Synthetically useful variants of industrial lipases from: Burkholderia cepacia and Pseudomonas fluorescens

Kazunori Yoshida, Masakazu Ono, Takahiro Yamamoto, Takashi Utsumi, Satoshi Koikeda, Tadashi Ema

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

Industrial enzymes lipase PS (LPS) and lipase AK (LAK), which originate from Burkholderia cepacia and Pseudomonas fluorescens, respectively, are synthetically useful biocatalysts. To strengthen their catalytic performances, we introduced two mutations into hot spots of the active sites (residues 287 and 290). The LPS-L287F/I290A double mutant showed high catalytic activity and enantioselectivity for poor substrates for which the wild-type enzyme showed very low activity. The LAK-V287F/I290A double mutant was also an excellent biocatalyst with expanded substrate scope, which was comparable to the LPS-L287F/I290A double mutant. Thermodynamic parameters were determined to address the origin of the high enantioselectivity of the double mutant. The ΔΔH term, but not the ΔΔS term, was predominant, which suggests that the enantioselectivity is driven by a differential energy associated with intermolecular interactions around Phe287 and Ala290. A remarkable solvent effect was observed, giving a bell-shaped profile between the E values and the log&P or ϵ values of solvents with the highest E value in i-Pr2O. This suggests that an organic solvent with appropriate hydrophobicity and polarity provides the double mutant with some flexibility that is essential for excellent catalytic performance.

Original languageEnglish
Pages (from-to)8713-8719
Number of pages7
JournalOrganic and Biomolecular Chemistry
Volume15
Issue number41
DOIs
Publication statusPublished - 2017

ASJC Scopus subject areas

  • Biochemistry
  • Physical and Theoretical Chemistry
  • Organic Chemistry

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