Engineering of highly selective variants of Parvibaculum lavamentivorans alcohol dehydrogenase

Chembiochem. 2014 Sep 22;15(14):2050-2. doi: 10.1002/cbic.201402216. Epub 2014 Aug 28.

Abstract

We present the development of highly selective variants of the Parvibaculum lavamentivorans alcohol dehydrogenase. Four amino acids (A158, N162, K202, L224) in the second sphere of the catalytic site were identified to determine the selectivity for 3-quinuclidone reduction significantly. The best variant (A158H/N162G/K202Q/L224W) was able to increase the ee for (R)-3-quinuclidinol production from 84.3 % (wild-type) to ≥99 % and concomitantly to enhance conversion by 43.5 %.

Keywords: biocatalysis; high-throughput screening; oxidoreductases; protein engineering; selectivity.

MeSH terms

  • Alcohol Dehydrogenase / chemistry
  • Alcohol Dehydrogenase / genetics*
  • Alcohol Dehydrogenase / metabolism
  • Alphaproteobacteria / chemistry
  • Alphaproteobacteria / enzymology*
  • Alphaproteobacteria / genetics
  • Alphaproteobacteria / metabolism
  • Catalytic Domain
  • Models, Molecular
  • Mutation
  • Oxidation-Reduction
  • Protein Engineering*
  • Quinuclidines / metabolism*

Substances

  • Quinuclidines
  • 3-quinuclidinol
  • Alcohol Dehydrogenase