrestrial and aquatic flora.

j) "Plant Variety Protection (PVP)" mans the rights of breeders over their new plant variety as defined in this Act.

k) "Propagating material" means any part of the plant that can be used to reproduce the protected variety.

l) "Regulations" means the rules and regulations promulgated by the Board for the purpose of implementing the provisions of this Act.

m) "Variety" means a plant grouping within a single botanical taxon of the lowest known rank, that without regard to whether the conditions for plant variety protection are fully met, can be defined by the expression of the

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characteristics resulting from a given genotype or combination of genotypes, distinguished from any other plant groupings by the expression of at least one (1) characteristics, and considered as a unit with regard to the suitability for being propagated unchanged. A variety may be represented by seed, transplants, plants, tubers, tissue culture plantlets, and other forms.

TITLE III

Conditions for the Grant of the Plant Variety Protection

Sec. 4. The Certificate of Plant Variety Protection shall be granted for varieties that are:

a) New;

b) Distinct;

c) Uniform; and

d) Stable.

Sec. 5. Newness. - A variety shall be deemed new if the propagating or harvested material of the variety has not been sold, offered for sale or otherwise disposed of to others, by or with the consent of the breeder, for purposes of exploitation of the variety;

a) In the Philippines for more than one (1) year before the date of filing of an application for plant variety protection; or

b) In other countries or territories in which the application has been filed, for more than four (4) years or, in the case of vines or tress, more than six (6) years before the date of filing of an application for Plant Variety Protection.

However, the requirement of novelty provided for in this Act shall not apply to varieties sold, offered for sale or disposed of to others for a period of five (5) years before the approval of this Act. Provided, That application for PVP shall be filed within one (1) year from the approval of this act.

Sec. 6. Distinctness. - A variety shall be deemed distinct if it is clearly distinguishable from any commonly known variety. The filing of an application for the granting of a plant variety protection or for the entering of a new variety in an official register of variety in the Philippines or in any country, shall render the said variety a matter of public knowledge from the date of the said application: Provided, That the application leads to the

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granting of a Certificate of Plant Variety Protection or the entering of the said other variety in the official register of variety as the case may be.

Sec. 7. Uniformity. - The variety shall be deemed uniform if, subject to the variation that may be expected from the particular features of its propagation, it is sufficiently uniform in its relevant characteristics.

Sec. 8. Stability. - The variety shall be deemed stable if its relevant characteristics remain unchanged after repeated propagation or, in the case of a particular cycle of propagation, at the end of each such cycle.

TITLE IV

Variety Denomination

Sec. 9. Variety Denomination. - The variety shall be designated by a denomination which shall be its generic description. In particular, it must be different from any denomination that designates an existing variety of the same plant species or closely related species.

Sec. 10. Right of Priority over Denomination. - The use of a denomination shall not be granted to a breeder if such denomination has already been registered to another breeder or is being used by a third party in relation to the sale or offering for sale of a particular variety prior to the filing date or priority date of an application for a Certificate of Plant Variety Protection. In case two (2) or more breeders/applicants apply for the registration of the same denomination, the breeder/applicant who has the earliest filing date or priority date shall have the right to register the same to the exclusion of the other applicant/breeder(s).

Sec. 11. Figures as Denomination. - The denomination must enable the variety to be identified. It may not consist solely of figures except when it is an established practice for designating such a variety.

Sec. 12. Misleading Denomination. - No denomination shall be accepted if it is liable to mislead or to cause confusion concerning the characteristic value or identity of the variety or identity of the breeder.

Sec. 13. Refusal of Denomination. - If the denomination does not satisfy these requirements, its registration shall be refused and the breeder shall be required to propose another denomination within a prescribed period. The denomination shall be registered together with the grant of the breeder's right.

Sec. 14. Denomination Used in an Application Previously Filed abroad. - An application filed in this country, the subject matter of which is

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the same as that of an application previously filed abroad, shall use the same denomination as the latter. However, if such denomination does not conform to the provisions of this Title, the applicant/breeder shall be required to submit a new denomination.

Sec. 15. Obligation to Use Denomination. - Any person, who offers for sale or markets in the Philippines, propagating material of a variety protected, shall be obliged to use the denomination of that variety, even after the expiration of the breeder's right therefor except when the rule of prior rights apply.

Sec. 16. Use of Marks. - When a protected variety is offered for sale or marketed, it may be associated with a trademark, trade name or other similar indication with a registered denomination. If such an indication is so associated, the denomination must nevertheless be easily recognizable.

TITLE V

Applicants to a Plant Variety Protection

Sec. 17. Entitlement. - Any breeder, with respect to the variety developed, may apply for a plant variety developed, may apply for a plant variety protection and obtain a Certificate of Plant Variety Protection upon compliance with the requirements of this Act.

Sec. 18. Co-ownership of the Right. - If two (2) or more persons contribute to the development of a new plant variety, all of them shall be named in the Certificate of Plant Variety Protection and shall be entitled to such rights as agreed upon in writing or in the absence thereof, the rights in proportion to their contribution in the development of plant variety.

Sec. 19. Employee-Employer relationship. - in case an employee develops a plant variety in the course of his employment as a result of the performance of his regular duty, the plant variety protection shall belong to the employer, unless there is a written stipulation to the contrary.

Sec. 20. First to File Rule. - If two (2) or more persons develop a new plant variety separately and independently of each other, the Certificate of Plant Variety Protection shall belong to the person who files the application first. In case two (2) or more persons file an application for the same plant variety, the right shall be granted to the person who has the earliest filing date or priority date.

Sec. 21. Priority Date. - Any application for a Certificate of Plant Variety Protection previously filed by a breeder in another country, which by treaty, convention or law affords similar privileges to Filipino citizens, shall be

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considered as filed locally as of the date of filing of the foreign application: Provided, That:

a) The local application expressly claims priority;

b) It is filed within twelve (12) months from the filing date of the earliest foreign application; and

c) The applicant submits, within six (6) months from the filing of the local application, authenticated copies of documents which constitute the foreign application, samples or other evidence showing that the variety which is being applied for protection is the same variety which has been applied for protection in a foreign country.

Sec. 22. Foreign Nationals. - For purposes of this Act, a person shall be considered a national of a foreign country if he is a citizen of such country according to its laws, a natural person residing therein, or is a legal entity whose office is registered in such foreign country.

Sec. 23. National Treatment. - Any application filed locally for a Certificate of Plant Variety Protection previously granted to a breeder in another country, which by treaty, convention or law affords similar privileges to Filipino citizens, shall be issued a Certificate of Plant Variety Protection upon payment of dues and compliance to all the provisions of this Act. This Act shall also apply to the nationals of foreign countries that are members of intergovernmental organizations or party to any multilateral agreement or convention concerning the granting of intellectual property protection to plant varieties.

TITLE VI

Examination of the Application and Issuance of PVP Certificate

Sec. 24. Contents of the Application. - An application for a Certificate of Plant Variety Protection shall be filed in the manner and on the conditions prescribed in the regulations, and shall include:

a) Name of the applicant/breeder;

b) Address of the applicant/breeder in the Philippines;

c) Name of resident agent and address in the Philippines;

d) The description of the variety bred, including particulars of its characteristics;

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e) The variety denomination;

f) Sample of propagating materials, which are the subject of the application; and

g) Any other particular required by the regulations.

Section 25. Rights of the Applicant to File the Application. - If the applicant is not the actual breeder, he shall indicate in his application the basis for his right to file the application.

Sec. 26. Contents of the Description and Order of Presentation. - The Board shall issue rules and regulations stipulating the contents of the description and the order of presentation.

Sec. 27. Other Information Required. - The applicant shall be required by the Board to furnish information regarding any application filed by him in other countries including all pertinent documents relating thereto. If the applicant has successfully claimed priority according to this Act, he shall be given a period of two (2) years from the priority date to comply with the requirements of this Sec. .

Sec. 28. Manner of Conducting Tests. - The Board may carry out the necessary tests, cause the conduct of tests, or consider the results of other tests or trials that have already been done. For this purpose, the Board shall require the applicant to furnish all the necessary information, documents or materials within a period of time prescribed in the regulations.

Sec. 29. Filing Date. - For purposes of according a filing date, the Board shall consider, as a minimum requirement, all of the above enumerated items in Sec. 24 hereof.

Sec. 30. Publication of the Application. - After the Board has accorded a filing date, the application shall be published within sixty (60) days at the expense of the applicant in the Plant Variety Gazette hereunder described in Sec. 73.

Prior to such publication, the application and all related documents shall not be made available to the public without the written consent of the applicant.

After publication of the application, any person may inspect the application documents in a manner to be prescribed by the Board.

Sec. 31. Opposition to the Grant of Plant Variety Protection. - Any person who believes that the applicant is not entitled to the grant of the

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Certificate of Plant Variety Protection may file an opposition thereto within the period prescribed by the Board from the date of its publication and before the issuance of the Certificate of Plant Variety Protection.

Opposition to the application may be made on the following grounds:

a) that the person opposing the application is entitled to the breeder's right as against the applicant;

b) that the variety is not registrable under this Act.

If the opposition is based on the conditions of Plant Variety Protection, such opposition shall be considered together with the examination of the application.

Sec. 32. Issuance of the Certificate. - When the Board has tested and examined the variety, and/or considered the supporting materials and literature pertinent thereto, it shall issue a Certificate of Plant Variety Protection. A notice of such issuance shall be published in the manner to be prescribed in the regulations at the expense of the holder.

Sec. 33. Term of Protection. - For trees and vines, the period of protection shall be twenty-five (25) years from the date of the grant of the Certificate of Plant Variety Protection and twenty (20) years from the said date for all other types of plants, unless declared void ab initio or cancelled otherwise, as provided under Sec. 61 and 62, respectively of this Act.

Sec. 34. Annual Fees. - To maintain the validity of the Certificate of Plant Variety Protection, the holder shall pay an annual fee to be prescribed by the Board. Annual fees shall be paid starting from the fourth anniversary of the issuance of the certificate and every year thereafter with the first three (3) months of said years. The holder has the option to pay in advance this annual fee for a maximum of twenty (20) years.

The Certificate of Plant Variety Protection shall expire and cease to have force and effect upon the holder's failure to pay the annual fees within the prescribed period. A notice of such cancellation shall be published in the Plant Variety Gazette one (1) year after the term of protection has expired. Before such publication, any holder who fails to pay the annual fees may request for a reinstatement of his certificate: Provided, That he settles his unpaid accounts including surcharges to be determined by the Board.

Sec. 35. Notice of Rejection. - Whenever an application is rejected, the Board shall immediately inform the applicant on the grounds therefor, and when applicable, identify and provide the documents used as the basis for rejection.

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a) Reconsideration - Within two (2) months from the receipt of the rejection notice, the applicant may amend his application or traverse the finding of the Board. The Board, in turn, may reverse its initial finding or issue a final rejection within the same period.

b) Appeal from the Notice of Rejection - The decision applicant of the Board is final except for anomalous circumstances involving the Board in which case the may appeal it to the proper court.

TITLE VII

Rights of Holders

Sec. 36. Rights of Holders of Plant Variety Protection. - In respect of the propagating materials, holders of a Certificate of Plant Variety Protection shall have the right to authorize any of the following acts:

a) Production or reproduction;

b) Conditioning for the purpose of propagation;

c) Offering for sale;

d) Selling or other marketing;

e) Exporting;

f) Importing; and

g) Stocking for any purpose mentioned above.

Sec. 37. The holder may make his authorization subject to conditions and limitations.

Sec. 38. Acts in Respect of Harvested Materials. - Except for Sections 43 and 44 of this Title, the rights in the two (2) preceding sections shall also extend to the harvested materials which may be the entire plant or its other parts, if the production thereof resulted directly from the unauthorized use of the plant's propagating materials that are covered by this Act, unless the holder has had the reasonable opportunity to exercise his right in relation to the said propagating materials.

Sec. 39. Coverage of Protection.- The rights of holder under Sections 36 and 38 of this Act shall also apply in relation to:

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a) Varieties which are essentially derived from the protected variety, where the protected variety is not itself an essentially derived variety;

b) Varieties which are not clearly distinct from the protected variety; and

c) Varieties whose production requires the repeated use of the protected variety.

Sec. 40. Essentially Derived Varieties. - For the purpose of paragraph 39(a), a variety shall be deemed to be essentially derived from the initial variety when:

a) it is predominantly derived from the initial variety, or from a variety that is itself predominantly derived from the initial variety, while retaining the expression of the essential characteristics that result from the genotype or combination of genotypes of the initial variety;

b) It is clearly distinguishable from the initial variety; and

c) Except for the differences which result from the act of derivation, it conforms to the initial variety in the expression of the essential characteristics that result from the genotype or combination of genotypes of the initial variety.

Sec. 41. Manner of Developing Essentially Derived Varieties. - It shall also be understood that essentially derived varieties may be obtained through processes which may include, but not limited to, the selection of a natural or induced mutant, or of a somoclonal variant, the selection of a variant individual from plants of initial variety, backcrossing or transformation by genetic engineering. Genetic engineering shall be understood as the introduction of genes by laboratory techniques.

Sec. 42. Provisional Protection. - An applicant for a Certificate of Plant Variety Protection shall be entitled to equitable remuneration from any person who, during the period between the publication of the application for the certificate and the grant of that certificate, has carried out acts which, once the certificate is granted, required the holder's authorization as conferred in this Act: Provided, That the applicant shall initiate the legal action against the alleged infringer within two (2) years from the date of the granting of his Certificate of Plant Variety Protection.

Sec. 43. Exceptions to Plant Variety Protection. - The Certificate of Plant Variety Protection shall not extent to:

a) Acts done for noncommercial purposes;

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b)Acts done for experimental purposes;

c) Acts done for the purpose of breeding other varieties, except when Sections 39 and 40 apply; and

d) The traditional right of small farmers to save, use, exchange, share or sell their farm produce of a variety protected under this Act, except when a sale is for the purpose of reproduction under a commercial marketing agreement. The Board shall determine the condition under which this exception shall apply, taking into consideration the nature of the plant cultivated, grown or sown. This provision shall also extend to the exchange and sell of seeds among and between said small farmers: Provided, That the small farmers may exchange or sell seeds for reproduction and replanting in their own land.

Sec. 44. Exhaustion of Plant Variety Protection. - The Certificate of Plant Variety Protection shall not extend to acts concerning any material of the protected variety, or a variety covered by the provisions of Sections 39 and 40 hereof, which has been sold or otherwise marketed by the breeder or with his consent in the Philippines, or any material derived from the said material, unless it:

a) Involves further propagation of the variety in question; or

b) Involves the export of the variety, which enables the propagation of the variety, into a country that does not protect the variety of the plant genus or species to which the variety belongs, except where the exported material is for final consumption purposes.

Sec. 45. Rights of Attribution. - No Certificate of Plant Variety Protection shall be issued without naming the breeder(s) unless this right is protested in writing within one (1) year.

Sec. 46. Succession/Transmission. - The Certificate of Plant Variety Protection shall be considered as a property right and the transmission thereof shall be governed by the law on Property.

TITLE VIII

Infringement

Sec. 47. What Constitutes Infringement. - Except as otherwise provided in this Act, any person who without being entitled to do so, performs the following acts:

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a) Sell the novel variety, or offer it or expose it for sale, deliver it, ship it, consign it, exchange it, or solicit an offer to buy it, or any other transfer of title or possession of it; or

b) Import the novel variety into, or export it from, the Philippines; or

c) Sexually multiply the novel variety as a step in marketing (for growing purposes) the variety; or

d) Use the novel variety in producing (as distinguished from developing) a hybrid or different variety therefrom; or

e) Use seed which had been marked "unauthorized propagation prohibited" or "unauthorized seed multiplication prohibited" or progeny thereof to propagate the novel variety; or

f) Dispense the novel variety to another, in a form which can be propagated, without notice as to being a protected variety under which it was received; or

g) Fails to use a variety denomination the use of which is obligatory under Sec. 15; or

h) Perform any of the foregoing acts even in instances in which the novel variety is multiplied other than sexually, except in pursuance of a valid Philippine plant patent; or

i) Instigate or actively induce performance of any foregoing acts, may be sued by the holder, who may also avail of all such relief as are available in any proceeding involving infringements of other proprietary rights.

Sec. 48. Where to Commence Action. - Any holder may petition the proper regional trial court for infringement of his plant variety protection as defined in this Act.

Sec. 49. Presumption of Validity. - Certificate of Plant Variety Protection shall be presumed valid and the burden of proof of their invalidity shall rest on the party assailing them.

Sec. 50. Defenses Against Infringement Charges. - The following shall be valid defenses against infringement charges:

a) Non-infringement;

b) The plant variety does not possess at the time of its application criterion of novelty or distinctness;

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c) The alleged infringement was performed under a right adverse to it, prior to the notice of infringement; and/or

d) Other defenses that are made available under this Act.

Sec. 51. Notice. - No damages shall be awarded unless there is actual or constructive notice made upon the alleged infringer.

Sec. 52. Damages. - The court may award actual, moral, exemplary damages and attorney's fees according to a proven amount including a reasonable royalty for the use of the protected variety.

Sec. 53. Injunction. - The court may also enjoin the infringer(s) from further performing any act of infringement on the rights of the holder(s) as defined in this Act.

Sec. 54. Court to Order Confiscation of Infringing Materials. - Upon petition by the complainant, the court may order the confiscation of infringing materials, and:

a) Cause their distribution to charitable organization;

b) Cause the sale and provide the proceeds thereof to research organizations; or

c) Cause the return to the petitioner for further scientific use.

Sec. 55. Prescription. - No recovery of damages for any infringement case shall prosper when the cause of action has reached more than six (6) years from the time the alleged infringement case was committed.

Sec. 56. Criminal Penalty. - Any person who violates any of the rights of the holder provided for in this Act may also suffer the penalty of imprisonment of not less than three (3) years but not more than six (6) years and/or a fine of up to three (3) times the profit derived by virtue of the infringement but in no case should be less than One Hundred Thousand pesos (P100,000.00).

TITLE IX

Compulsory License

Sec. 57. Grounds for the Grant of Compulsory Licensing. - Any interested person may file a petition for compulsory license with the Board at any time after two (2) years from the grant of the Certificate of Plant

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Variety Protection under this Act when it is for the public interest to grant such compulsory license, and:

a) The reasonable requirements of the public for any part of the variety are not met; or

b) There is an overseas market for the sale of any part of the variety and the same are not met by the holder; or

c) The plant variety developed relates to or required in the production of medicine and/or any food preparation.

Sec. 58. Scope of Compulsory License. - The Board, upon petition by any interested party and upon proof of any of the foregoing grounds, may issue a decision:

a) Allowing the petitioner to produce in commercial quantity and distribute the variety protected or any part thereof; or

b) Requiring the holder to ensure the availability of the propagating materials of the variety protected; or

c) Requiring the petitioner to pay the holder with license fees in the form of reasonable royalties; and

d) Other such additional remedies at the Board may determine to be consistent with appropriate circumstances.

Sec. 59. Duration of the License. - A compulsory license shall be effective until the ground(s) for its issuance has been terminated as determined by the Board motu proprio or upon petition by party or parties and resolution by the Board.

Sec. 60. Procedure for Grant. - The Board shall provide in the rules and regulations the manner and procedure for granting compulsory licenses.

TITLE X

Cancellation and Nullity of Plant Variety Protection

Sec. 61. Grounds for Nullity. - The Certificate of Plant Variety Protection shall be declared void ab initio on any of the following grounds:

a) The grant of the Certificate of Plant Variety Protection was essentially based upon information and documents furnished by the applicant, wherein

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the conditions of distinctness, uniformity, stability, and newness were not complied with the time of the grant of the certificate; or

b) The Certificate of Plant Variety Protection was granted to a person who is not entitled to it, unless it is transferred to the person who is so entitled.

The effect of the declaration of nullity is that as if the Certificate of Plant Variety Protection was not issued.

Sec. 62. Grounds for Cancellation. - The Plant Variety Protection shall be cancelled on any of the following grounds:

a) The breeder does not provide the required information, documents, or materials necessary for verifying the maintenance of the variety; or

b) the breeder fails to pay the required fees to keep his or her rights in force or provides false information in his or her application; or

c) The breeder does not propose, within the time/period provided under the regulations, another suitable denomination if the denomination of the variety is cancelled after the grant of the Certificate of Plant Variety Protection; or

d) The conditions of uniformity and stability could not be maintained although these were present at the time of the issuance of the Certificate of Plant Variety Protection; or

e) The breeder entitled to the Certificate of Plant Variety Protection or the holder has relinquished his/her rights through a declaration in a public instrument filed with the registrar.

Sec. 63. Venue. - Any petition to cancel a Certificate of Plant Variety Protection shall originally be under the jurisdiction of the Board. Decisions of the Board may be appealable with the Court of Appeals within fifteen (15) days from the date of notice of the Board's final decision.

Sec. 64. Prescription. - The right to cancel a Certificate of Plant Variety Protection shall be instituted at any time within the term of protection of such right.

Sec. 65. Publication. - A notice of the filing of a petition to cancel a Certificate of Plant Variety Protection and the final order/decision on the same shall be published in the Plant Variety Gazette at the expense of the petitioner.

TITLE XI

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Institution

Sec. 66. National Plant Variety Protection Board. - There is hereby created National Plant Variety Protection Board which shall be composed of the following or their duly designated representatives:

a) The Secretary of the Department of Agriculture, as chairman;

b) The Secretary of the Department of Science and Technology, as co-chairman;

c) The Director-General of the Intellectual Property Office, as vice chairman;

d) The Director of the Bureau of Plant Industry;

e) The Director of the Institute of Plant Breeding of the University of the Philippines Los Baños;

f) The President of the Philippine Seed Industry Association;

g) A representative from a federation of small farmers' organizations to be nominated by the Secretary of Agriculture;

h) A representative from the scientific community to be nominated by the National Academy of Science and Technology; and

i) The Registrar (ex officio).

The members of the Board or their representatives must be Filipino citizens, have good moral character and should not have been convicted of a crime involving moral turpitude.

The Board shall perform the following functions:

a) Promulgate policy guidelines for the effective implementation of the provisions of this Act;

b) Have original and exclusive appellate jurisdiction over all acts of the Registrar;

c) Have original jurisdiction over petitions for compulsory licensing, nullity and cancellation of the Certificate of Plant Variety Protection;

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d) Institutionalize database of existing plant varieties, collected from foreign and local databases, within one (1) year from the effectivity of this act;

e) Call on resource persons to provide inputs that will be relevant in the performance of the tasks of the Board;

f) Organize the Registrar as it sees fit;

g) Approve capital expenditure and contracts of experts; and

h) Perform all other functions as may be required in the implementation of this Act.

Sec. 67. Rules and Regulations. - For the purpose of the preceding section, the Board with representatives from the Senate and House Committees on Agriculture, shall within six (6) months from the effectivity of this act, prescribe rules and regulations necessary for the implementation of its functions, or reorganize and create units therefore under its control and supervision.

Sec. 68. Fees. - The Board shall prescribe a schedule of fees to be charge against any applicant/breeder in the course of the application for a Certificate of Plant Variety Protection or in the maintenance therefor.

Sec. 69. Coordination and Cooperation with Other Institutions. - For the purpose of verifying certain facts such as but not limited to the requirements of stability, distinctness and uniformity, the Board may enter into agreements with other governmental or nongovernmental institutions both domestic and foreign under a set of conditions germane to its functions.

Further, the Board shall also designate appropriate state colleges and universities, bona fide research institutions, or appropriate nongovernmental research centers as testing centers for the distinctness, uniformity and stability of varieties.

Sec. 70. The PVP Fund. - There is hereby created PVP Fund, hereinafter referred to as the Fund, to be administered by the Board. All fees, fines and charges collected by the Board under this Act, shall be deposited in the Fund. The Board is hereby authorized to use and disburse the Fund. The Board is hereby authorized to use and disburse the Fund without need of approval by any government agency, and subject only to existing accounting and auditing rules and regulations for purposes of defraying the cost of operations in the delivery of its services to the public.

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Sec. 71. Gene Trust Fund. - There shall be an independent and separate trust fund established under this Act, to be administered by the Board, for the benefit of bona fide organizations or institutions managing and operating an accredited gene bank. An amount to be determined by the Board but not to exceed twenty percent (20%) of the fees and charges, shall be used for the purposes of the gene trust fund. The trust fund may also accept donations from national and international institutions and other organizations and individuals interested in strengthening genetic conservation.

Sec. 72. Farming Communities and Bona fide Farmers' Organizations. - Farming communities and bona fide farmers' organizations are encouraged to build an inventory of locally-bred varieties as an option to protect these resources from misappropriation and unfair monopolization.

Sec. 73. Publication. - The Board shall maintain its own publication which shall be known as the Plant Variety Gazette for all the publication requirements of this Act and for other purposes which the Board may require. Copies shall be distributed to all concerned especially to the Members of the Senate and House Committees on Agriculture: Provided, That the Board shall distribute for free, and in the major dialect understood by the locality, copies of the Plant Variety Gazette to small farmer groups and indigenous communities.

Sec. 73. The Registrar. - There is hereby established a National Plant Variety Protection Registrar and an Associate Registrar under the control and supervision of the Board. The Registrar and the Associate Registrar shall be appointed by the President of the Philippines upon the recommendation of the Board and shall have a term of six (6) years. However, the Registrar who shall be first appointed shall serve for a term of seven (7) years.

The Registrar shall be a citizen of the Philippines with good moral character, proven track record in the field of plant science, and/or extensive executive experience and capability.

Functions of the Registrar. The Registrar shall have the following functions:

a) Has original and exclusive jurisdiction to receive, process, examine all applications for Certificate of Plant Variety Protection in accordance with this Act, and in meritorious cases, issued the said certificates and sign them in the name of the Board;

b) Issue and maintain a systematic record of all Certificate of Plant Variety Protection and transactions related thereto;

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c) Implement the rules and regulations issued by the Board;

d) Institutionalize, maintain and continuously update a database of existing plant varieties collected from foreign and local databases;

e) Maintain a library of scientific and other works and periodicals, both foreign and local, to aid his examiners in the discharge of their duties;

f) Maintain samples of the propagating materials of the protected variety; and

g) Perform such other functions as may be prescribed by the Board.

TITLE XII

Miscellaneous and Final Provisions

Sec. 75. Relation with Other Laws. - The interpretation of the provisions of this Act shall not negate the effectivity and application of Republic Act No. 8371, otherwise known as the "Indigenous People's Rights Act"; Republic Act No. 9147, otherwise known as the "Wildlife Resources Conservation and Protection Act"; Presidential Decree No. 1151, otherwise known as the "Philippine Environmental Policy"; and Executive Order No. 430 and Administration Order No. 8, Series of 2002 of the Department of Agriculture or the rules and regulations for the importation and release to the environment of plant products derived from the use of biotechnology.

Sec. 76. Transitory Provisions. - The National Seed Industry Council, which was created by Republic Act No. 7308 or the National Seed Industry Development Act, shall perform the functions of the Board until the latter has been fully organized, but not later than three (3) years from the effectivity of this act. Within the same period, the Director of the Bureau of Plant Industry shall be the Acting Registrar and the Assistant Director of the same Bureau shall act as the Associate Registrar.

Sec. 77. Appropriations. - The Secretary of the Department of Agriculture shall immediately include in its program and issue such rules and regulations to implement the provisions of this Act, the funding of which shall be included in the annual General Appropriations Act.

Sec. 78. Separability Clause. - If, for any reason, any provision of this Act is declared invalid or unconstitutional, the other parts not affected thereby shall continue to be in full force and effect.

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Sec. 79. Repealing Clause. - All laws, decrees, executive orders, and rules and regulations, or parts thereof that are inconsistent with the provisions of this Act, are hereby repealed or modified accordingly.

Sec. 80. Effectivity. - This Act shall take effect thirty (30) days after its complete publication in a newspaper of general circulation.

Approved:

(Sgd) (Sgd)

JOSE DE VENECIA, JR.Speaker of the House of

Representatives

FRANKLIN M. DRILONPresident of the Senate

This Act which is a consolidation of Senate Bill No. 1865 and House Bill No. 4518 was finally passed by the Senate and the House of Representatives on May 30, 2002.

(Sgd) (Sgd)

ROBERTO P. NAZARENOSecretary General

House of Representatives

OSCAR G. YABESSecretary of the Senate

Approved: June 7, 2002

(Sgd)

GLORIA MACAPAGAL-ARROYO

President of the Philippines

GLADYS ANNE T. ZUBIRI..

Uhmmmm.. una kong nakita yung name niya sa DEAN’S LISTERS List.. siya yung second sa highest… ang kaso yung highest naman hindi nagchem.eng so ibig sabihin, si Gladys Anne Zubiri na yung may pinakamataas na GWA sa amin that time.. well, siya rin yung plging highest sa mga exams nmin kay Mam Cornelio.. galing kasi niyan sa Eng. Ana e.. sobra.. nakakaperfect siya sa mga exams.. tas silang dalawa ni Kuya Arjay S. Arnao yung huling-huling natanggal sa dean’s list..

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Maiba nmn ako.. hay.. ang lahat nakilala si ate gladys as tahimik and sobrang bait na bata.. yung tipong, ang mga kinukonsider niya as major na ngawa niyang kasalanan is sobrang minor lang para sa atin.. haha.. sa pagiging mabait, wala akong reklamo diyan.. pero yung sasabihing tahimik siya.. ako na nagsasabi sayo… isa yang kasinungalingan!!!! Hahahaha.. ang dami kaya niyang kwento, ang daming alam.. at mahilig bumanat ng mga jokes.. khit minsan mejo korni pinipilit niya pa rin.. (haha!!! frustrated joker kasi eh.. hilig niya lng din talgang mapasaya yung mga taong mahalaga pra sa kanya kaya ganun..) at kapag malakas ang trip niyan kung anu-ano yung ginagawa niya…tulad ng mga bagay na hindi ginagawa ng mga normal na tao.. haha.. (halimbawa: seryoso kayong naglalakad sa kalye bigla-bigla na lng yan maga-act as traffic enforcer.. and the likes.. yung mga tipong hindi mo ma-imagine na gagawin niya) tsaka tulad ni ate Kathleen May E. Dayao na double chin.. marami din siyang mga bagay na nasasabi kapag bangag siya… (halimbawa: ang tawag niya sa taong nawalan ng memories ay may ANESTHESIA..) kaya naman gaya nga ng sbi ni kuya Kim Bryan L. Duenas “bakit ka tahimik? Hindi ako sanay e”.. ayun, kaming mga usual na kasama niya ay hindi sanay na tahimik siya.. at parang there’s something wrong kapag ganun.. tsaka siguro gusto niya rin maging kasing galing ni kuya Bernard F. Galangue sa pagdu-drawing kasi kapag kakaiba na naman yung trip niya, habang may hawak na ballpen at papel sasabihin niyan sayo wag kng gagalaw, kala mo naman iisketch ka.. yun pala isusulat lng yung pangalan mo.. lakas mag-trip…hahaha..

Ayun.. sobrang humble ng batang yan e.. as in.. sobrang dina-down niya yung sarili niya.. and as a friend, wala akong masasabi.. taas kamay ako diyan.. at kahit sino pang friend ang tanungin mo.. isa lang ang sasabihin sayo.. the BEST siyang friend.. lagi mo yan malalapitan kapag kailangan mo ng tulong at kahit hindi ka pa humihingi ng tulong siya na yung mag-ooffer ng help.. haha.. astig yan sa lhat ng bagay.. kaya IDOL ko siya.. hindi lng ako kundi kaming lhat…

By Jennifer Grace Jamero

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SINO SI KAKAI?

Sa totoo lang, hindi alam ng karamihan na mahilig kami ng pamilya ko gumawa ng funny faces. Mukha lang akong seryoso pero minsan kalog din ako. Minsan lang… Hehehe… ^_^

Ayan, hindi kilala ng karamihan yang mga taong yan except kay Chichi (leftmost). Chichi is my younger sister, andami sa mga classmates ko ang nagkagusto at nagkakagusto riyan since highschool pa. Tapos yung nasa gitna, sabi nila yan daw ang tunay kong ina. Hehehe… Hindi. Ninang ko yan; Si Ninang Tina. Kapatid ng Daddy ko. Tapos yung nasa rightmost ay yung ka-live in ng Daddy ko. Bale second mom namin; Si Tita Gigi. Sa katunayan, kahit sino kina Ninang Tina at Tita Gi pwedeng pumasa bilang tunay kong ina sa sobrang close ng resemblance namin.

Wala lang. Sample picture lang dun sa trabaho ko nung summer of 2010. Na-enjoy ko kasi talaga yung dive nun sa Batangas kahit

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three days lang yun. Hehehe… ^_^ Super over-rated na pero gusto ko yung magiging trabaho ko in the future may kinalaman sa environment conservation. Kahit di na environmental engineer pero ayoko maging perwisyo kay mother nature as much as possible. (Eew… Corny…)