Challenges of Predicting Metabolizable Lysine Content of...
Transcript of Challenges of Predicting Metabolizable Lysine Content of...
Challenges of Predicting Metabolizable Lysine Content of
Ingredients
Sarah BoucherWilliam H. Miner Agricultural Research Institute
Pre-Conference Symposium
71st Annual Cornell Nutrition ConferenceEast Syracuse, NY
October 20 – 22, 2009
Presentation
1. Lysine structure
2. Nutritional consequences of heat processing
3. Estimating lysine damage in the RUP fraction of feeds
4. Digestibility of lysine and RUP in blood meal
5. Summary
20 Amino acids
Indispensable 1. Arginine2. Histidine3. Isoleucine4. Leucine5. Lysine6. Methionine7. Phenylalanine8. Threonine9. Tryptophan10. Valine
Dispensable1. Alanine2. Aspartic acid3. Asparagine4. Cysteine5. Glutamic acid6. Glutamine7. Glycine8. Proline9. Serine10. Tyrosine
Protein metabolism in ruminantsCrude protein Saliva
True protein
Peptides
Aminoacids
Ammonia
NPN
Microbial protein
Urea
Liver
Microbial protein
RUMEN
SMALLINT
Endogenousprotein
Metabolizable protein (absorbed AA)
RUP
Mammarygland
MILK
Aminoacids
RUP
Characteristics of Lysine
Cationic amino acid at physiological pH In direct competition with arginine
for intestinal absorption Absorption stimulated by intracellular
leucine
- S. Bröer 2008. Physiol Rev 88:249-286
Heat Processing Reactions
1. Amino acid racemization
2. Protein cross-linking reactions
3. Maillard reaction
Maillard Reaction
Generally has the greatest impact on nutritional quality of feeds
3 phases:1. Early2. Advanced 3. Final
Simplified Scheme of Maillard Reaction
Adapted from J. Mauron. 1981. Prog. Fd. Nutr. Sci. 5:5-35.
Amadori compound
Melanoidin formation
Reducing sugar + Amino compound
Advanced glycation end products
Challenges with Lysine Analysis
H2N COOHCH
(CH2)4
NH
CH
(HCOH)3
C = O
CH2OHAmadori compound
Boiling 6N HCl
24 h
H2NCH
H2NCH
H2N COOHH2N COOHCH
H2N COOHCH
H2N COOHCH
H2N COOHCH
(CH2)4
COOHCH
(CH2)4
COOHCH
(CH2)4
COOHCH
NH
(CH2)4
COOHCH
NH
(CH2)4
COOHCH
CH
NH
(CH2)4
COOHCH
C = O
CH
NH
(CH2)4
COOHCH
(HCOH)3
C = O
CH
NH
(CH2)4
COOHCH
CH2OH
(HCOH)3
C = O
CH
NH
(CH2)4
COOHCH
Terminology
Bioavailability – proportion of ingested dietary AA that is absorbed in a chemical form that renders these AA potentially suitable for metabolism or protein synthesis
Digestibility – reflects enzymatic hydrolysis of ingested proteins and absorption of AA and peptides from the gastointestinal lumen
Terminology
Basal endogenous AA losses - the minimum quantities of AA inevitably lost by the animal
Standardized digestibility –calculated by subtracting only basal endogenous AA losses from outflow of AA
Standardized Digestibility –SBM Products
0
10
20
30
40
50
60
70
80
90
100
RUP-Lysine RUP-Total AA
SE = 2.9 SE = 1.7
Stan
dard
ized
dig
esitb
ility
, %
HSPHSBMSP2SP3SBM2SBM3
a,b,c Means within a series with different superscripts differ, P < 0.01
c
b
a a a a
c
b
a a a a
Standardized Digestibility -DDGS
0
10
20
30
40
50
60
70
80
90
100
RUP-Lysine RUP-Total AA
SE = 2.93 SE = 1.28
Sta
ndar
dize
d di
gest
ibilit
y, %
HDDGSDDGS1DDGS2DDGS3DDGS4
a,b,c Means within a series with different superscripts differ, P < 0.01
c
b
a a,b a,b
b
a a a a
Standardized Digestibility –Fish meals
0
10
20
30
40
50
60
70
80
90
100
RUPLysine RUP-Total AA
SE = 1.88 SE = 1.44
Stan
dard
ized
dig
estib
ility
, %
ANVYCFSHMNHN1MNHN2PLCK
a,b,c Means within a series with different superscripts differ, P < 0.01
a
b
a a a a
b
a a a
Terminology
Blocked lysine – lysine molecules in which the ε-amino group is bound to another compound
Reactive lysine – lysine molecules in which the ε-amino group is notbound to another compound
Estimating Blocked RUP-Lys
H2N COOHCH
(CH2)4
NH
CH
(HCOH)3
C = O
CH2OHfructosyllysine
Boiling 6N HCl
24 h
H2NCH
H2NCH
H2N COOHH2N COOHCH
H2N COOHCH
H2N COOHCH
H2N COOHCH
(CH2)4
COOHCH
(CH2)4
COOHCH
(CH2)4
COOHCH
NH
(CH2)4
COOHCH
NH
(CH2)4
COOHCH
CH
NH
(CH2)4
COOHCH
C = O
CH
NH
(CH2)4
COOHCH
(HCOH)3
C = O
CH
NH
(CH2)4
COOHCH
CH2OH
(HCOH)3
C = O
CH
NH
(CH2)4
COOHCH
Estimating Reactive RUP-Lys
NH2
CH2
CH2
CH2
CH2
C COOHH2N
H
Lysine
CNH
O
H2N
CH3
H
C
H
COOHC
H
H2N COOH
H
CH2N COOH
H
CH2
CH2N COOH
H
CH2
CH2
CH2N COOH
H
CH2
CH2
CH2
CH2N COOH
H
CH2
CH2
CH2
CH2
CH2N COOH
H
NH2
CH2
CH2
CH2
CH2
CH2N COOH
H
NHCH2
CH2
CH2
CH2
CH2N COOH
H
NH2
CH2
CH2
CH2
CH2
CH2N COOH
H
NH2
CH2
CH2
CH2
CH2
CH2N COOH
H
C NH2
NH
Homoarginine
Modified TSP Gargallo et al. (2006)
5 g of rumen undegraded resiude weighed into polyester bags in duplicate Incubated in a pepsin/HCl
solution 1 h, 38°C Incubated in a pancreatin
solution 24 h, 38°C
Bag residues analyzed for AAGargallo et al. (2006) J. Anim. Sci. 84:2163-2167.
Lysine Estimates – Soy-productsMethod
FurosineHomo-
arginine MTSP Rooster
Sample
Blocked RUP-Lys,
%
Reactive RUP-Lys,
%
RUP-Lys digestibility,
%
RUP-Lys digestibility,
%
Heated SP - 36.5 66.4 37.8
SP1 - 81.5 92.7 89.5
SP2 0.1 77.4 94.1 84.9
Heated SBM - 43.3 68.4 55.0
SBM1 0.7 85.2 97.7 90.1
SBM2 1.0 85.1 98.8 89.6
Lysine Estimates – DDGS
Method
FurosineHomo-
arginine MTSP Rooster
Sample
Blocked RUP-Lys,
%
Reactive RUP-Lys,
%
RUP-Lys digestibility,
%
RUP-Lys digestibility,
%
Heated DDGS - 31.5 32.4 10.3
DDGS2 26.0 70.3 79.2 63.0
DDGS3 7.6 76.8 92.1 79.5
DDGS4 15.3 73.7 86.7 75.8
DDGS5 21.0 71.3 89.1 72.7
Lysine Estimates – Fish meal
Method
FurosineHomo-
arginine MTSP Rooster
Sample
Blocked RUP-Lys,
%
Reactive RUP-Lys,
%
RUP-Lys digestibility,
%
RUP-Lys digestibility,
%
ANVY 0.4 84.9 95.9 87.5
CFSH - 71.3 66.9 63.2
MNHN1 0.2 79.5 95.4 88.6
MNHN2 - 79.1 92.6 84.3
PLCK - 89.4 92.0 89.9
Blood mealMethod
Homo-arginine
MTSP1 MTSP2 Rooster
Sample
Reactive RUP-Lys,
%
RUP-Lys digestibility,
%
RUP-Lys digestibility,
%
RUP-Lys digestibility,
%
Heated bovine BM 45.3 56.7 62.0 93.2
Bovine BM1 67.0 71.3 81.7 93.9
Bovine BM2 73.0 71.2 76.7 85.8
Heated porcine BM 86.8 17.2 19.5 83.9
Porcine BM1 71.3 93.4 80.4 92.0
Porcine BM2 93.1 - 94.7 95.3
Ohio State Modifications ofthe Three-Step Procedure
1. Foundation for the procedure is the Calsamiglia and Stern (1995) Minnesota three-step procedure
2. Modifications minimized inter –assay variation to 5%
3. Validated by Nostfsger and St-Pierre (2003, J. Dairy Sci. 86:958-969) lactational performance trial
4. Patent pending procedure is intellectual property of Venture Milling
Blood Meal RUP digestibility
Blood meal RUP digestibility
0102030405060708090
100110120
0 10 20 30 40 50 60 70 80 90 100 110 120
RUP (%CP)
RUP
diges
tibilit
y %
Data courtesy of Venture Milling. Ring dried blood meal data generated from non-published Ohio State University research using a Modified Minnesota 3-Way analytical procedure.
n = 403
Average RUP digestibility, % = 64.6; SE = 23.1
Blood Meal RUP-Lysine Digestibility
Blood meal RUP lysine digestibility vs RUP digestibility
0102030405060708090
100
0 10 20 30 40 50 60 70 80 90 100
RUP digestibility
RUP
Lys
dig
estib
ility
Data courtesy of Venture Milling. Ring dried blood meal data generated from non-published Ohio State University research using a Modified Minnesota 3-Way analytical procedure.
n = 403
Average RUP- Lys digestibility, % = 56.0; SE = 27.1
NRC Crude Protein Fractions
Feed Crude Protein
Fraction CUndegraded
Fraction BRest of CP
Fraction ANPN
NRC, 2001
RUP digestibility, NRC (2001)
RUP digestibility values are the approximate mean values reported using the mobile bag technique and the three step procedure
Assumes digestibility of individual AA in RUP is the same as RUP
CNCPS Crude Protein Fractions
TOTAL
BORATEBUFFER
NEUTRALDETERGENT
ACIDDETERGENT
SOLA
B1
INSOL
B2B3
C
SOLA
B1B2
INSOL
B3
C
SOLA
B1B2B3
INSOL
C
RUP digestibility, CNCPS v.6.1
Absorption of escaped feed protein is calculated by multiplying each protein fraction by its respective intestinal digestibility A – 100% B1 – 100% B2 – 100% B3 – 80% C – 0%
Assumes digestibility of individual AA in RUP is the same as RUP
Ingredient % of DMCorn silage 36.8Grass silage 16.1Alfalfa hay 5.8Corn grain 10.7Soybean hulls 5.0Soybean meal 2.1DDGS 10.7Soybean meal, expellers 2.3Blood meal 1.3Rumen protected Met 0.05Urea 0.25Inert fat 1.1Vitamins/minerals 2.5
Example diet
Balanced in NRC (2001)
Animal:
~ 85 lbs milk
DMI = 53 lbs
NRC (2001) Library
DDGS Blood mealring-dried
RUP, % CP* 50 78
RUP digestibility, % 80 80
Lys, % CP 2.7 8.9
*At DMI = 4.0% of BW and DMI = 50% forage
RUP-Lys digestibility
Scenario 1: DDGS RUP-Lys digestibility = 63% Blood meal RUP-Lys digestibility = 56%
Scenario 2: DDGS RUP-Lys digestibility = 80% Blood meal RUP-Lys digestibility = 90%
Example
NRC (2001) Scenario 1 Scenario 2
Lys, % of MP 6.22 5.99 6.30
Met, % of MP 2.07 2.07 2.07
Lys: Met 3.0:1 2.89:1 3.04:1
Summary
Heat processing of feeds can damage lysine
Intestinal digestibility of lysine in the RUP fraction of heated feeds is less than digestibility of total RUP
Determining digestible or available lysine content of feeds can improve predictions of MP-Lys supply
Acknowledgements
Special thanks to Venture Milling and Normand St-Pierre
Financial support for some of the research presented was provided by: Adisseo West Central
Special thanks to research collaborators Carl Parsons Pam Utterback Hans Stein Sergio Calsamiglia Marshall Stern Carsten Pedersen Lawrence Novtony &
Deon Simon at SDSU