12 industrial fermentation 2008 ho
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Industrial fermentation Production with microbes Industrial microbiology
The use of microbes in the production
Start from an ancient time
Large scale production
Early 20th century beverage industry, vinegar,
baking yeast, citric and lactic acid
Late 20th century development of
biotechnology from the antibiotic industrial
Production with microbes
Fermentation industry
Microbes to produce useful product
Usually a naturally produce substances for
growth, and maintenance.
Metabolites
Primary metabolite produce during growth
Secondary metabolite produce after growth
stage
Substrate
Product
Cell
Time
Substrate
Product
Cell
Time
Primary metabolite
Secondary metabolite
Trophophase Idiophase
ProductsI. Foods and foods related- Fermented meat, Cheeses and milk products,mushrooms, bakers yeast, coffee, pickles, singlecell-proteins, vinegar, amino acids, vitamins,alcoholic beverages
ProductsII. Organic acids- Citric acids, Itaconic acid
III. Enzymes and microbial transformation
- Commercial enzyme, sterol conversion
IV. Engineered microbes
- Insulin, human growth factor
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Production with microbes
Steps in Industrial fermentation
Isolation of microbes that produce your
interested product
Screen for the best producing strain: naturally or
mutation.
Optimize production condition (growth
basically) in lab
Scale up from lab scale (up to 10 L) to industrail
scale (>10,000 L) (raw material?)
Food and food related products
Whole cells products
Including mushrooms and Single-cell protein
(SCP) Mushrooms fruiting body of mold!
Highly consumed every year high nutrition.
Grow on decaying organic matter in soil or wood.
Stalk and cap = compact mycelium
Gills = site of reproductive spores
E.g. Agaricus bisporus(botton mushroom),
Lentinus edulus(Shitake mushroom)
Agaricus bisporusFood and food related products
Single-cell protein yeast, and cyanobacteria
Alternative food sources mostly used now as
supplementary diet e.g. yeast is a source for B
Vitamins.
Organisms: yeast, Spirulina
Growing biomass and freeze dry Raw material from other industrial, farming
Food and food related products
Bakers yeast Same process as in single cell protein
Food and food related products Cheese production
Can produce from any milk.
4 phases: coagulation, separation, shaping
and ripening
Coagulation: rennin enzyme or acids precipitatecasein
From calvess stomachs, engineered bacteria
Acid production by lactic acid bacteria
Curd entrap bacteria, fat globules and othermaterial.
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Cheese production
Separation & Shaping
Additives (salt and herbs) may be added after seperation
or shaping: effect property of cheese product
Ripening add flavor and look
Natural or inoculation of microbes e.g. Penicillium
roquefortii (spores are usually added into blue
cheese), and Penicillium camembertiis added to
the surface of a cheese known as Camenbert
Blue cheese
Brie
Gorgonzola
Swiss
Food and food related products
Yogurt and other fermented milk products
Yogurt, kefir, buttermilk, sourcream
Yogurt usually use mixture of 2 cultures:
Streptococcus thermophilus: ini. acid production
Lactobacillus bulgaricus
0.5 - 1.0% lactic acid
Fermented at 28-32 C
Taste due to acetaldehyde, diacetyl, and acetoin
Product need to contains live cultures
A mixed lactic acid and
alcoholic fermentation.
Streptococcus lactis,
Lactobacillus
bulgaricus, yeasts
Kefir
This milk product is used for its
medicinal therapeutic value.
Lactobacillus
acidophilusAcidophilu
s milk
Made with skimmed or partly
skimmed pasteurized milk.
Streptococcus sp.
Leuconostoc sp.Cultured
buttermilk
Cream is inoculated and leave
to develop acidity
Streptococcus sp.
Leuconostoc sp.Sour cream
DescriptionMicroorganismsFermented
Product
Some Foods Produced from Fermented Milk.
Industrial production
of yogurtKefir Buttermilk
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Food and food related products
Alcoholic beverages
Accidental discovered from ancient human
Saccharomyces cerevisiae Raw materials are regional.variety of products
Wine grapes, fruits
Beer cereal grain barley, sorgum, wheat
Other rice wine (rice), teguila (agave cactus),
vodka (corn), mead (honey).
Food and food related products
Fermented meat
Use to preserve food E.g. salami, summer
sausage, cured ham
Thailand naam (fermented pork), fermented
sausage, fermented fish, fish sauceetc
Main organism Pediococcus serevisiae,
Lactobacillus plantarum, other lactic acid bacteria
Production of lactic acid to change pH of food
Production of probiotics in some strains
Food and food related products
Fermented fruits and vegetables
Found around the world
With bacteria, yeast and mold
Depends mainly on native microbes
associate with the plants.
Most included mixed culture
fermentation
Conditioning by human to favor growth
of beneficial microbes - salting
http://www.coffeeresearch.org/Pictures/fermentationtanks.jpg
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Rhizopus oligosporus, Rhizopus oryzae,
Lactobacillus delbrueckii, Saccharomyces
rouxii
Soy sauce
Leuconostoc mesenteroides,
Saccharomyces marscianus,
Flavobacterium spp., Fusarium spp.
Coffee
Lactobacillus mesenteroids, Lactobacillus
plantarum
Sauerkraut
Pediococcus cerevisiae, Lactobacillus
plantarum
Pickles
Leuconostoc mesenteroides, Candida sp.Cocoa products
OrganismsProducts Food and food related products Flavoring agents: amino acids and nucleotides
Monosodium glutamate, inosinic acid flavor
enhancers
Originally use seaweed, fish
Produce by enzymatic hydrolysis of yeast RNA
(Candida utilis)
Now direct fermentation = monosodium glutamate
and 5-IMP (inosine monophosphate)
Brevibacterium ammoniagenesproduce inosine and
then chemically phosphorylated
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Non-food products
Enzyme production
Produce in large scale around the world
Major producer Novozyme (44%), GenencorIntl, USA (18%)
Cheese productionMicrococcusLipases
Fruit juice clarificationAspergillusPectinase
Cheese manufactureAlcaligenes,
Aspergillus,Candida
Rennin
Starch liquefaction, brewing,
baking, feed, detergents
BacillusBacterial amylases
Fructose syrup productionAspergillusGlucoamylase
Antitumor agentEscherichia,
Serratia
Asparaginase
SweetenersBacillus, StreptomycesGlucose isomerase
Semisynthetic penicillinsEscherichiaPenicillin acylase
DetergentsBacillus, StreptomycesBacterial proteases
UseOrganismsEnzyme Non-food product
1. Vitamins 2. Antibiotics
3. Alkaloids 4. Steroids
5. Non-Steroid Hormones/cell regulators (cytokines):
1. Epidermal growth factor 2. Proinsulin
3. Insulin 4. Human growth hormone
5. Somatostatin 6. Interferons
7. Platelet-derived growth factor
8. Fibroblast growth factor 9. Tumor Necrosis Factor
10. Other cytokines are coming on line all the time
Non-food product6. Blood coagulating factor XIII
7. The restriction enzymes
8. Other enzymes (e.g. ligase, DNA polymerases etc.)
9. Solvents
How do the microbes produce these products? Genetically modified microorganisms
Clone genes that encode protein of interest into
vectors and express this genes in microbes used
in production.
Able to produce high amount of compounds than
in original organisms with ease of manipulation.
Microbes able to utilize waste from other process
as a raw material for fermentation.
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Antibiotics
Penicillin by Penicillium chrysogenum
Now most of the antibiotics were discover from soil
microorganisms especially in Streptomyces spp. Engineered genes into production strains.
Modification of existing antibiotics.
http://www.sanger.ac.uk
http://scienceblogs.com
Streptomycin from Streptomyces griseus
http://www.agen.ufl.edu/~chyn/age4660/lect/lect_21/f15_11.JPG
Alkaloids
Nitrogen containing organic compounds that
derived from plants
Have medicinal purposes:
Atropine dilates pupils of the eyes (some muscle
disease)
Morphine and codeine relief of pain
Cocaine local anesthetic
Quinine, caffeine, nicotine, strychnine, serotonin , and
LSD
Atropineserotonin
Steroids Types of products
include cortisone
and reproductive
hormones
Human proteins
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Biotech Drugs
Mainly recombinants proteins and monoclonalantibodies.
Highly active fields of biotech
Lots of new products each year
For list of product approve in US www.bio.org
Timeline in drug development
top related