Mr Jean-Paul Ruckebusch

Transcription

1 Mr Jean-Paul Ruckebusch Global Category Manager Enzymes, DSM Nutritional Products Ltd How to overcome anti-nutritional factors by using a range of enzymes Sponsored by:

2 How to overcome anti-nutritional factors by using a range of enzymes Global Feed and Food Congress 2013 South-Africa April 10, 2013 J-P. Ruckebusch, I. Knap, M. Umar Faruk and E. Upton Augustsson

3 CONFIDENTIAL Public/How to overcome anti-nutritional factors with a range of feed enzymes/j-p Ruckebusch/April, 2013 Anti-Nutritional Factors & Enzymes Plants produce ANFs as part of their normal metabolism ANFs interfere with animal s ability to digest plant nutrients, thereby reducing digestive efficiency by Binding to various nutrients Increasing viscosity of intestinal contents Inhibiting/deactivating endogenous digestive enzymes Irritating the intestinal mucosa (challenge to immune system) Specific mono-component enzymes work against ANFs by hydrolyzing: Phytin Non-soluble and soluble NSPs Trypsin inhibitors and lectin Other ANFs (ß-mannan, etc ) Page 2 Source: Kumar, 1994; Choct et al., 1991; Lange et al., 2000; Cowieson 2004

4 Phytate life cycle Proteins Proteins Proteins Phytate in plant cell wall Proteins Phytate negative charges bind various positively charged molecules Insoluble binary protein-phytate complexes Starch Proteins Proteins Starch Starch Insoluble tertiary protein-mineral-phytate complexes with Ca Mineral-phytate complexes Page 3 Source: Rajendran and Prakash, 1993; Kies et al., 2006; Prattley et al., 1982; Montagne et al., 2001

5 Phytate & Phytase in vivo O Cu ++ - O O - O - O O - P Ca ++ P Fe ++ - O O P O O O - CH CH H C O O C H K + - O HC HC P O - O O Mg ++ - O O O P P O Na O O - O Zn ++ Hydrolyses IP6 into IP3 Liberates P, Ca, Zn, Reduces protein binding Various soluble IP3 are hydrolyzed by endogenous enzymes - Available P - Available Ca - Available minerals Less excretion Bone strength and mineralisation Physiologic functions Growth performances - Available amino acids* Page 4 Source: Mroz, Jongbloed et al., 1994; Rapp et al., 2001; Kemme et al., 2006 *48 PR papers, 34 in poultry, 80% positive effects on AA digestibility

6 Phytate w Phytase in vivo In presence of phytase, phytates (IP6) are hydrolyzed in smaller phytate esters like IP4, IP3 Small phytate esters are soluble and do not bind proteins. They will be degraded and absorbed Not bound proteins are available for hydrolysis into peptides and amino acids By indirect action phytase increases nutrient digestibility Page 6

7 Model of plant cell wall A mixture of polysaccharides interlocked in a complex structure Xyloglucan network Cellulose fibrils Cellulose fibrils Glucuronoarabinoxylan Monocotyledonous plants (like corn, wheat, barley) Pectic polymer matrix Dicotyledonous plants (like soya, canola, sunflower) Page 7 Source: Carpita, N.C. and Gibeaut, D. M (1993).The Plant Journal 3(1):1-30.

8 Different plant cell walls carbohydrates Different plant cell wall soluble and insoluble carbohydrates require different enzymes solutions Page 8 Source: Bach Knudsen KE., 1997; Choct. M.,1997

9 The insoluble cell wall structure in corn Endosperm cell wall w. insoluble arabinoxylans Broken down by xylanase The thin cell walls can be described as arabinoxylan cages encapsulating nutrients. Releasing cell contents Page 9 Source: Evers and Millar 2002

10 Degradation of cell walls by xylanase 0 min 180 min Wheat cuts right after injection of xylanase Wheat cuts 180 minutes after treatment with xylanase Fluorescence is induced by UV irradiation of plant materials, then emitted by mainly ferulic acid cross-linking arabinoxylan chains. Page 10 Source: Le et al., 2013

11 Degradation of cell walls by xylanase 0 min 120 min Corn from piglets digesta sample right after injection of xylanase Corn from piglets digesta sample 120 minutes after treatment with xylanase The degradation of cell walls by a xylanase is monitored indirectly as a decrease in fluorescence resulting from the breakdown of the arabinoxylan rich cell walls into soluble oligomers. Page 11 Source: Novozymes, unpublished

12 Public/How to overcome anti-nutritional factors with a range of feed enzymes/j-p Ruckebusch/April, 2013 Degradation of cell walls by pure xylanase Xylanase hydrolyses the non-soluble fraction of plant cell walls Releases the entrapped nutrients Opens the cell wall structure Give access to digestive enzymes to cell content Xylanase hydrolyses the soluble fraction of plant cell walls Increases Feed Intake Decreases intestinal viscosity Increases passage rate of digesta Reduces substrate for negative microflora Xylanase Xylanase Insoluble arabinoxylans Viscous soluble arabinoxylans Non viscous soluble arabinoxylans Page 12

13 Weight gain (g/bird/week) FCR Different mode of action of pure xylanases The in vivo confirmation ,156 Active on both soluble and insoluble arabinoxylans Active only on insoluble arabinoxylans Active only on soluble arabinoxylans 2,2 2, , ,965 2,005 2,035 2, , ,4 392,2 394,8 382,9 Control Xylanase Exp. xylanase Test product Best performances require xylanase activity against both insoluble and soluble xylans. 1,9 1,85 Page 13 Source: Choct, M., Kocher, A., Waters, D.L.E., Pettersson, D. and Ross, G., 2004

14 CONFIDENTIAL Feed Protease degrades soya ANFs Lectin Kunitz Inhibitor Bowman Birk Inhibitor Degradation % Protease Trypsin Lectin Kunitz 96 4 Bowman Birk Page 14 Source: Nielsen et al., 2013

15 Protein hydrolysis - ABS 340 nm CONFIDENTIAL ANFs in soy and protease s efficacy Trypsin Feed Protease Pancreas Trypsin Novo, g/kg SBM Feed Protease, 1000 Units/kg SBM Raw soy SBM 1 SBM 2 Feed Protease acts by reducing Lectin level and KTI and BBI levels, thereby reducing the sensitivity of animal performance to low quality soybean meal. Page 15 Source: Nielsen et al., 2013

16 Protein hydrolysis - ABS 340 nm CONFIDENTIAL ANFs in soy and protease s efficacy Pancreatic Enzymes + Feed Protease Pancreatic Enzymes Feed Protease Commercial dose Pancreas Trypsin Novo, g/kg SBM The addition of the pure feed protease completes and doubles the efficacy of endogenous pancreatic proteases (trypsin and chymotrypsin). Page 16 Source: Nielsen et al., 2013

17 Body weight gain (g/b) CONFIDENTIAL Raw soybean & protease on broilers % (P<0.05) % (P>0.05) % (P<0.05) % RSB 10% RSB 20% RSB Page 17 Source: Umar Faruk et al., 2012

18 DH relative to no protease % % P-release from InsP 6 reletive to phytase alone (%) CONFIDENTIAL Feed Protease combined with phytases Degree of protein hydrolysis +/- protease % n=4 * n= P-release from phytate (IP 6 ) +/- protease 0 No protease Protease * Significantly different from control (All-pairs Tukey Kramer HSD, P<0.05) 50 Overall significant positive effect of phytase+protease compared to phytase alone on phytate degradation Page 18 Source: Pontoppidan et al., 2012

19 DH relative to no protease % Soluble xylose relative to xylanase alone % (%) CONFIDENTIAL Feed Protease combined with xylanases Degree of protein hydrolysis +/- protease % * n = Xylose solubilization +/- protease 50 n = No protease Protease * Significantly different from control (All-pairs Tukey Kramer HSD, P<0.05) 0 Overall significant positive effect of xylanase+protease compared to xylanase alone on xylose degradation Page 19 Source: Pontoppidan et al., 2012

20 CONFIDENTIAL One Substrate, One enzyme Studies on ANFs in feed ingredients on which mono-component enzymes have proven to be effective provide a better understanding of the mode of action of specific feed enzymes. It also helps in predicting the nutritional contribution a single enzyme or a combination of enzymes can make to the value of a diet. Enzymes are highly specific. The non-soluble and soluble NSPs are more complex substrates and may require more than one enzyme to alleviate the ANF s impact. The choice of commercial feed enzymes and combination must be made based on the best knowledge available, not only on the enzyme activity itself, but on the substrate it is targeting. Page 20

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