Engineering the substrate-binding domain of an esterase enhances its hydrolytic activity toward fatty acid esters

Young A. Lee, Eun Yeong Jeon, Sun Mee Lee, Uwe T. Bornscheuer, Jin Byung Park

Research output: Contribution to journalArticlepeer-review

10 Scopus citations

Abstract

The poor solubility and dispersibility of fatty acids in aqueous reaction media may limit the catalytic activity of fatty acid transformation enzymes. Therefore, we studied a novel method to increase the catalytic activity of an esterase by introducing a presumed substrate-binding domain. The primary structure of an esterase from Pseudomonas fluorescens WI SIK (PFEI) is similar to that of an esterase in P. fluorescens DSM 50106 (PFEII) but not Bacillus subtilis DSM 402 (BS2). However, the reaction kinetics for the formation of octylacetate and a ricinoleic acid-derived ester (3) were more similar to the kinetics in BS2. For instance, the kcat value of PFEI with 3 was similar to that of BS2, which was approximately 12-fold lower than the kcat value of PFEII. Furthermore, fusion of PFEI to the N-terminal hydrophobic domain of PFEII led to a substantial increase (an approximate 6-fold increase in the kcat value) in its hydrolytic activity of 3. These results indicate that the N-terminal domain of PFEII, which is assumed to be involved in anchoring the enzyme in the membrane, interacts with fatty acid-like substrates, resulting in an improved enzymatic activity. Therefore, we conclude that the membrane-anchoring domains can be used to increase the catalytic activity of fatty acid transformation enzymes.

Original languageEnglish
Pages (from-to)2101-2106
Number of pages6
JournalProcess Biochemistry
Volume49
Issue number12
DOIs
StatePublished - Dec 2014

Keywords

  • Enzyme catalysis
  • Enzyme engineering
  • Esterase
  • Esterification
  • Fatty acids

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