PMID: 8955309Dec 1, 1996Paper

Cloning and characterization of the gene encoding 1-cyclohexenylcarbonyl coenzyme A reductase from Streptomyces collinus

Journal of Bacteriology
P WangK A Reynolds

Abstract

We report the cloning of the gene encoding the 1-cyclohexenylcarbonyl coenzyme A reductase (ChcA) of Streptomyces collinus, an enzyme putatively involved in the final reduction step in the formation of the cyclohexyl moiety of ansatrienin from shikimic acid. The cloned gene, with a proposed designation of chcA, encodes an 843-bp open reading frame which predicts a primary translation product of 280 amino acids and a calculated molecular mass of 29.7 kDa. Highly significant sequence similiarity extending along almost the entire length of the protein was observed with members of the short-chain alcohol dehydrogenase superfamily. The S. collinus chcA gene was overexpressed in Escherichia coli by using a bacteriophage T7 transient expression system, and a protein with a specific ChcA activity was detected. The E. coli-produced ChcA protein was purified and shown to have similar steady-state kinetics and electrophoretic mobility on sodium dodecyl sulfate-polyacrylamide gels as the enoyl-coenzyme A reductase protein prepared from S. collinus. The enzyme demonstrated the ability to catalyze, in vitro, three of the reductive steps involved in the formation of cyclohexanecarboxylic acid. An S. collinus chcA mutant, constructed by deleti...Continue Reading

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Citations

Sep 29, 2000·The Journal of Biological Chemistry·R J HeathC O Rock
Dec 24, 2011·Natural Product Reports·Qianjin KangLinquan Bai
Apr 13, 2006·Journal of Industrial Microbiology & Biotechnology·Mohini GhatgeKevin Reynolds
Mar 11, 1998·Natural Product Reports·P M Dewick
Nov 19, 1997·Natural Product Reports·H G Floss
Jan 11, 2003·Biotechnology & Genetic Engineering Reviews·T Ashton CroppKevin A Reynolds
Jan 23, 2015·Organic Letters·Ya Nan SongRen Xiang Tan

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