A Liquid-Based Colorimetric Assay of Lysine Decarboxylase and Its Application to Enzymatic Assay.
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.
Department of Biological Engineering, College of Engineering, Konkuk University, Seoul 143-701, Republic of Korea.
Yong Hyun
YH
Kim
Department of Biological Engineering, College of Engineering, Konkuk University, Seoul 143-701, Republic of Korea.
Ganesan
G
Sathiyanarayanan
Department of Biological Engineering, College of Engineering, Konkuk University, Seoul 143-701, Republic of Korea.
Hyun Joong
HJ
Kim
Department of Biological Engineering, College of Engineering, Konkuk University, Seoul 143-701, Republic of Korea.
Shashi Kant
SK
Bhatia
Department of Biological Engineering, College of Engineering, Konkuk University, Seoul 143-701, Republic of Korea.
Hyung-Min
HM
Seo
Department of Biological Engineering, College of Engineering, Konkuk University, Seoul 143-701, Republic of Korea.
Jung-Ho
JH
Kim
Department of Biological Engineering, College of Engineering, Konkuk University, Seoul 143-701, Republic of Korea.
Hun-Seok
HS
Song
Chemical Engineering, Soongsil University, Seoul 156-743, Republic of Korea.
Yun-Gon
YG
Kim
Department of Biological and Chemical Engineering, Hongik University, Sejong 339-701, Republic of Korea.
Kyungmoon
K
Park
Department of Biological Engineering, College of Engineering, Konkuk University, Seoul 143-701, Republic of Korea.|Microbial Carbohydrate Resource Bank, Konkuk University, Seoul 143-701, Republic of Korea.
Yung-Hun
YH
Yang
D001962:Bromcresol Purple; D002103:Cadaverine / Q000378:metabolism*; D002262:Carboxy-Lyases / Q000032:analysis*; D003124:Colorimetry / Q000379:methods*; D006863:Hydrogen-Ion Concentration; D007202:Indicators and Reagents
D007202:Indicators and Reagents; D001962:Bromcresol Purple; D002262:Carboxy-Lyases; C021520:lysine decarboxylase; D002103:Cadaverine
Bromocresol purple; Cadaverine; High-throughput colorimetric assay; Lysine decarboxylase; pH indicator