Proteases in Feed!
Jefo Poultry and Swine Tour! China Jacobs, Ph.D!
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Proteases in Feed! OUTLINE!
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1.
What it is and why we might use one!
2.
Key factors to consider when choosing a protease!
3.
What does the bird say!
4.
Economic value!
5.
Conclusions!
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Background! INCREASING INTEREST IN FEED ENZYMES! !
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Source: Novus International!
Protease- What is it? ! • Definition: Enzymes that catalyze the hydrolysis (breaking) of peptide bonds between amino acids in proteins and polypeptides ! • Protein is a highly diverse substrate - 20 different amino acids to choose from at each position on a protein molecule!
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Protease Types ! Proteases!
Site of Action!
Nature of Active Site Residues!
Chymotrypsin!
Typsin-like!
5!
Elastaselike!
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Serine!
Metallo!
Exo-!
Endo-!
Cysteine!
Aspartic!
Subtilisin!
Chymotrypsinlike!
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Serine Protease Specificity ! • Specificity is largely determined by the amino acid residues present in the active site of the protease!
Lys! 6!
Arg!
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Ala!
Val!
Gly!
Tyr!
Phe!
Trp! !
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Adapted from Glisto and Pontoppidan, 2014!
Protease Specificity! Different proteases result in different hydrolysis products!
Very Specific Protease (Few Cuts)! ! Trypsin – only cleaves Arg and Lys!!
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Less Specific Protease (Many Cuts)! ! Chymotrypsin –preference for hydrophobic or aromatic amino acids !
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Adapted from Glisto and Pontoppidan, 2014!
AA digestibility is not optimal! !
Amino acids reaching ileum without digestion in presence of endogenous enzymes = substrate for exogenous protease and opportunity for economic benefit. ! ! 8!
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Source: Smith and Korsbak, 2014!
Amino acid profile of endogenous losses!
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Coweison, 2014!
Key Questions When Choosing a Protease ! Improvement (% Control)
1) Is it a broad spectrum protease? ! 115%
110%
105%
100% Lys
Met
Cys
Thr
Val
Corn Soy Bean Meal Full Fat Soy Corn DDGS Meat & Bone Meal Poultry Meal Jefo Poultry and Swine Tour! 10!
Ile
Arg
Canola Meal Feather Meal !
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Source: DSM!
Key Questions When Choosing a Protease ! 2) Is it stable throughout the feed manufacturing process (temp)? ! Amino Acids Hydrolyzed by Protease! Mash!
Pelleted (85°C)!
Extruded (95°C)!
(n=3)!
(n=3)!
(n=3)!
mg/mL-1!
21.84 ± 0.32a!
20.92 ± 0.44ab!
20.15 ± 0.45b!
% activity!
100!
95.7!
92.2! Khusro, 2016!
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Key Questions When Choosing a Protease ! 3) Does it work best in temperatures that match animal temperature (35-55°C)?! !
Amino acids hydrolyzed by protease at different reaction temps! 12!
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Key Questions When Choosing a Protease ! 4) Will it survive the low pH in the birds gut and work optimally in the small intestine? !
Khusro, 2016!
Source: Angel, 2013!
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Key Questions When Choosing a Protease ! 5) Is there evidence it will work in synergy with endogenous proteases or other enzymes? ! ! Effect of poultry specific protease on digestive enzymes and nutrient digestibility! Parameter!
Normal Diet! Low Density (CP + AA)! Normal Diet + Protease! Low Density + Protease!
Pepsin (AU)!
Amylase(AU)!
DM Digest (%)!
N Digest (%)!
97.23 ± 19.38a!
321.64 ± 40.07a!
70.67± 1.83ab!
77.54 ± 1.14a!
107.09 ± 14.25a!
68.09 ± 2.56a!
75.30 ± 2.17a!
155.61 ± 15.47b!
318.69 ± 25.43a! ! 380.87 ± 36.54a!
72.96 ± 3.27b!
79.67 ± 2.30b!
136.16 ± 15.37b!
326.14 ± 37.65a!
73.46 ± 2.62b!
79.97 ± 1.89b! Jefo, 2008!
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Key Questions When Choosing a Protease ! 5) Is there evidence it will work in synergy with endogenous proteases or other enzymes? ! 0-19 d! ! 600! 1.60! 580! 560!
1.55!
[VALEUR]a! [VALEUR]a!
540! 520!
[VALEUR]bc!
[VALEUR]ab! [VALEUR]a! [VALEUR]bc!
[VALEUR]ab!
1.40! [VALEUR]b!
480! 460!
1.45!
[VALEUR]c!
500!
[VALEUR]c!
1.35! 1.30!
440!
1.25!
420! 400!
1.20! PC!
NC!
NC + XYL! BWG!
15!
1.50!
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NC + PRT!
NC + XYL/ PRT!
FCR!
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Jefo, 2014!
What Does the Bird Say? !
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Improvement in AA Digestibility ! 100! 95!
100! Protease Improvement!
No Protease!
95!
+[VALEUR]!
75!
+[VALEUR]!
70! 65!
83.7!
81.9!
+[VALEUR]! 78.7!
60!
72.7!
62.5!
55! Corn!
DDGS!
MBM!
75! 70!
Bakery!
84!
83.1!
79.3!
76.9!
SBM!
100!
75.2!
Corn!
DDGS!
MBM!
Bakery!
100! Protease Improvement! +[VALEUR]!
No Protease! +[VALEUR]!
Protease Improvement!
No Protease!
90!
80! +[VALEUR]!
+[VALEUR]!
70! 60!
95!
90.2!
86.6! 74.9!
73.5!
+[VALEUR]!
Digestible Thr %!
Digestible Cys %!
+[VALEUR]! +[VALEUR]! +[VALEUR]!
80!
60! SBM!
50!
+[VALEUR]! 85! +[VALEUR]!
65!
50!
90!
No Protease!
90!
85! +[VALEUR]! +[VALEUR]! 80!
Digestible Met %!
Digestible Lys %!
90!
Protease Improvement!
85!
+[VALEUR]!
+[VALEUR]!
80! +[VALEUR]!
+[VALEUR]!
75! 80.6! +[VALEUR]!
70! 75.2!
40!
65!
42.6!
17! 30!
84.2! 76.8!
68.1!
60! SBM!
Corn!
DDGS!
MBM!
Bakery!
SBM!
Corn!
DDGS!
MBM!
Bakery!
Reduced endogenous losses!
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Angel, 2014!
What Does the Bird Say? ! CTRL 95%: formulated at 95% of the digestible lysine and CP levels of the control diet! CTRL 92%: formulated at 92% of the digestible lysine and CP levels of the control diet! Diets were corn-soy-DDGS based and fed from 1-42 days!
2.45!
BWG (kg)!
2.40!
[VALEUR]a!
[VALEUR]a!
[VALEUR]ab!
2.35! [VALEUR]bc!
2.30!
[VALEUR]c!
2.25! 2.20! 2.15! 2.10! CONTROL!
CRTL 95%! CTRL 95% + CTRL 92%! CTRL 92% + PROTEASE! PROTEASE!
1.78! 1.76!
[VALEUR]d!
FCR!
1.74!
[VALEUR]bc!
1.72!
[VALEUR]bc! [VALEUR]ab!
1.70! 1.68!
[VALEUR]a!
1.66! 1.64! CONTROL!
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CRTL 95%! CTRL 95% + CTRL 92%! CTRL 92% + PROTEASE! PROTEASE!
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Source: Jefo internal data, 2011!
What Does the Bird Say, Especially in Field Conditions? !
Integrated broiler company - 2520 chickens! Pelleted diets (80°C) with high inclusion of by-products, xylanase, and phytase !
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Jefo, 2012!
What Does the Bird Say, Especially in Field Conditions? !
Normal Protein!
• • • • 21!
Low Protein !
LP + Protease!
1100 Ross 708 males! Pelleted diets which included 5% DDGS and phytase ! Normal conditions: 21 bird/pen (14.3/m²)no feed outage ! Stress conditions: 25 birds/pen (17.2/m²), 8 hr feed outage at d0 and d14 !
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Economic Value! Broiler Weight: 2.72 kg! CA $/MT!
! Without Protease!
With Protease (AA)!
With Protease (ME ! & AA)!
Diet $/kg!
$/Phase!
Diet $/kg!
$/Phase!
Diet $/kg!
$/Phase!
Prestarter!
0.358!
0.058!
0.355!
0.057!
0.350!
0.056!
Starter!
0.346!
0.158!
0.343!
0.157!
0.337!
0.154!
Grower!
0.329!
0.526!
0.326!
0.521!
0.320!
0.511!
Finisher!
0.310!
0.759!
0.307!
0.752!
0.302!
0.740!
Total $/Broiler!
1.501!
1.487!
1.461!
Savings using Protease (AA) = 0.014 $/broiler! Savings using Protease (ME& AA) = 0.04 $/broiler!
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Conclusions! • Protease diversity exists (types and activity)! • For feed proteases, it is important to ensure that the key characteristics are in place:! • Ability to work over a wide range of feedstuffs! • Stability in the gut and during processing! • Synergy with endogenous enzymes! • Compatibility with other feed enzymes! • Most importantly - proven animal performance! ! • Proteases can generate feed cost savings, but need to test in your own system!
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