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Development of a Spectrophotometric Method for Monitoring Angiotensin-Converting Enzyme in Dairy Products
 
Julijana Tomovska*, S. Presilski, N. Gjorgievski, N. Tomovska1, M. S. Qureshi2 and N. P. Bozinovska4
University St. Kliment Ohridski”, Faculty of Biotechnical Science, Bitola, Macedonia; 1University “St. Ciryl and Methodius”, Natural mathematics Faculty, Institute for Chemistry, Skopje, Macedonia; 2Faculty of Animal Husbandry and Veterinary Sciences, Agricultural University, Peshawar, Pakistan; 3METAROM, Food Flavouring & Colouring Industry, Sydney, Australia
*Corresponding author: dzulitomovska@yahoo.com

Abstract   

The angiotensin-converting enzyme (ACE) regulates the levels of blood pressure through generation of angiotensin-II from angiotensin-I. It is of great importance to have a reliable and yet simple method for a quantitative determination ACE inhibitory peptides in whey of milk products. A rapid, simple, sensitive and accurate spectrophotometric kinetic method has been developed for determination of ACE inhibitory peptides, using competitive inhibition. Samples of dairy product from the market were used for the determination of ACE inhibitory peptides in whey. Holmquist’s kinetic method was used for determining ACE inhibitory activity in blood serum and Ronca-Testoni method was used for the determination of ACE inhibitory activity in whey. Enzymatic inhibition activity was determined using 0.8 mmol/L FAPGG (N-[3-(Furyl) –Acryloyl]-L-Phenylalanyl Glycyl Glycyne) as the substrate in 50 mmol/L Tris buffer at pH 8.2 at 370C and a standard serum containing ACE. First, a solution of whey was mixed in a 1 to 10 ratio with serum (elevation) containing high ACE activity. The enzymatic activity was determined by monitoring the decrease in absorbance at 340 nm as result of hydrolysis of the substrate. The concentration of ACE inhibitory peptides was determined from a standard curve of inhibitor concentration versus percent of ACE inhibition. The study suggests that the method possesses good reproducibility and accuracy. The linear range enabled determination of high enzymatic activity of ACE and all ACE inhibitory peptides from dairy products act as competitive inhibitors.

Key words: Angiotensin-converting; Competitive inhibitors; Dairy products; Enzyme; Enzymatic activity; Spectrophotometry

 
   

ISSN 0253-8318 (Print)
ISSN 2074-7764 (Online)



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