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Talanta. 2009 Sep 15;79(4):1130-4. doi: 10.1016/j.talanta.2009.02.016. Epub 2009 Feb 20.

Flow injection analysis of angiotensin I-converting enzyme inhibitory activity with enzymatic reactors.

Author information

1
Faculty of Agriculture, Kochi University, Monobe B-200, Nankoku 783-8502, Japan.

Abstract

Assay of angiotensin I-converting enzyme (ACE) inhibitory activity always draws much attention because of diverse applications in the field of antihypertension and related pathogenesis. Recently, the use of a new synthetic substrate, 3-hydroxybutyrylglycyl-glycyl-glycine (3HB-GGG), for the assay of ACE inhibitory activity has been confirmed. To construct a rapid, economical, and automatic determination system of ACE inhibitory activity using 3HB-GGG, a flow injection analysis (FIA) system with enzymatic reactors was developed in this study. Enzyme reactors were composed of aminoacylase and 3-hydroxybutyrate dehydrogenase immobilized separately on CNBr-activated Sepharose 4B. The assay condition was optimized in terms of the conversion of 3HB-G into NADH by the enzymatic reactors when the reaction solution containing 3HB-G generated from 3HB-GGG (after the incubation with ACE) was repetitively injected into the FIA system. Under the optimized conditions, 3HB-G was converted to 3HB, and then 3HB was oxidized by NAD(+) to form NADH. The developed system successfully detected practical ACE inhibitors with a great sensitivity, high sampling frequency (10 samples h(-1)) and a durable stability of the enzymatic reactors.

PMID:
19615521
DOI:
10.1016/j.talanta.2009.02.016
[Indexed for MEDLINE]

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