In vitro generation and stability of the lactokinin beta-lactoglobulin fragment (142-148).
The objectives of this study were to investigate the generation of beta-lactoglobulin fragment (142-148) (beta-LG f(142-148) during the hydrolysis of whey proteins, and the in vitro stability of this fragment upon incubation with gastrointestinal and serum proteinases and peptidases. An enzyme immunoassay (EIA) protocol was developed for the quantification of beta-LG f(142-148) in whey protein hydrolysates and in human blood serum. The minimum detection limit was 3 ng/mL. The level of the peptide in whey protein hydrolysates was influenced by the degree of hydrolysis (DH). As expected, highest levels of this peptide were found in hydrolysates generated with trypsin. Sequential incubation of hydrolysates at different DH values with pepsin and Corolase PP, to simulate gastrointestinal digestion, generally resulted in the degradation of beta-LG f(142-148) as determined by EIA. Reversed-phase HPLC and angiotensin-I-converting enzyme (ACE) activity assays demonstrated that synthetic beta-LG f(142-148) was rapidly degraded upon incubation with human serum. Furthermore, beta-LG f(142-148) could not be detected by EIA in the sera of 2 human volunteers following its oral ingestion or in sera from these volunteers subsequently spiked with beta-LG f(142-148). These in vitro results indicate that beta-LG f(142-148) is probably not sufficiently stable to gastrointestinal and serum proteinases and peptidases to act as an hypotensive agent in humans following oral ingestion. The in vitro methodology described herein has general application in evaluating the hypotensive potential of food protein-derived ACE inhibitory peptides.
Walsh
D J
DJ
Department of Life Sciences, University of Limerick, Ireland.
Bernard
H
H
Murray
B A
BA
MacDonald
J
J
Pentzien
A-K
AK
Wright
G A
GA
Wal
J-M
JM
Struthers
A D
AD
Meisel
H
H
Fitzgerald
R J
RJ
D000806:Angiotensin-Converting Enzyme Inhibitors / Q000494:pharmacology*; D000818:Animals; D000959:Antihypertensive Agents / Q000494:pharmacology; D002417:Cattle; D002851:Chromatography, High Pressure Liquid / Q000662:veterinary; D004063:Digestion; D004064:Digestive System / Q000201:enzymology / Q000378:metabolism*; D006801:Humans; D006868:Hydrolysis; D007124:Immunoenzyme Techniques / Q000662:veterinary; D066298:In Vitro Techniques; D007782:Lactoglobulins / Q000737:chemistry* / Q000494:pharmacology; D008894:Milk Proteins / Q000737:chemistry* / Q000378:metabolism* / Q000494:pharmacology; D010446:Peptide Fragments; D010447:Peptide Hydrolases / Q000378:metabolism; D000067816:Whey Proteins
D000806:Angiotensin-Converting Enzyme Inhibitors; D000959:Antihypertensive Agents; D007782:Lactoglobulins; D008894:Milk Proteins; D010446:Peptide Fragments; D000067816:Whey Proteins; C121294:lactokinins; D010447:Peptide Hydrolases