An Engineered Alcohol Oxidase for the Oxidation of Primary Alcohols

Research output: Contribution to journalArticle

  • External authors:
  • Rachel S. Heath
  • William R. Birmingham
  • Matthew P. Thompson
  • Andreas Taglieber
  • Laurent Daviet

Abstract

Structure-guided directed evolution of choline oxidase has been carried out by using the oxidation of hexan-1-ol to hexanal as the target reaction. A six-amino-acid variant was identified with a 20-fold increased kcat compared to that of the wild-type enzyme. This variant enabled the oxidation of 10 mm hexanol to hexanal in less than 24 h with 100 % conversion. Furthermore, this variant showed a marked increase in thermostability with a corresponding increase in Tm of 20 °C. Improved solvent tolerance was demonstrated with organic solvents including ethyl acetate, heptane and cyclohexane, thereby enabling improved conversions to the aldehyde by up to 30 % above conversion for the solvent-free system. Despite the evolution of choline oxidase towards hexan-1-ol, this new variant also showed increased specific activities (by up to 100-fold) for around 50 primary aliphatic, unsaturated, branched, cyclic, benzylic and halogenated alcohols.

Bibliographical metadata

Original languageEnglish
JournalCHEMBIOCHEM
Early online date19 Oct 2018
DOIs
Publication statusPublished - 10 Dec 2018