The Role That Enzymes Can Play In Terms Of Increasing The Efficiency By Which Animals Convert Feed Into Protein

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The Role That Enzymes Can Play In Terms Of Increasing The Efficiency By Which Animals Convert Feed Into Protein Dr. Charlotte Horsmans Poulsen, Senior Staff Scientist, DuPont Industrial Biosciences 1

What I Will Discuss Today Food Security Index How enzymes» Can help the industry to produce more protein» Enable the use of new, alternative raw materials Future alternative feedstocks (and other) technologies» Which might change in the industry Summary

The DuPont Global Food Security Goals By the end of 2020, DuPont will help the world meet the challenge of achieving global food security Innovating to Feed the World We will commit $10 billion to R&D and 4,000 new products will be introduced. Engaging and Educating Youth We will facilitate two million engagements of young people around the world in educational opportunities. Improving Rural Communities We will work to improve the livelihoods of at least three million farmers and their rural communities through targeted collaborations and investments. 3 3

The Global Food Security Index DuPont commissioned the Economist Intelligence Unit in 2012 to develop the Global Food Security Index Ranks 109 countries according to their relative levels of food security using 29 indicators divided into three categories: Affordability, Availability, Quality and Safety. Provides a rigorous, structured framework for understanding the drivers of food security. Visit foodsecurityindex.eiu.com

Protein Needs Are Increasing FAO* expects world demand for (animal-derived) protein to double by 2050 Increasing population (9 billion by 2050) Emerging economies Increasing urbanization Recognition of protein s role in a healthy diet Increased need for protein in the elderly population *FAO: Food and Agriculture Organization of the United Nations 5

6 Improvement in Slaughter Weight Versus Feed Conversion Ratios* 1.68 Kg 1.82 Kg 1.98 Kg 2.19 Kg 2.77 Kg 1960 1970 1980 1990 2011 1960 2.5 Kg 2.1 Kg 2.04 Kg 1.92 Kg 1.72 Kg *Amount of feed required for 1Kg weight gain Based on Rauw et al., 1998 and Ross info

Meat (kg/flock) Still Genetic Potential to be Captured Sub-optimal production Sub-optimal diet digestibility Feed ingredient variation Sub-clinical disease Clinical disease Environmental stress Production frontier Commercial performance Value Potential Reduced performance + mortality O Feed, labor, other inputs ($)

Cheaper Protein Sources Add Dietary Variability

Enzymes Work by Targeting Specific Substrates Substrate Effect of substrate Enzyme Soluble viscous NSPs (e.g arabinoxylans) Insoluble, non-viscous NSPs Starch Protein viscosity and digesta retention time nutrient absorption proliferation of intestinal microflora accessibility of nutrients by physical entrapment Metabolisable energy substrate for gut microflora Metabolisable energy and AA substrate for gut microflora (neg) Xylanase Xylanase Amylase Protease Lipid emulsification, digestibility of lipids Lipase Raffinose and stachyose Undigestible by animal enzymes α-galactosidase Beta glucan viscosity and digesta retention time β-glucanase Phytate Binds minerals, protein and starch Phytase

Exogenous Enzyme Solutions Diets To Reduce Variability What Positive Impact Does the Animal Get? Feed intake Digestion Fermentation Faeces Endogenous inputs a.a., NE, other Absorption SCFA a.a., NE Production Reduce endogenous inputs (a.a., NE): Degrade anti-nutrients Reduce physical interactions between digesta and gut wall Modulate gut microflora Change organ size Improve digestion of starch, fat, protein, minerals: Addition to endogenous enzymes New enzyme activities Remove physical entrapment Disrupt substrate interactions Modulate fermentation (SCFA): Production of pre-biotics Increase production of short chain fatty acids

Enzymes Reduce the Amount of Undigested Substrate Reaching the Lower Gut Ileal undigested starch (g/kg feed) Ileal undigested protein (g/kg feed) 14.3-43% 6.7-18% 6.0 5.5 8.5 8.1 Control XA XAP Control XA XAP Control XA XAP Control XA XAP Ileal undigested fat (g/kg feed) 4.6-20% 3.9 3.7 Control XA XAP Control XA XAP X: Xylanase; A: Amylase; P: Protease Adapted from Romero et al., 2012

Even Simple Diets Can be Variable in Quality The impact of xylanase, amalyse and protease addition to 56 different corn samples included in broiler diets reduced the variation in performance measured as FCR. (Danisco Animal Nutrition, 2011)

Wheat Variety Can Influence Pig Growth And Feed Intake 31% variation in daily gain between the best and next-to-worst Australian wheat samples FCR 1.18 1.15 1.14 1.10 1.11 1.13 1.16 1.15 1.09 1.14 502 514 476 486 432 433 438 445 438 447 419 432 388 396 399 394 376 341 233 271 1 2 3 4 5 6 Wheat variety 7 8 9 10 Growth: P<0.001 Feed intake: P <0.001 Feed:gain: NS Daily gain (g) Feed intake (g/day) 65% wheat in diets Weight 7-16 kg Source : Cadogan et al (1999)

Xylanase Reduces Variation in Performance Between Different Varieties of Wheat Daily gain (g) FCR: 1.38 1.27 1.14 1.19 1.17 1.19 Feed intake (g/day) Improvement with xylanase : 103% 5% 4% 75% -4% 9% 230 425 460 466 445 479 318 540 525 556 521 570 Wheat variety 1 5 10 1 5 10 1 5 10 1 5 10 + xylanase + xylanase Xylanase effects: Daily gain P<0.001 Feed intake P<0.001 FCR NS Wheat x Xylanase P <0.001 Source : Choct et al (1999)

Inorganic Phosphorus: A Non-Substitutable and Finite Resource http://nicholsoncartoons.com.au/phosphate-mines-in-vogue-226.html 15

Economically viable reserves (Gt) Inorganic Phosphorus: A Non-Substitutable and Finite Resource There are no substitutes for phosphorus in agriculture As world phosphate demand grow, both for animal feed and fertilisers, increasing price pressures will continue Global Inorganic Phosphate Reserves 6 Monthly rock phosphate prices ($/MT) 250 5 200 4 3 2 1 150 100 50 0 Morocco and Western Sahara China Jordan South Africa United States Other Countries 0 2009 2010 2011 2012 (U.S. Geological survey, 2010) (World Bank, 2012)

Phytate - Also Known As IP6 - Is The Natural Store Of Phosphorus In Plants But The Content Is Variable Rice bran (14) Min Mean Max Rice bran (14) Min Mean Max Wheat midds (8) Wheat midds (8) Canola (24) Canola (24) Cottonseed (6) Cottonseed (6) SBM (56) SBM (56) Corn (56) Corn (56) Wheat (28) Wheat (28) Sorghum (29) Sorghum (29) Barley (12) Barley (12) Corn DDGS (17) Corn DDGS (17) Rye (3) Rye (3) 0.00 2.00 4.00 6.00 8.00 % IP6 0.00 0.50 1.00 1.50 2.00 % Phytate P

Total AA digestibility (%) Phytate Degradation by Phytase Increases Protein Digestibility 95 90 R² = 0.67 85 80 75 70 65 60 20 30 40 50 60 70 80 90 100 Phytate degradation (%) There is a strong correlation between phytate breakdown and protein digestibility in vivo Amerah et al., 2012

Stomach Phytate Increases Costly Endogenous Secretions Protein de novo formation of protein-phytate complexes Phytate Refractory to pepsin digestion Additional outputs of pepsin and HCl ------------------------------------------------------------------------------------------------------------------------------------------------------------------ Small Intestine Extra mucin secretion Pre-bound dietary protein less readily digested Na + as NaHCO 3 Reduction of Na + - dependent transport and sodium pump activity Reduced absorption of dietary amino acids and reduced re-absorption of endogenous amino acids Mucin loss Source: Based on Selle, 2008 Compromised amino acid uptakes

A Look at the Future 20

Future Feed Stocks New Challenges Protein for poultry from grass Algae from ethanol plant shows promise as poultry feed http://ethanolproducer.com/articles/8220/algae-fromethanol-plant-shows-promise-as-poultry-feed 21

Biomass Challenge 1. Hemicellulose: broken down with exogenous enzymes eg. Xylanase 2. Cellulose: Crystalline structure, difficult for enzymes to access 3. Lignin: non-fermentable, no energetic value for animals Sannigrahi et al., 2010

Personalised Nutrition/Medicine http://www.hireanillustrator.com/i/49708/new-illustration-for-nature-magazinemedical-genomics/ DuPont Confidential 23

Summary Dupont is committed to Food Security Enzymes gives» Increased animal production efficiency» More sustainable production» Healthier better perfoming animals The future» New alternative non-food feedstocks» New technologies applied to animal production 24

Charlotte Horsmans Poulsen charlotte.poulsen@dupont.com Copyright 2013 DuPont. All rights reserved. 25

Acknowledgements Luis Romero Ajay Awati Gary Partridge Karsten M Kragh Steve Betz DuPont Confidential 26