Two haloalkane dehalogenases, LinBUT and LinBMI, each with 296 amino acid residues, exhibit only seven amino acid residue differences between them, but LinBMI's catalytic performance towards β-hexachlorocyclohexane (β-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 β-HCH degradation activity can be enhanced in a stepwise manner by the accumulation of point mutations.

Department of Environmental Life Sciences Graduate School of Life Sciences Tohoku University Sendai Japan

Department of Environmental Life Sciences Graduate School of Life Sciences Tohoku University Sendai Japan ; Consolidated Research Institute for Advanced Science and Medical Care Waseda University 2 2 Wakamatsu cho Shinjuku Tokyo 162 8480 Japan

Department of Environmental Life Sciences Graduate School of Life Sciences Tohoku University Sendai Japan ; The United Graduate School of Agricultural Science Gifu University 1 1 Yanagido Gifu 501 1193 Japan

Loschmidt Laboratories Department of Experimental Biology and Research Centre for Toxic Compounds in the Environment RECETOX Faculty of Science Masaryk University Kamenice 5 A13 Brno 625 00 Czech Republic

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