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Applied and Environmental Microbiology, March 2002, p. 1220-1227, Vol. 68, No. 3
0099-2240/02/$04.00+0     DOI: 10.1128/AEM.68.3.1220-1227.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Nucleotide Sequence and Genetic Structure of a Novel Carbaryl Hydrolase Gene (cehA) from Rhizobium sp. Strain AC100

Masayuki Hashimoto,1 Mitsuru Fukui,2 Kouichi Hayano,2 and Masahito Hayatsu2*

Bio-Oriented Technology Research Advancement Institution, Minatoku, Tokyo 105-0001,1 Faculty of Agriculture, Shizuoka University, Shizuoka 422-8529, Japan2

Received 27 August 2001/ Accepted 6 December 2001

Rhizobium sp. strain AC100, which is capable of degrading carbaryl (1-naphthyl-N-methylcarbamate), was isolated from soil treated with carbaryl. This bacterium hydrolyzed carbaryl to 1-naphthol and methylamine. Carbaryl hydrolase from the strain was purified to homogeneity, and its N-terminal sequence, molecular mass (82 kDa), and enzymatic properties were determined. The purified enzyme hydrolyzed 1-naphthyl acetate and 4-nitrophenyl acetate indicating that the enzyme is an esterase. We then cloned the carbaryl hydrolase gene (cehA) from the plasmid DNA of the strain and determined the nucleotide sequence of the 10-kb region containing cehA. No homologous sequences were found by a database homology search using the nucleotide and deduced amino acid sequences of the cehA gene. Six open reading frames including the cehA gene were found in the 10-kb region, and sequencing analysis shows that the cehA gene is flanked by two copies of insertion sequence-like sequence, suggesting that it makes part of a composite transposon.

* Corresponding author: Mailing address: Faculty of Agriculture, Shizuoka University, 836 Ohya, Shizuoka 422-8529, Japan. Phone and fax: 81-54-238-4875. E-mail: ahmhaya{at}agr.shizuoka.ac.jp.

Applied and Environmental Microbiology, March 2002, p. 1220-1227, Vol. 68, No. 3
0099-2240/02/$04.00+0     DOI: 10.1128/AEM.68.3.1220-1227.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.





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