Engineering stereoselectivity of ThDP-dependent enzymes

FEBS J. 2013 Dec;280(24):6374-94. doi: 10.1111/febs.12496. Epub 2013 Sep 13.

Abstract

Thiamine diphosphate-dependent enzymes are broadly distributed in all organisms, and they catalyse a broad range of C-C bond forming and breaking reactions. Enzymes belonging to the structural families of decarboxylases and transketolases have been particularly well investigated concerning their substrate range, mechanism of stereoselective carboligation and carbolyase reaction. Both structurally different enzyme families differ also in stereoselectivity: enzymes from the decarboxylase family are predominantly R-selective, whereas those from the transketolase family are S-selective. In recent years a key focus of our studies has been on stereoselective benzoin condensation-like 1,2-additions. Meanwhile, several S-selective variants of pyruvate decarboxylase, benzoylformate decarboxylase and 2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate (SEPHCHC) synthase as well as R-selective transketolase variants were created that allow access to a broad range of enantiocomplementary α-hydroxyketones and α,α'-dihydroxyketones. This review covers recent studies and the mechanistic understanding of stereoselective C-C bond forming thiamine diphosphate-dependent enzymes, which has been guided by structure-function analyses based on mutagenesis studies and from influences of different substrates and organic co-solvents on stereoselectivity.

Keywords: 2-hydroxypropiophenone; acetoin; acyloins; asymmetric synthesis; benzoin; catalysis; phenylacetylcarbinol; protein engineering; α,α′-dihydroxyketone; α-hydroxyketone.

Publication types

  • Research Support, Non-U.S. Gov't
  • Review

MeSH terms

  • Animals
  • Carboxy-Lyases / metabolism*
  • Catalysis
  • Humans
  • Stereoisomerism
  • Thiamine Pyrophosphate / chemistry*
  • Thiamine Pyrophosphate / metabolism*
  • Transketolase / metabolism*

Substances

  • Transketolase
  • Carboxy-Lyases
  • Thiamine Pyrophosphate