the production of bioactive peptides from peanuts -...
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FOOD Product Innovation and CommercializationFOOD Product Innovation and Commercialization
The Production of Bioactive Peptides from Peanuts
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Fat
Protein
Carbohydrate
Water
Other
Composition of peanut kernel
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Protein Structure
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Peptide Hydrolysis
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Peptides
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Amino Acid
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27
48
70
41
27
99
31
13
58
27
47
95
78
33
102
44
14
64
0 20 40 60 80 100 120
Histidine
Isoleucine
Leucine
Lysine
Methionine and cystine
Phenylalanine and tyrosine
Threonine
Tryptophan
Valine
Peanut mg/g Cows Milk
Essential Amino Acids
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Peptide structure
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Bioactive Peptides
• Bioactive Peptides –Those with physiological effect
• Antihypertensive (ACE inhibitor), antimicrobial, antithrombotic, antioxidant, etc.
• Various sources– Milk– Plant sources including legumes (chickpea)
• None reported from peanut
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S t r e s s
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S t r e s s
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Up Regulation of BP by ACE
Blood Pressure
Angiotensin I Angiotensin II
* Angiotensin I Converting Enzyme
ACE*
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Down Regulation of BP by ACE InhibitorsBlood Pressure
Angiotensin I Angiotensin II
* Angiotensin I Converting Enzyme
ACE*
ACE INHIBITORS
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OBJECTIVES
• To identify and isolate hypotensive peptides from proteolytic peanut digests
• To detect and evaluate bactericidal peptides from proteolytic peanut digests
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Methodology• Peanut Flour
– Digested with Alcalase & Pepsin-Pancreatin at different times using modified pH Stat
– Degree of hydrolysis using TNBS• Lyophilized Digest• ACE Activity• Antimicrobial
– E. coli & L. monocytogenes• SDS PAGE• HPLC
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METHODOLOGY Con’t
• HPLC on hydrolysates with highest DH values, from both raw and roasted peanuts digested with either enzyme– Fractions collected and ACE inhibition
determined• Antimicrobial activity also determined
using hydrolysates with the highest DH values
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Isolation of BiopeptidesCrude Hydrolysate
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METHODOLOGY Con’t
• Proteolytic digests tested against – Angiotensin Converting Enzyme (ACE) for
antihypertensive activity – E.coli O157:H7 and L. monocytogenes for
antimicrobial activity
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RESULTS
• Statistical analyses reveal– Degree of Hydrolysis
• Enzyme type and duration of hydrolysis significantly affect DH.
– Alcalase recorded higher DH values
• Peanut treatment has no significant effect on DH
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RESULTS Con’t
– ACE Activity• Hydrolysis time, Enzyme type and Peanut treatment
have a significant effect on ACE activity.– Raw peanuts recorded more hypotensive activity, and
hydrolysates from Alcalase also recorded greater ACE inhibition.
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Raw Peanut, Alcalase
0
2000
4000
6000
8000
10000
12000
14000
Unhydrolyzed PeptideFraction
Specific ACE Inhibition
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Raw Peanut, Pepsin-Pancreatin
0
5000
10000
15000
20000
25000
30000
35000
40000
Unhydrolyzed PeptideFraction
Specific ACE Inhibition
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RESULTS Con’t
• Preliminary studies show that hydrolysates from Alcalase have more bactericidal effects on E.coli O157:H7 and L. monocytogenes
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Antimicrobial Activity
• Without Peptides
With Peptides
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E at = M ore peanut produ C ts2
Eat = More peanut produ Cts2
E at = M ore peanut produ C ts2
E = MC2