Cloning of a Novel Dehalogenase from Environmental DNA

Hisashi KIMOTO; Shin-ichiro SUYE; Hirokazu MAKISHIMA; Jun-ichirou ARAI; Sachiko YAMAGUCHI; Yutaka FUJII; Toshihito YOSHIOKA; Akira TAKETO

doi:10.1271/bbb.100027

Biochemistry & Molecular Biology Notes

Cloning of a Novel Dehalogenase from Environmental DNA

Hisashi KIMOTO, Shin-ichiro SUYE, Hirokazu MAKISHIMA, Jun-ichirou ARAI, Sachiko YAMAGUCHI, Yutaka FUJII, Toshihito YOSHIOKA, Akira TAKETO

Author information

Hisashi KIMOTO
Department of Bioscience, Fukui Prefectural University
Shin-ichiro SUYE
Department of Applied Chemistry and Biotechnology, Graduate School of Engineering, University of Fukui
Hirokazu MAKISHIMA
Department of Applied Chemistry and Biotechnology, Graduate School of Engineering, University of Fukui
Jun-ichirou ARAI
Daikin Industries, Ltd., Environmental Technology Laboratory
Sachiko YAMAGUCHI
Daikin Industries, Ltd., Environmental Technology Laboratory
Yutaka FUJII
Department of Molecular Biology and Chemistry, Faculty of Medicine, University of Fukui
Toshihito YOSHIOKA
Department of Bioscience, Fukui Prefectural University
Akira TAKETO
Department of Environmental and Biotechnological Frontier Engineering, Fukui University of Technology

Keywords: dehalogenase, metagenome, tetrachloroethene, tetrachloroethylene, DnaK

JOURNAL FREE ACCESS

2010 Volume 74 Issue 6 Pages 1290-1292

DOI https://doi.org/10.1271/bbb.100027

View "Advance Publication" version (June 7, 2010).

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Abstract

Cloning of pceA, the gene of tetrachloroethene (PCE)-reductive dehalogenase, was undertaken from environmental DNA. Two genes were amplified using PCR primer deduced from pceA. Functional expression of these genes was unsuccessful in Escherichia coli, but PceA1 synthesized in vitro was enzymatically active. In recombinant E. coli, PceA1 formed a complex with host DnaK and caused filamentous growth.

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