Publication details

Stepwise Enhancement of Catalytic Performance of Haloalkane Dehalogenase LinB towards Beta-Hexachlorocyclohexane.

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Authors

MORIUCHI Ryota TANAKA Hiroki NIKAWADORI Yuki ISHITSUKA Mayuko ITO Michihiro OHTSUBO Yoshiyuki TSUDA Masataka DAMBORSKÝ Jiří PROKOP Zbyněk NAGATA Yuji

Year of publication 2014
Type Article in Periodical
Magazine / Source Applied Microbiology and Biotechnology Express
MU Faculty or unit

Faculty of Science

Citation
Web Full Text
Doi http://dx.doi.org/10.1186/s13568-014-0072-5
Field Biochemistry
Keywords beta -Hexachlorocyclohexane; Xenobiotics; Biodegradation; Haloalkane dehalogenase; Protein evolution
Description Two haloalkane dehalogenases, LinBUT and LinBMI, each with 296 amino acid (aa) residues, exhibit only seven amino acid residue differences between them, but LinBMI‘s catalytic performance towards beta-hexachlorocyclohexane (beta-HCH) is considerably higher than LinBUT‘s. To elucidate the molecular basis governing this difference, intermediate mutants between LinBUT and LinBMI were constructed and kinetically characterized. The activities of LinBUT-based mutants gradually increased by cumulative mutations into LinBUT, and the effects of the individual amino acid substitutions depended on combination with other mutations. These results indicated that LinBUT‘s b-HCH degradation activity can be enhanced in a stepwise manner by the accumulation of point mutations.
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