Optimisation and validation of an angiotensin-converting enzyme inhibition assay for the screening of bioactive peptides.
Angiotensin-converting enzyme (ACE) plays a major role in the regulation of blood pressure. A diagnostic assay to measure angiotensin-converting enzyme (ACE) activity was transformed into an enzyme inhibition assay and optimised, which led to a more sensitive and less expensive assay. By this spectrophotometric method, ACE inhibition is measured using the substrate furanacryloyl-Phe-Gly-Gly and as ACE source rabbit lung acetone extract. The optimised as well as the original ACE inhibition assay were used to verify the ACE inhibitory activity of captopril. The ACE inhibition assay was further validated by enalapril, its active derivative enalaprilat and the ACE-inhibitory peptide Ala-Leu-Pro-Met-His-Ile-Arg, corresponding to a tryptic fragment of bovine beta-lactoglobulin. Sigmoid curves could be fit adequately to the data points representing ACE inhibition in function of inhibitor concentration. IC(50) values for these compounds corresponded well with literature data. Furthermore, pea and whey protein hydrolysates obtained by digestion with trypsin showed ACE inhibitory activity in the ACE inhibition assay. Hence, this optimised assay is suitable to screen for ACE inhibitory peptides derived from food proteins with a possible antihypertensive effect in vivo.
Vermeirssen
Vanessa
V
Laboratory of Microbial Ecology and Technology, Faculty of Agricultural and Applied Biological Sciences, Ghent University Coupure Links 653, B-9000 Ghent, Belgium.
Van Camp
John
J
Verstraete
Willy
W
D000096:Acetone / Q000378:metabolism; D000806:Angiotensin-Converting Enzyme Inhibitors / Q000494:pharmacology*; D000818:Animals; D001671:Biochemistry / Q000379:methods*; D002138:Calibration; D002417:Cattle; D020128:Inhibitory Concentration 50; D007782:Lactoglobulins / Q000378:metabolism; D008168:Lung / Q000378:metabolism; D010455:Peptides / Q000737:chemistry*; D007703:Peptidyl-Dipeptidase A / Q000378:metabolism*; D011817:Rabbits; D012680:Sensitivity and Specificity
D000806:Angiotensin-Converting Enzyme Inhibitors; D007782:Lactoglobulins; D010455:Peptides; D000096:Acetone; D007703:Peptidyl-Dipeptidase A