Affiliation:
1. Department of Applied Biological Chemistry, Graduate School of Agricultural and Life Sciences, University of Tokyo, Tokyo, Japan
2. Department of Environmental Life Sciences, Graduate School of Life Sciences, Tohoku University, Sendai, Japan
Abstract
ABSTRACT
The enzymes LinB
UT
and LinB
MI
(LinB from
Sphingobium japonicum
UT26 and
Sphingobium
sp. MI1205, respectively) catalyze the hydrolytic dechlorination of β-hexachlorocyclohexane (β-HCH) and yield different products, 2,3,4,5,6-pentachlorocyclohexanol (PCHL) and 2,3,5,6-tetrachlorocyclohexane-1,4-diol (TCDL), respectively, despite their 98% identity in amino acid sequence. To reveal the structural basis of their different enzymatic properties, we performed site-directed mutagenesis and X-ray crystallographic studies of LinB
MI
and its seven point mutants. The mutation analysis revealed that the seven amino acid residues uniquely found in LinB
MI
were categorized into three groups based on the efficiency of the first-step (from β-HCH to PCHL) and second-step (from PCHL to TCDL) conversions. Crystal structure analyses of wild-type LinB
MI
and its seven point mutants indicated how each mutated residue contributed to the first- and second-step conversions by LinB
MI
. The dynamics simulation analyses of wild-type LinB
MI
and LinB
UT
revealed that the entrance of the substrate access tunnel of LinB
UT
was more flexible than that of LinB
MI
, which could lead to the different efficiencies of dehalogenation activity between these dehalogenases.
Publisher
American Society for Microbiology
Subject
Molecular Biology,Microbiology
Cited by
20 articles.
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