the production of bioactive peptides from peanuts -...

FOOD Product Innovation and CommercializationFOOD Product Innovation and Commercialization

The Production of Bioactive Peptides from Peanuts

FOOD Product Innovation and CommercializationFOOD Product Innovation and Commercialization

Fat

Protein

Carbohydrate

Water

Other

Composition of peanut kernel

FOOD Product Innovation and CommercializationFOOD Product Innovation and Commercialization

Protein Structure

FOOD Product Innovation and CommercializationFOOD Product Innovation and Commercialization

Peptide Hydrolysis

FOOD Product Innovation and CommercializationFOOD Product Innovation and Commercialization

FOOD Product Innovation and CommercializationFOOD Product Innovation and Commercialization

Peptides

FOOD Product Innovation and CommercializationFOOD Product Innovation and Commercialization

Amino Acid

FOOD Product Innovation and CommercializationFOOD Product Innovation and Commercialization

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

FOOD Product Innovation and CommercializationFOOD Product Innovation and Commercialization

Peptide structure

FOOD Product Innovation and CommercializationFOOD Product Innovation and Commercialization

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

FOOD Product Innovation and CommercializationFOOD Product Innovation and Commercialization

FOOD Product Innovation and CommercializationFOOD Product Innovation and Commercialization

S t r e s s

FOOD Product Innovation and CommercializationFOOD Product Innovation and Commercialization

S t r e s s

FOOD Product Innovation and CommercializationFOOD Product Innovation and Commercialization

Up Regulation of BP by ACE

Blood Pressure

Angiotensin I Angiotensin II

* Angiotensin I Converting Enzyme

ACE*

FOOD Product Innovation and CommercializationFOOD Product Innovation and Commercialization

Down Regulation of BP by ACE InhibitorsBlood Pressure

Angiotensin I Angiotensin II

* Angiotensin I Converting Enzyme

ACE*

ACE INHIBITORS

FOOD Product Innovation and CommercializationFOOD Product Innovation and Commercialization

OBJECTIVES

• To identify and isolate hypotensive peptides from proteolytic peanut digests

• To detect and evaluate bactericidal peptides from proteolytic peanut digests

FOOD Product Innovation and CommercializationFOOD Product Innovation and Commercialization

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

FOOD Product Innovation and CommercializationFOOD Product Innovation and Commercialization

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

FOOD Product Innovation and CommercializationFOOD Product Innovation and Commercialization

Isolation of BiopeptidesCrude Hydrolysate

FOOD Product Innovation and CommercializationFOOD Product Innovation and Commercialization

METHODOLOGY Con’t

• Proteolytic digests tested against – Angiotensin Converting Enzyme (ACE) for

antihypertensive activity – E.coli O157:H7 and L. monocytogenes for

antimicrobial activity

FOOD Product Innovation and CommercializationFOOD Product Innovation and Commercialization

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

FOOD Product Innovation and CommercializationFOOD Product Innovation and Commercialization

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.

FOOD Product Innovation and CommercializationFOOD Product Innovation and Commercialization

Raw Peanut, Alcalase

0

2000

4000

6000

8000

10000

12000

14000

Unhydrolyzed PeptideFraction

Specific ACE Inhibition

FOOD Product Innovation and CommercializationFOOD Product Innovation and Commercialization

Raw Peanut, Pepsin-Pancreatin

0

5000

10000

15000

20000

25000

30000

35000

40000

Unhydrolyzed PeptideFraction

Specific ACE Inhibition

FOOD Product Innovation and CommercializationFOOD Product Innovation and Commercialization

RESULTS Con’t

• Preliminary studies show that hydrolysates from Alcalase have more bactericidal effects on E.coli O157:H7 and L. monocytogenes

FOOD Product Innovation and CommercializationFOOD Product Innovation and Commercialization

Antimicrobial Activity

• Without Peptides

With Peptides

FOOD Product Innovation and CommercializationFOOD Product Innovation and Commercialization

E at = M ore peanut produ C ts2

Eat = More peanut produ Cts2

E at = M ore peanut produ C ts2

E = MC2