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Updated: Jun 24, 2026
Isolation, Propagation, and Identification of Bacterial Species with Hydrocarbon Metabolizing Properties from Aquatic Habitats
Published on: December 7, 2021
Identification and cloning of a gene encoding dichloromethane dehalogenase from a methylotrophic bacterium, Bacillus
Shijin Wu1, Huaxing Zhang, Xiang Yu
1College of Biological and Environmental Engineering, Zhejiang University of Technology, 310032 Hangzhou, People's Republic of China. Wujan28@zjut.edu.cn
Abstract:
The gene dehalA encoding a novel dichloromethane dehalogenases (DehalA), has been cloned from Bacillus circulans WZ-12 CCTCC M 207006. The open reading frame of dehalA, spanning 864 bp, encoded a 288-amino acid protein that showed 85.76% identity to the dichloromethane dehalogenases of Hyphomicrobium sp. GJ21 with several commonly conserved sequences. These sequences could not be found in putative dichloromethane (DCM) dehalogenases reported from other bacteria and fungi. DehalA was expressed in Escherichia coli BL21 (DE3) from a pET28b(+) expression system and purified. The subunit molecular mass of the recombinant DehalA as estimated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis was approximately 33 kDa. Subsequent enzymatic characterization revealed that DehalA was most active in a acidic pH range at 30 degrees , which was quite different from that observed from a facultative bacterium dichloromethane dehalogenases of Methylophilus sp. strain DM11. The Michaelis-Menten constant of DCM dehalogenase was markedly lower than that of standard DCM dehalogenases.
