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

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 basi...

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Detalles Bibliográficos
Autores principales: Moriuchi, Ryota, Tanaka, Hiroki, Nikawadori, Yuki, Ishitsuka, Mayuko, Ito, Michihiro, Ohtsubo, Yoshiyuki, Tsuda, Masataka, Damborsky, Jiri, Prokop, Zbynek, Nagata, Yuji
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Springer 2014
Materias:
Original Article
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4230811/
https://www.ncbi.nlm.nih.gov/pubmed/25401073
http://dx.doi.org/10.1186/s13568-014-0072-5
Descripción
Sumario: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.

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