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Stepwise Enhancement of Catalytic Performance of Haloalkane Dehalogenase LinB towards Beta-Hexachlorocyclohexane.
Autoři | |
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Rok publikování | 2014 |
Druh | Článek v odborném periodiku |
Časopis / Zdroj | Applied Microbiology and Biotechnology Express |
Fakulta / Pracoviště MU | |
Citace | |
www | Full Text |
Doi | http://dx.doi.org/10.1186/s13568-014-0072-5 |
Obor | Biochemie |
Klíčová slova | beta -Hexachlorocyclohexane; Xenobiotics; Biodegradation; Haloalkane dehalogenase; Protein evolution |
Popis | 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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