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Applied and Environmental Microbiology, December 1999, p. 5198-5206, Vol. 65, No. 12
0099-2240/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Soluble Methane Monooxygenase Gene Clusters from Trichloroethylene-Degrading Methylomonas sp. Strains and Detection of Methanotrophs during In Situ Bioremediation

Toru Shigematsu,1,* Satoshi Hanada,1 Masahiro Eguchi,2 Yoichi Kamagata,1 Takahiro Kanagawa,1 and Ryuichiro Kurane1

National Institute of Bioscience and Human-Technology, Agency of Industrial Science and Technology, Tsukuba, Ibaraki 305-8566,1 and Central Research Laboratories, Organo Corporation, Saitama 335-0015,2 Japan

Received 19 April 1999/Accepted 10 September 1999

The soluble MMO (sMMO) gene clusters from group I methanotrophs were characterized. An 8.1-kb KpnI fragment from Methylomonas sp. strain KSWIII and a 7.5-kb SalI fragment from Methylomonas sp. strain KSPIII which contained the sMMO gene clusters were cloned and sequenced. The sequences of these two fragments were almost identical. The sMMO gene clusters in the fragment consisted of six open reading frames which were 52 to 79% similar to the corresponding genes of previously described sMMO gene clusters of the group II and group X methanotrophs. The phylogenetic analysis of the predicted amino acid sequences of sMMO demonstrated that the sMMOs from these strains were closer to that from M. capsulatus Bath in the group X methanotrophs than to those from Methylosinus trichosporium OB3b and Methylocystis sp. strain M in the group II methanotrophs. Based on the sequence data of sMMO genes of our strains and other methanotrophs, we designed a new PCR primer to amplify sMMO gene fragments of all the known methanotrophs harboring the mmoX gene. The primer set was successfully used for detecting methanotrophs in the groundwater of trichloroethylene-contaminated sites during in situ-biostimulation treatments.

* Corresponding author. Present address: Department of Applied Chemistry and Biochemistry, Faculty of Engineering, Kumamoto University, Kurokami 2-39-1, Kumamoto-City, Kumamoto 860-8555, Japan. Phone: 81-96-342-3668. Fax: 81-96-342-3679. E-mail: shige{at}kumamoto-u.ac.jp.

Applied and Environmental Microbiology, December 1999, p. 5198-5206, Vol. 65, No. 12
0099-2240/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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Copyright © 1999 by the American Society for Microbiology. All rights reserved.