Download - The Alliance in the - DSM Feedback
- Introduction to the DSM-Novozymes Feed Enzymes Alliance
• Who are Novozymes and DSM?
• What is the Alliance? Why did we decide to create it?
• How do we work together to bring innovation to the industry?
• What is our position in the market?
- Value we generate for the Animal Nutrition industry
• What are the key challenges the animal industry is facing?
• How are we addressing these challenges?
• How do we generate value for our customers?
Welcome!
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DSM at a glance
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Sales by Destination
Europe
33%
Americas
41%Asia
21%
ROW
5%
Sales by End-Market
Personal
care
3%
Animal Nutrition& Health
48%HumanNutrition& Health
38%
Food specialties
11%
Sales*
>€10 billion
Global* presence
~25,000 employees worldwide
Listed at Euronext NYSE
Share price ~ quintupled in 25 years
Sustainability leader
Top rankingDow Jones
Sustainability Index
Sales distribution in segmentsAround the world• A pioneer from the earliest days of feed additives,
we are one of the world’s leading suppliers of
vitamins, carotenoids, eubiotics and feed
enzymes to the global feed industry.
• We are successful in this sector because we
provide our customers, such as feedmills,
integrators and farmers with products and
solutions that help them succeed in a dynamic
and ever-changing global market, enhancing both
profits and environmental sustainability.
vitamins
carotenoids
eubiotics
feed enzymesOther
solutions
DSM’s Animal Nutrition & Health Business- A global force
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Page 9
Enzymes research & patents
Form development & production
Application & stability testing
Marketing, sales, distribution & premix
• + 1,000 R&D employees• Largest microorganism and
enzyme collection• Enzymes screening • Protein engineering• Formulation development• …
No. 1 in Bio-Innovation
• In vivo trials • In-feed analytics• Scientific Network• Global marketing, sales
and distribution • Laboratory service • …
No. 1 in Animal Nutrition
Two leaders joined forces 20 years ago - Full commitment to success as “One Company”
Bringing significant Innovations to Industry- To get more out of feed ingredients and reduce costs
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2004 201420062000
201520112008
Challenges in the animal nutrition industry
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1.
World population
expected to reach 9+
billion by 2050
2.
Changing diets are
expected to double the
demand for protein over
the next 40 years
4.
Consumers desire
healthy food at an
affordable price
3.
Arable land per capita
expected to decrease by
35% from 2000 to 2050
5.
Consumers want healthy
animals grown without
antibiotics
6.
How to make our
industry more
sustainable?
In 2016 the Alliance and our customers reduced the Carbon Foot Print by 3.1 Million Tons of CO2 Eq.
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Number of incandescent lamps switched to LED 109,512,609
Number tree seedlings grown for 10 years 80,123,632
Number of passenger cars driven for one year 1,288,990
Equivalent to:
We are focused on Sustainable Animal Nutrition
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• Care for safe and controlled
animal production
• Continuous drive to raise animal
health
• Bring innovations to increase
animal nutrition sustainability
• Combat antibiotic resistance
• Improve gut health
• Reduce use of scarce resources
• Reduce emissions from livestock
• Reduce food waste
• Advocate science-based
knowledge about animal protein
• Leverage joint expertise
• Improve end product quality and
food safety
We work in customer centric approach to deliver local solutions with our global products
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+
Key Enablers
Global
Products
Local
Solutions
Actives Forms Premix
DSM Actives DSM Forms
Novozymes
Enzymes
Novozymes
Forms
SolutionsAccess &
Insights
• The enzyme development process
• Phytases
- Lars Kobberøe Skov
• Proteases
- Paulina Tamez-Hidalgo
• Starch degrading enzymes
- Morten Tovborg
• Fiber degrading enzymes
- Ninfa Rangel Pedersen
• Sustainability
- Per Henning Nielsen
Introduction to
The world of enzymes
Enzyme development process
For feed enzymes we have to studythe physiology of the animals
What are the conditions in the different parts of the
digestive tract? Where can the enzymes work?
What is the substrate? …. And under what
conditions should the
enzyme work?
What kind of stability issues can
be envisaged ?
How can I make it into a product?
How…. ?
Enzyme development process
1) Create diversity to have many options
to choose from
2) Test candidates to select best enzyme
2) Develop production process
Create diversity to have many options to choose from
Protein engineering
Genes in environmental samples
Sequence information
Natural (wild type) microorganisms
SWISSPROT:C7Z624 DNSGYCLKDRKQ-KCECFAGFTGSKCDKYTCVDKCSGHGKCVGPNECKCNKGWGGLHCSF
SWISSPROT:B6HRY4 QENGFVDGDGSL---ECFTGFTGTDCTQFTCPNSCSGNGKCVGPNECKCEDSWAGPECSW
SWISSPROT:C7Z624 LLIEPTYETESRLGDGDDPAIWISPESPEKSRVVTTMKSGKEAGLGVFDLAGNLLQSFPA
SWISSPROT:B6HRY4 VGVEPKYETDANGGDGDDPAIWISPVSADQSTIITTTKSELGAGFAVFDLAGKLLQTVSA
Phytase - definitions, uses and benefits
Phytic acid
• Phosphorous storage in plants
• Indigestible by mono-gastrics
Phytase
• Catalyzes the hydrolysis of phytic acid
• Used in animal feed to release phosphate
• Reduces need for addition of inorganic phosphate
Benefits
• Reduced feed costs
• Reduced levels of phosphorus released to the environment
Phytate
• ‘Salt’ of phytic acid
• Negatively charged phosphate groups
• Complexing agent
Inositol hexakisphosphate (IP6)
Phytatedegradation
H2OPi
H2OPi
H2OPi
H2OPi
H2OPi
IP6
IP5
IP4
IP3
IP2
IP
Phytases are
a special group
of phosphatases!
Selection processIn vitro test : Analysis of phytate and degradation products
0.0 5.0 10.0 15.0 20.0 25.0 32.0
-0.100
-0.000
0.100
0.200
0.300
0.400
0,500
0.600
0.700
Ab
so
rbance, 290 n
m
Time (minutes)
1
2
6
5
3
4
Ion
-ex
ch
an
ge
co
lum
n
IPs
0
1
2
3
4
5
6
7
8
0 5 10 15
pH
Incubation time (minutes)Phytate is
extracted from
the sample
using HClElution as follows:
IP1 → IP2 → IP3 → IP4 → IP5 → IP6
Fe(NO3)3
Feed +
enzyme
Stomach
+ pepsinCrop
Detection of IP-Fe complexes
RONOZYME® HiPhos
• produced by an Aspergillus oryzae strain
expressing a synthetic gene coding for the
phytase from Citrobacter braakii. (Lichtenberg et al., 2011)
• increases the amount of plant phosphorus
available to the animal
• substantial savings on feed costs
• Increases weight gain and feed conversion
because more phytate, an antinutritional
factor, is broken down
• reduces environmental impact
Proteases – definitions, uses and benefits
Protein
• Chains of amino acids linked
by peptide bonds
• Important for various metabolic
processes, including building
muscle tissue in livestock
• The most costly part of animal
feed rations after energy
Protease
• Catalyzes the hydrolysis of
peptide bonds thereby
degrading proteins
• Used in animal feed to
increase digestibility of protein
Benefits
• Reduced feed costs
• Reduced levels of nitrogen
released to the environment
FunctionalStructural
Globular ProteinsAlbumins, Globulins, Prolamins,
Glutelins, Glycinins and Enzymes
Water soluble
Fibrous ProteinsKeratin, Collagen, Elastin,
Actin, Tubulins
Water insoluble
Proteins can be divided intotwo main categories
• Approx. 2% of proteins are proteases
• Proteases have different functions, including processing of proteins and protein turnover
Proteases degrade proteinby specific cleavage patterns
Endo-peptidases
NH2+ COOH
NH2+ COOH
Endo-peptidasesEndo-peptidases
NH2+ COOH
NH2+ COOH Exo-peptidasesNH2
+ COOH
NH2+ COOHNH2
+ COOH
NH2+ COOH Carboxy-peptidasesAmino-peptidases
Diversity of digestive proteases
Cysteine
Metallo
Serine
Threonine Aspartic
Pepsin
Pancreatic proteases
• Trypsin
• Chymotrypsin
• Elastase
• Carboxypeptidases A & B
RONOZYME® ProAct
RONOZYME® ProAct is produced by Bacillus licheniformis
expressing a gene from Nocardiopsis prasina encoding a
serine protease of subfamily S1
Glitsoe et al., (2012)
RONOZYME® ProAct is a bacterial serine protease, which is safe for use in the animal feed industry
• The enzyme product is manufactured by fermentation of microorganisms that are not present in the final product
• The production organisms are improved by means of modern biotechnology
RONOZYME® ProAct enables reduced protein levels, minimizing the nitrogen excretion
Feed formulation
N
N
N input
N input
N output
N output
Consumption of protein
Protein in manure
Reduced N consumption
Reduced manure N content
No change in N content of the chicken
Ronozyme® ProAct
RONOZYME® ProAct is a unique protease that brings both economical and environmental value
Protein
• Chains of amino
acids linked by
peptide bonds
• Important for various
metabolic processes,
including building
muscle tissue in
livestock
• The most costly part
of animal feed rations
after energy
Protease
• Catalyzes the
hydrolysis of peptide
bonds thereby
degrading proteins
• Used in animal feed to
increase digestibility of
protein
RONOZYME® ProAct
• Unique & pure microbial protease product that is pelleting
and gastric stable
• Reduces feed formulation costs
• Reduces levels of nitrogen released to the environment
• Works on broad range of feed ingredients
• Compatible with other feed enzymes
• Potential tool to help combat marginally high ANF levels in
SBM
Amylases – definitions, uses and benefits
Starch
• Insoluble crystalline polymer of
amylopectin and amylose, made
of glucose
• Synthesized by most plants for
storage of energy and
carbohydrate building blocks
• Starch is less crystalline than
cellulose, and the bonds
between glucoses are
hydrolyzed more easily
Amylases
• Catalyzes the hydrolysis of
alpha-1,4 glycoside bonds in
starch to form small soluble
dextrins
• Found in many organisms from
bacteria and fungus to higher
animals
• Used in starch and bioethanol
industries and in animal feed to
improve starch degradation
Benefits
• Improves rumen starch
hydrolysis
• Improves fiber degradation by
cross-feeding fiber degrading
bacteria
• Increases energy release,
resulting in improved milk
efficiency
Starch degradation
Amylopectin
Beta-amylase
Glucoamylase
Isoamylase
= Hydrolysis of -1,4-linkages
Amylose
Maltose
Glucose
Alpha-amylase
= Maltose
Amylase selection
Traits of an amylase for
ruminants
• Synergistic starch activity
• Stability towards proteolytic
enzymes
• Stability and activity at pH-ranges
from rumen to duodenum
• Pelleting stability
Synergistic starch activity with endogenous amylase
In vitro digestion of corn starch containing diet (pelleted)
Pancreatic amylaseRumiStarTM +
pancreatic amylase
RONOZYME® RumiStar selection
0
20
40
60
80
100
120
pH3: 0min
pH3: 45min
pH3: 120min
pH4: 0min
pH4: 45min
pH4: 120min
Res
idu
al a
cti
vit
y (
%)
0
20
40
60
80
100
pepsin (pH3) pepsin (pH4) pancreatin(pH7)
ProAct (pH7)
Res
idu
al a
cti
vit
y (
%) 95,7
89,8
93,7
85,7
0
10
20
30
40
50
60
70
80
90
100
After mixing(mash)
Afterpelleting
Pellets 1month
Pellets 3months
Recovery
(%
)
Proteolytic stability
pH stability
Pelleting stability
Traits of an amylase
for ruminants
• Synergistic starch activity
• Stability towards proteolytic
enzymes
• Stability and activity at pH-
ranges from rumen to
duodenum
• Pelleting stability
RONOZYME® RumiStar- mode of action
RONOZYME® RumiStarTM
is a pure α-amylase selected for ruminal conditions.
It catalyzes the hydrolysis of starch to oligosaccharides in the rumen.
It optimizes ruminal starch digestion, increasing VFA.
Fibrolytic bacteria profit from the enzymatic starch breakdown (cross-feeding), increasing the ruminal fiber degradation.
Improved feed degradation provides additional energy for higher performance.
RONOZYME® RumiStar 600 CT
• Bacterial thermostable alpha-amylase
produced by Bacillus licheniformis
• Selected for ruminal conditions
• Optimizes starch utilization
• Improves milk efficiency
€ milk - € feed
Fiber-degrading enzymes –definitions, uses and benefits
Fiber
• Non-starch polysaccharides
(NSPs)
• NSPs are resistant to
digestion in mono-gastrics and
hence anti-nutritional
• Main fiber components in
cereals and oilseeds differ
Fiber-degrading enzymes
• Degrade soluble NSPs
thereby reducing viscosity
• Degrade insoluble NSPs
thereby releasing nutrients
trapped within cell walls
• Produce oligomers
Benefits
• Increase nutrient absorption
and feed intake by decreasing
digesta viscosity
• Increase bioavailability of
nutrients such as starch and
protein
• Potential prebiotic effect of
oligomers
Milling and digestion does not release all starch and protein in cereals
Milled corn stained with iodineLight microscopy
Jejenum samples from chickens fed corn stainedwith iodine Light microscopy
The process of oil removal does not destroy cell walls
• Intact cell walls
in protein meals
• Protein is
trapped within
the cells
Sunflower
Sunflower meal Rapeseed meal Soybean meal
SoybeanRapeseed
Main NSPs in cereals
.
Main NSPs in oilseeds
RONOZYME® WX and RONOZYME® Multigrain RONOZYME® VP and RONOZYME® Multigrain
• Arabinans
• Homogalacturonan (pectin)
• Rhamnogalacturonans
• Xylogalacturonan
• ß-1,4-glucan
• Xyloglucans
• Mannans (Galactomannans)
• Galactans (Arabinogalactans)
• ß-1,4-glucan
• ß-1,3-1,4-glucans
• Xylans (Arabinoxylans)
Model of plant cell wall (mono-cotyledon)
Mainly consists of:
Carpita, N.C. and Gibeaut, D. M (1993). The Plant Journal 3(1):1-30.
A mixture of polysaccharides interlocked in a complex structure
1. Arabinoxylan
2. Cellulose
3. Beta-glucan
A mixture of polysaccharides interlocked in a complex structure
Model of plant cell wall (di-cotyledon)
Cellulosefibrils
Hemicellulose: Xyloglucan
Galactomannan
Pecticpolymer matrix
Carpita, N.C. and Gibeaut, D. M (1993). The Plant Journal 3(1):1-30.
Three independent networks:
1. Cellulose, hemicellulosenetwork
2. Pectin network(≈ 60% of the polysaccharides)
3. Structural proteins
RONOZYME® WX
A xylanase from
Thermomyces
lanuginosus produced by
Aspergillus oryzae
RONOZYME® Multigrain
A non-GM product
containing xylanase and
β-glucanase produced by
Trichoderma reesei
RONOZYME® VP
A non-GM product
containing β-glucanase,
hemicellulase and
pectinase produced by
Aspergillus aculeatus
Our fiber degrading enzymes: sources
Wheat slice: Image immediately after additon of RONOZYME® WX
Wheat: Image 30 minutes after addtion of RONOZYME® WX
Wheat: Image 60 minutes after addition of RONOZYME® WX
Wheat: Image 120 minutes after additon of RONOZYME® WX
Wheat: Image 180 minutes after addition of RONOZYME® WX
Degradation of arabinoxylan cages can be visualised by microscopy - fx by autofluorescence of ferulic acids
Wheat slice, control, 400XBarley slice, control, 400X Rye slice, control, 400X
RONOZYME® WX
Cereal cell wall degradation visualizedby SEM
Slices of wheat, rye and barley incubated +/- enzyme (1 h, 40oC).=> Cell walls visible in control samples; enzyme treatment results in empty space where the cell walls used to be => bioavailability of starch and protein
Most important protein sources in animal feed areSBM and canola meal
Rapeseed/canola meal(RSM)
Soybean/soybean meal(SBM)
1
10X SBM, 3 h with VP (100X dilution),
390CControl , 10X magnification SBM, 10X SBM, 1 h with VP (100X dilution),
390C
Soybean
RONOZYME® WX
RONOZYME® Multigrain
RONOZYME® VP
• Better performance and lower feed costs
• Improves digestibility of a wide variety of feeds
by reducing intestinal viscosity and releasing
nutrients, as well as offering a potential
prebiotic effect of oligomers
Global challengesPhosphate depletion -
risk of reduced yields in
agriculture
Climate change and risk of rising
sea levels
Nutrient enrichment –
risk for fisheries
UN, 2015
10 billionpeople in
2050
pause
Greenhouse gas emissions caused by humans
Source:
Food and Agriculture Organization of the United Nations (FAO) 2013
FAO recommendations
• Reducing land-use change from feed crop cultivation
• Improving the efficiency of crop production
• Improving the efficiency of energy generation and supply
• Reducing the use of uncovered liquid manure management systems
• Improving feed conversion of the individual animal
• Providing balanced animal nutrition
Source:
Food and Agriculture Organization of the United Nations (FAO) 2013
Enzymes improve feed conversion
• RONOZYME® HiPhosimproves phytate digestion
• RONOZYME® WXimproves xylan degradation
• RONOZYME® ProActimproves protein digestion
Good for the planet and the farmer
Impact of RONOZYME® HiPhos - phytase
Natural phosphorus in
the feed is released
5%
Algae bloom is
reduced
Save inorganic
phos-phorus
Phos-phorus in manure is reduced
Phos-phorus in lakes and rivers is reduced
Source: Nielsen and Wenzel (2006)
Impact of NSP enzymes; for exampleRONOZYME® WX
Less
manure with
less N
Less
N2O and
CH4
Less NO3
and NH3
Reduce climate change
Reduce algae bloom
Source: Nielsen et al. (2008)
Saves feed and replaces some soy, fat
and wheat with
barley
Impact of RONOZYME® ProAct - protease
Saves Soy
Bean Meal
Less
nitrogen
manure
Less
N2O
Less
NO3- and
NH3
Reduce climate change
Reduce algae bloom
Source: Oxenbøll et al. (2011)
Perspectives
Use of xylanase for pigs can
reduce greenhouse gas
emissions by 3-8%
~ 4 million MT CO2 avoided
annually in Europe
Use of protease for poultry can
reduce ammonia emissions
by 3%
~ 12,000 MT NH3 avoided
annually in Europe
Use of phytase can almost
eliminate supplementation of
inorganic phosphorus to
pigs’ feed.
Source: Nielsen et al. (2008)Source: Oxenbøll et al (2011); DSM
sustainability calculator
Source: Nielsen et al. (2017). In prep.
~ 30,000 MT phosphate rock
saved annually in Denmark