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New application of firefly luciferase − it can catalyze the enantioselective thioester formation of 2‐arylpropanoic acid
Author(s) -
Kato DaiIchiro,
Teruya Keisuke,
Yoshida Hiromitsu,
Takeo Masahiro,
Negoro Seiji,
Ohta Hiromichi
Publication year - 2007
Publication title -
the febs journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.981
H-Index - 204
eISSN - 1742-4658
pISSN - 1742-464X
DOI - 10.1111/j.1742-4658.2007.05921.x
Subject(s) - thioester , luciferase , luciferases , enantioselective synthesis , chemistry , stereochemistry , enantiomer , enantiomeric excess , biochemistry , enzyme , catalysis , transfection , gene
We introduce a new application of firefly luciferase (EC 1.13.12.7). The firefly luciferases belong to a large superfamily that includes rat liver long‐chain acyl‐CoA synthetase (LACS1). LACS1 is the enzyme that is involved in the deracemization process of 2‐arylpropanoic acid and catalyzes the enantioselective thioester formation of R ‐acids. Based on the similarity of the reaction mechanisms and the sequences between firefly luciferase and LACS1, we predicted that firefly luciferase also has thioesterification activity toward 2‐arylpropanoic acid. From an investigation using three kinds of luciferases from North American firefly and Japanese fireflies, we have confirmed that these luciferases exhibit an enantioselective thioester formation activity and the R ‐form is transformed to a thioester in preference to the S ‐form in the presence of ATP, Mg 2+ , and CoASH. The enantiomeric excesses of unreacted recovered acid and thioester were determined by chiral phase HPLC analysis and the resulting 2‐arylpropanoyl‐CoAs were identified by high resolution mass spectroscopy. The K m and k cat values of thermostable luciferase from Luciola lateralis (LUC‐H) toward ketoprofen were determined as 0.22 m m and 0.11 s −1 , respectively. The affinity of ketoprofen was almost the same of d ‐luciferin. In addition, the calculated E ‐value toward ketoprofen was approximately 20. These results suggest that LUC‐H could catalyze the kinetic resolution of 2‐arylpropanoic acid efficiently and would be a new option for the preparation of optically active 2‐substituted carboxylic acids.