The research shows the apparent and the true amino acid digestibility of full fat soy with and without protease (RONOZYME ProAct) supplementation. Join the Feed Enzyme discussion on Linked in: http://www.linkedin.com/groups?home=&gid=4738175&trk=anet_ug_hm&goback=%25 Follow us on Twitter: @DSMFeedTweet Or speak to an expert: @Jobsorbara

1. EFFECT OF A MONO COMPONENT PROTEASE ON
TRUE AMINO ACID DIGESTIBILITY OF FULL FAT SOY
FOR BROILER CHICKENS USING DIFFERENT
METHODS.
R. K. G. Messias1, L. F. T. Albino1, J. O. B. Sorbara2*, H. S.
Rostagno1
Universidade Federal de Viçosa - Brazil
2
DSM Nutritional Products - Brazil
1

2. Introduction
• Poultry Science Meeting 2009
– Several abstracts presented
Improvement (% Control)
•Protease to enhance the amino acid utilization of different raw ingredients by broilers.
115%
110%
105%
100%
Asp Glu Ser
Corn
Gly
His Arg Thr
Soy Bean Meal
Ala Pro
Tyr
Full Fat Soy
Val Met Cys
Ile
Meat Bone Meal
Leu Phe Lys

3. Introduction
• Poultry Science Meeting 2009
– Several abstracts presented
•Protease to enhance the amino acid utilization of different raw ingredients by broilers.
– Conclusions:
• The mono-component protease used was enable to improve de AA digestibility of
different ingredients between 2 and 15% dependent of individual AA.
– Discussion:
• The response was based on Total Apparent AA digestibility
• Today the raw materials are evaluated by True Ileal AA digestibility
• Would the response of this mono-component protease be different if
the True Ileal AA digestibility method was used?
• How different methods can effect the enzyme response?

4. Objectives
• Compare the Apparent and True Ileal Amino Acid
Digestibility of the Full Fat Soy with and without the
protease supplementation.
• Compare the effect of different methods on the Full Fat
Soy Amino Acid Digestibility in broiler chickens.

5. Materials & Methods
• Experimental Period
• Metabolism Trial
– 336 Male Cobb
– Experimental Design
• 8 treatments
• 6 replicates of 7 birds each
Tr.
– 12 to 18 d old adaptation period
– 19 to 21 d old total excreta
collection
– At 22 d old euthanized by cervical
dislocation to collect the ileal
digesta
Tr.
Without Protease
With Protease
1
Protein Free Diet (PFD)
5
Protein Free Diet
2
Protein Free Diet + Full Fat Soy
6
Protein Free Diet + Full Fat
Soy
3
Basal Diet + Starch
7
Basal Diet + Starch
4
Basal Diet + Full Fat Soy
8
Basal Diet + Full Fat Soy

6. Materials & Methods
• Diets Composition
Ingredients/ Diets
Starch
Full-fat soy (FFS)
Basal diet
Sugar
Corn
Soybean meal
Soybean oil
Dicalcium fosfate
Limestone
Salt
Corn cob
L-lysine HCl 99%
DL-methionine, 99%
L-threonine, 98%
Mineral premix1
Vitamin premix2
Choline chloride
Antioxidant (BHT)
Acid-insoluble ash (CeliteTM)
Total
Crude protein
PFD
78.01
5.00
5.00
2.10
1.00
0.45
4.00
0.08
0.15
0.20
0.01
1.00
100.0
0.00
PFD + FFS
48.01
30.00
5.00
5.00
2.10
1.00
0.45
4.00
0.08
0.15
0.20
0.01
1.00
100.0
11.10
Basal + Starch Basal + FFS
30.00
30.00
70.00
70.00
100.0
100.0
14.70
25.80
Basal
57.13
36.00
2.00
1.85
0.90
0.45
0.11
0.23
0.04
0.05
0.10
0.10
0.01
1.00
100.0
21.00

7. Materials & Methods
• Full Fat Soy Composition
(analyzed)
Nutrient
%
CP
35.95
Fat
19.1
Solubility in KOH
77.87
Urease, %
0.04
Energy, Kcal
5168
DM, %
88.31
Amino Acids (%)
Aspartic Acid
Glutamic Acid
Ser+Gly
Histidine
Arginine
Threonine
Alanine
Proline
Tyrosine
Valine
Methionine
Cystine
Isoleucine
Leucine
Phenylalanine
Lysine
Total amino acids
FFS 2010
4.04
6.61
3.78
0.98
2.94
1.5
1.57
2.05
1.44
1.51
0.55
0.4
1.57
2.9
2
2.11
35.92

8. Materials & Methods
Comparative
Full Fat Soy Composition
• Bertechini et al., (2009)
• Messias et al., (2010)
Amino Acids (%)
Aspartic Acid
Glutamic Acid
Ser+Gly
Histidine
Arginine
Threonine
Alanine
Proline
Tyrosine
Valine
Methionine
Cystine
Isoleucine
Leucine
Phenylalanine
Lysine
Total amino acids
FFS
2009
4.01
6.23
3.29
0.91
2.58
1.27
1.53
1.69
1.43
1.61
0.62
0.56
1.58
2.67
1.75
2.41
34.51
FFS
2010
4.04
6.61
3.78
0.98
2.94
1.5
1.57
2.05
1.44
1.51
0.55
0.4
1.57
2.9
2
2.11
35.92
Dif (%)
0.7%
6.1%
14.9%
7.7%
14.0%
18.1%
2.6%
21.3%
0.7%
-6.2%
-11.3%
-28.6%
-0.6%
8.6%
14.3%
-12.4%
4.1%

9. Materials & Methods
• Analyses
– Samples were freeze-dried
– Dry Matter
– Crude Protein
– Acid-Insoluble Ash (AIA)
– Amino Acids analyzed by HPLC
• Calculations:
Indigestility Factor (IF)
– IF1= [AIA] in the test diet
[AIA] in the sample
– IF2 =
[AIA] in PFD
[AIA] in the sample
Ileal Digestibility Coefficient (IDC)
Apparent IDC = (AA% in the diet – [amino acid % in the digesta x IF1]) x 100
AA % in the diet
True IDC = (%AA in the diet – ([%AA in the digesta x IF1] – [endogenous AA x IF2]) x 100
AA % in the diet

10. Materials & Methods
• Product Specifications
– Mono component protease with 75000 PROT/g originated from
Nocardiopsis prasina and produced by the bacteria Bacillus
licheniformis (RONOZYME ProAct, DSM)
– Dose 15000 PROT/kg = 200 ppm
– One ProAct activity unit (PROT) is the amount of enzyme that
releases 1 µmol of p-nitroaniline from 1 mM substrate (Suc-AlaAla-Pro-Phe-pNA) per minute at pH 9.0 and 37 °C.

11. Results
• Effect of Mono Component Protease on Full Fat Soy
True Ileal AA digestibility coefficient.
Protease
Without
With
ANOVA
CV(%)
Improvement (% Control)
Asp
79,1
88,5
**
2,6
112
Glu
83,3
90,3
**
2,2
108
Ala
80,5
86,3
**
4,2
107
Pro
81,9
88,5
**
3,1
108
Tyr
82,9
89,4
**
3,1
108
Cys
70,3
81
**
8,3
115

12. Results
• Effect of Mono Component Protease on Full Fat Soy
True Ileal AA digestibility coefficient.
Protease
Without
With
ANOVA
CV(%)
Improvement (% Control)
Val
83,8
85,5
ns
4,3
102
His
81,5
88,5
**
2,6
107
Arg
88,2
92,1
**
1,8
104
Ile
84,6
86,9
0,08
3,6
103
Leu
87,4
86,9
ns
3,6
99
Phe
83,6
88,0
**
2,9
105

13. Results
• Effect of Mono Component Protease on Full Fat Soy
True Ileal AA digestibility coefficient.
Protease
Without
With
ANOVA
CV(%)
Improvement (% Control)
Lys
86,4
89
**
2,3
103
Met
89,4
92,4
**
2,6
103
Met+Cys
79,9
86,7
**
5,4
109
Thr
78,4
83,9
**
5,3
107
Sum
82,3
90,1
**
2,7
109
CP
87,1
90,3
**
3,7
104

14. Results
• Effect of Method on Full Fat Soy True Ileal AA digestibility
coefficient.
Protease
PFD
Basal Diet
ANOVA
CV(%)
Dif (BD/PFD x 100)
1
Asp1
93,6
74,0
**
2,6
79
Glu1
95,7
78,0
**
2,2
82
Enzyme vs. Method Interaction (P< 0,05)
Ala
92,7
74,1
**
4,2
80
Pro
95,5
74,9
**
3,1
78
Tyr
94,8
77,4
**
3,1
82
Cys
89,7
61,6
**
8,3
69

15. Results
• Effect of Method on Full Fat Soy True Ileal AA digestibility
coefficient.
Protease
PFD
Basal Diet
ANOVA
CV(%)
Dif (BD/PFD x 100)
1
Val
93,9
75,4
**
4,3
80
His
95,1
75
**
2,6
79
Enzyme vs. Method Interaction (P< 0,05)
Arg 1
97,4
83
**
1,8
85
Ile
94
77,5
**
3,6
82
Leu
95,4
78,9
**
3,6
83
Phe
95,2
76,3
**
2,9
80

16. Results
• Effect of Method on Full Fat Soy True Ileal AA digestibility
coefficient.
Protease
PFD
Basal Diet
ANOVA
CV(%)
Dif (BD/PFD x 100)
Lys
94,6
80,8
**
2,3
85
Met
96
85,8
**
2,6
89
Met+Cys Thr
92,9
90,2
73,7
72
**
**
5,4
5,3
79
80
Sum
95,1
77,2
**
2,7
81
CP
92,9
84,6
**
3,7
91

17. Results
• Overall response of mono component protease on the Full Fat Soy
Digestibility by different AA digestibility methods and Apparent
and True Ileal AA digestibility coefficient
Apparent AA Dig using BD
True AA Dig using PFD Method
True AA Dig using BD
120%
115%
110%
105%
Le
u
Ph
e
Ly
s
Ile
100%
A
sp
G
lu
Se
r
G
ly
H
is
A
rg
Th
r
A
la
Pr
o
Ty
r
Va
l
M
+C
Improvement (% Control)
Apparent AA Dig using PFD Method

18. Results
• Overall response of mono component protease on the Full Fat Soy
Digestibility by different AA digestibility methods and Apparent
and True Ileal AA digestibility coefficient
Apparent AA Dig using BD
True AA Dig using PFD Method
Improvement (% Control)
Apparent AA Dig using PFD Method
True AA Dig using BD
120%
115%
110%
105%
100%
His
Arg
Thr
Val
M+C
Ile
Leu
Phe
Lys

19. Discussion
• Comparing responses from Prof. Bertechini et al., (2009)
and Messias et al., (2010)
2009 - Apparent AA Dig using BD
2010 - Apparent AA Dig using PFD Method
2010 - Apparent AA Dig using BD
2010 - True AA Dig using PFD Method
120%
115%
110%
105%
Le
u
Ph
e
Ly
s
Ile
100%
As
p
G
lu
Se
r
G
ly
Hi
s
Ar
g
Th
r
Al
a
Pr
o
Ty
r
Va
l
M
+C
Improvement (% Control)
2010 - True AA Dig using BD

20. Discussion
• Comparing responses from Prof. Bertechini et al., (2009)
and Messias et al., (2010)
2009 - Apparent AA Dig using BD
2010 - Apparent AA Dig using PFD Method
2010 - Apparent AA Dig using BD
2010 - True AA Dig using PFD Method
Improvement (% Control)
2010 - True AA Dig using BD
120%
115%
110%
105%
100%
His
Arg
Thr
Val
M+C
Ile
Leu
Phe
Lys

21. Conclusions
• These methods did not affect the protease response on
Full Fat Soy amino acid digestibility.
• Even when using different methods or Apparent vs True
amino acid digestibility the protease response on full fat
soy was similar.

22. Thank you for your attention !!!

23. Back-up

24. Results
• Effect of Mono Component Protease on Full Fat Soy
Apparent Ileal AA digestibility coefficient.
Protease
Without
With
ANOVA
CV(%)
Improvement (% Control)
Asp
76,4
84,5
**
2,6
111
Glu
79
84,6
**
2,3
107
Ala
71,5
75,6
**
4,5
106
Pro
71,4
76,4
**
3,3
107
Tyr
76,4
78,5
0,07
3,4
103
Cys
48,8
59,5
**
10,3
122

25. Results
• Effect of Mono Component Protease on Full Fat Soy
Apparent Ileal AA digestibility coefficient.
Protease
Without
With
ANOVA
CV(%)
Improvement (% Control)
Val
70
73,4
0,02
4,7
105
His
76,8
81,5
**
2,7
106
Arg
81,9
86
**
1,9
105
Ile
76,9
78,4
ns
3,8
102
Leu
80,4
79,5
ns
3,8
99
Phe
75,9
79,9
**
3,1
105

26. Results
• Effect of Mono Component Protease on Full Fat Soy
Apparent Ileal AA digestibility coefficient.
Protease
Without
With
ANOVA
CV(%)
Improvement (% Control)
Lys
82,5
84,0
0,086
2,34
102
Met
80,7
85,0
**
2,7
105
Met+Cys
64,7
72,2
**
6,5
112
Thr
63,2
69,8
**
6,0
110
Sum
74,8
79,6
**
3,0
106
CP
80,7
83,0
0,11
4,0
103

27. Visit us on:
•DSM Animal Nutrition and H