The Alliance in the

School Yard

José Otávio Berti-Sorbara (DSM)

Max Josef Jäger (NZ)

- 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!

3

Novozymes’ enzymes improve production efficiency, save energy and raw materials

4

Novozymes’ products impact your everyday life

5

Novozymes is the leader in industrial enzymes

6

14,142

DSM at a glance

7

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

8

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

10

2004 201420062000

201520112008

Challenges in the animal nutrition industry

11

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.

12

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

13

• 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

14

+

Key Enablers

Global

Products

Local

Solutions

Actives Forms Premix

DSM Actives DSM Forms

Novozymes

Enzymes

Novozymes

Forms

SolutionsAccess &

Insights

Introduction to

The world of enzymes

• 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

The enzyme

development process

Lars Kobberøe Skov

Protein Biochemistry & Stability

Novozymes R&D

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

Phytases

Lars Kobberøe Skov

Protein Biochemistry & StabilityNovozymes R&D

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!

The microbialworld of HAP phytases

P. lycii

C. braakii

Figure from Ariza et al., (2013)

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

Paulina Tamez-Hidalgo

Feed Applications

Novozymes R&D

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

Proteins fold up in complex structures

50% of our cell molecules are protein

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 protein byhydrolysis of peptide bonds

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

Starch degrading

enzymes

Morten Tovborg

Feed ApplicationsNovozymes R&D

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

No 1

Gallant et al 1997

Non-structural carbohydrate

The structure of starch

Starch degradation

Amylopectin

Beta-amylase

Glucoamylase

Isoamylase

= Hydrolysis of -1,4-linkages

Amylose

Maltose

Glucose

Alpha-amylase

= Maltose

Amylase selection

Mammalianamylases

RONOZYME®

RumiStarTM

GH13 with subfamilies

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

enzymesNinfa Rangel Pedersen,

Feed ApplicationsNovozymes R&D

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

Dietary fiber

Cereal

Starch

Cell walls

(fiber, non starch

polysaccharides)

Fiber

Protein

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

RONOZYME® WX

(GH11 xylanase)

Degradation of fiber by RONOZYME® WX

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

Sustainability

Per Henning Nielsen

Corporate SustainabilityNovozymes

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

Thank you