Stepwise enhancement of catalytic performance of haloalkane dehalogenase LinB towards β-hexachlorocyclohexane

Ryota Moriuchi; Hiroki Tanaka; Yuki Nikawadori; Mayuko Ishitsuka; Michihiro Ito; Yoshiyuki Ohtsubo; Masataka Tsuda; Jiri Damborsky; Zbynek Prokop; Yuji Nagata

doi:10.1186/s13568-014-0072-5

Stepwise enhancement of catalytic performance of haloalkane dehalogenase LinB towards β-hexachlorocyclohexane

Ryota Moriuchi, Hiroki Tanaka, Yuki Nikawadori, Mayuko Ishitsuka, Michihiro Ito, Yoshiyuki Ohtsubo, Masataka Tsuda, Jiri Damborsky, Zbynek Prokop, Yuji Nagata

Research output: Contribution to journal › Article › peer-review

8 Citations (Scopus)

Abstract

Two haloalkane dehalogenases, LinB_UT and LinB_MI, each with 296 amino acid residues, exhibit only seven amino acid residue differences between them, but LinB_MI’s catalytic performance towards β-hexachlorocyclohexane (β-HCH) is considerably higher than LinB_UT’s. To elucidate the molecular basis governing this difference, intermediate mutants between LinB_UT and LinB_MI were constructed and kinetically characterized. The activities of LinB_UT-based mutants gradually increased by cumulative mutations into LinB_UT, and the effects of the individual amino acid substitutions depended on combination with other mutations. These results indicated that LinB_UT’s β-HCH degradation activity can be enhanced in a stepwise manner by the accumulation of point mutations.

Original language	English
Article number	72
Pages (from-to)	1-10
Number of pages	10
Journal	AMB Express
Volume	4
Issue number	1
DOIs	https://doi.org/10.1186/s13568-014-0072-5
Publication status	Published - 2014 Dec 1

Keywords

Biodegradation
Haloalkane dehalogenase
Protein evolution
Xenobiotics
β-Hexachlorocyclohexane

ASJC Scopus subject areas

Biophysics
Applied Microbiology and Biotechnology

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10.1186/s13568-014-0072-5

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Stepwise enhancement of catalytic performance of haloalkane dehalogenase LinB towards β-hexachlorocyclohexane. / Moriuchi, Ryota; Tanaka, Hiroki; Nikawadori, Yuki; Ishitsuka, Mayuko; Ito, Michihiro; Ohtsubo, Yoshiyuki; Tsuda, Masataka; Damborsky, Jiri; Prokop, Zbynek; Nagata, Yuji.

In: AMB Express, Vol. 4, No. 1, 72, 01.12.2014, p. 1-10.

Research output: Contribution to journal › Article › peer-review

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abstract = "Two haloalkane dehalogenases, LinBUT and LinBMI, each with 296 amino acid residues, exhibit only seven amino acid residue differences between them, but LinBMI{\textquoteright}s catalytic performance towards β-hexachlorocyclohexane (β-HCH) is considerably higher than LinBUT{\textquoteright}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{\textquoteright}s β-HCH degradation activity can be enhanced in a stepwise manner by the accumulation of point mutations.",

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author = "Ryota Moriuchi and Hiroki Tanaka and Yuki Nikawadori and Mayuko Ishitsuka and Michihiro Ito and Yoshiyuki Ohtsubo and Masataka Tsuda and Jiri Damborsky and Zbynek Prokop and Yuji Nagata",

note = "Funding Information: This work was supported by Grant-in-Aids for Scientific Research from Ministry of Education, Culture, Sports, Science, and Technology, and Ministry of Agriculture, Forestry, and Fisheries of Japan, the Grant Agency of the Czech Republic (P503/12/0572) and the Czech Ministry of Education (LO1214). Publisher Copyright: {\textcopyright} 2014, Moriuchi et al.; licensee Springer.",

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AU - Ishitsuka, Mayuko

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AU - Ohtsubo, Yoshiyuki

AU - Tsuda, Masataka

AU - Damborsky, Jiri

AU - Prokop, Zbynek

AU - Nagata, Yuji

N1 - Funding Information: This work was supported by Grant-in-Aids for Scientific Research from Ministry of Education, Culture, Sports, Science, and Technology, and Ministry of Agriculture, Forestry, and Fisheries of Japan, the Grant Agency of the Czech Republic (P503/12/0572) and the Czech Ministry of Education (LO1214). Publisher Copyright: © 2014, Moriuchi et al.; licensee Springer.

PY - 2014/12/1

Y1 - 2014/12/1

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AB - 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.

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KW - Haloalkane dehalogenase

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KW - Xenobiotics

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Stepwise enhancement of catalytic performance of haloalkane dehalogenase LinB towards β-hexachlorocyclohexane

Abstract

Keywords

ASJC Scopus subject areas

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