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Food Engineering Progress
Korean Society for Food Engineering

Food Eng. Prog. 2012; 16(3):193-198

pISSN: 1226-4768, eISSN: 2288-1247

DOI: https://doi.org/10.13050/foodengprog.2012.16.3.193

Article

Application of Convenient Chromogen-based Assay to Measurement of Protease Activity

Nam-soo Kim1,*, Jin-Soo Maeng1, Yong-Jin Cho1, Chul-Jin Kim1, Chong-Tai Kim1
1Functional Materials Research Group, Korea Food Research Institute
*Corresponding author: Nam-soo Kim, Functional Materials Research Group, Korea Food Research Institute, Baekhyun 516, Bundang, Songnam 463-746, Korea. Tel: 82-31-780-9131; Fax: 82-31-709-9876, E-mail: k9130sen@hanmail.net

ⓒ Copyright 2012 Korean Society for Food Engineering. This is an Open-Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/4.0/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Received: May 15, 2012; Revised: Jul 23, 2012; Accepted: Jul 23, 2012

Published Online: Aug 31, 2012

Abatract

This study was carried out to apply an optimized convenient assay, exploiting azo dye-bound chromogenic substrates, to measurement of protease activity. When determined for responses at varying concentrations of two substrates, azocasein and azoalbumin, using 0.5 and 5.0 mg/mL each of bovine pancreas trypsin, 3% azocasein was found to be the most appropriate substrate solution to measure protease activity. Compared with a conventional casein-Folin phenol assay, the chromogen-based protease assay exploiting 3% azocasein showed better precision to have a coefficient of variability in seven repetitive measurements less than 1.11%. When various reagent-grade and industrial proteases that showed proteinase or peptidase activities were tested by this assay at increasing enzyme concentrations, typical shape of rectangular hyperbola in activity-enzyme concentration profiles was observed. In addition, the assay of this study was suitable for activity measurement in real samples that were prepared by hydrolyzing wheat gluten and anchovy fine powder with proteases.

Keywords: application; convenient; chromogen-based assay; measurement; protease activity