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http://dx.doi.org/10.4014/jmb.1505.05063

A Liquid-Based Colorimetric Assay of Lysine Decarboxylase and Its Application to Enzymatic Assay  

Kim, Yong Hyun (Department of Biological Engineering, College of Engineering, Konkuk University)
Sathiyanarayanan, Ganesan (Department of Biological Engineering, College of Engineering, Konkuk University)
Kim, Hyun Joong (Department of Biological Engineering, College of Engineering, Konkuk University)
Bhatia, Shashi Kant (Department of Biological Engineering, College of Engineering, Konkuk University)
Seo, Hyung-Min (Department of Biological Engineering, College of Engineering, Konkuk University)
Kim, Jung-Ho (Department of Biological Engineering, College of Engineering, Konkuk University)
Song, Hun-Seok (Department of Biological Engineering, College of Engineering, Konkuk University)
Kim, Yun-Gon (Chemical Engineering, Soongsil University)
Park, Kyungmoon (Department of Biological and Chemical Engineering, Hongik University)
Yang, Yung-Hun (Department of Biological Engineering, College of Engineering, Konkuk University)
Publication Information
Journal of Microbiology and Biotechnology / v.25, no.12, 2015 , pp. 2110-2115 More about this Journal
Abstract
A liquid-based colorimetric assay using a pH indicator was introduced for high-throughput monitoring of lysine decarboxylase activity. The assay is based on the color change of bromocresol purple, measured at 595 nm in liquid reaction mixture, due to an increase of pH by the production of cadaverine. Bromocresol purple was selected as the indicator because it has higher sensitivity than bromothymol blue and pheonol red within a broad range and shows good linearity within the applied pH. We applied this for simple determination of lysine decarboxylase reusability using 96-well plates, and optimization of conditions for enzyme overexpression with different concentrations of IPTG on lysine decarboxylase. This assay is expected to be applied for monitoring and quantifying the liquid-based enzyme reaction in biotransformation of decarboxylase in a high-throughput way.
Keywords
Cadaverine; lysine decarboxylase; pH indicator; high-throughput colorimetric assay; bromocresol purple;
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Times Cited By KSCI : 1  (Citation Analysis)
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