Cloning, sequence analysis and expression of the gene encoding a novel wide-spectrum amidase belonging to the amidase signature superfamily from Achromobacter xylosoxidans

Authors

  • Gang Cai,

    1. Laboratory of Molecular Microbiology, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, 300 Fenglin Road, Shanghai, PR China
    2. Graduate School of the Chinese Academy of Sciences, Beijing, PR China
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  • Songcheng Zhu,

    1. Laboratory of Molecular Microbiology, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, 300 Fenglin Road, Shanghai, PR China
    2. Graduate School of the Chinese Academy of Sciences, Beijing, PR China
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  • Xuejuan Wang,

    1. Department of Preventive Medicine, School of Public Health, Fudan University, Shanghai 200032, PR China
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  • Weihong Jiang

    Corresponding author
    1. Laboratory of Molecular Microbiology, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, 300 Fenglin Road, Shanghai, PR China
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  • Edited by S. Mongkolsuk

*Corresponding author. Tel.: +86 21 64728788. fax: +86 21 64042385., E-mail address: whjiang@sibs.ac.cn

Abstract

Amidases are very important enzymes for industrial biocatalysis. We scored a novel amidase by screening the Achromobacter xylosoxidans gene library with cephalosporin analogous amides. The gene coding for the enzyme, designated ana, was cloned, sequenced and overexpressed in Escherichia coli. Sequence analysis of ana showed it to be an amidase signature family member. Interestingly, we noted that almost all Ana homologous amidases are from human pathogens responsible for chronic lung infections. Knowing the genetic context of Ana and its homologous amidases, we suggest that they could be a part of transposon structure. Ana can efficiently hydrolyze a series of cephalosporin analogous amides, including amides with an aninine, p-nitro-aninine, and β-naphthylamine moiety, while cephalosporin could not serve as its substrate.

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