Characterization of an acidic cold-adapted cutinase from Thielavia terrestris and its application in flavor ester synthesis

Food Chem. 2015 Dec 1:188:439-45. doi: 10.1016/j.foodchem.2015.05.026. Epub 2015 May 6.

Abstract

An acidic cutinase (TtcutB) from Thielavia terrestris CAU709 was purified to apparent homogeneity with 983 Um g(-1) specific activity. The molecular mass of the enzyme was estimated to be 27.3 and 27.9 kDa by SDS-PAGE and gel filtration, respectively. A peptide sequence homology search revealed no homologous cutinases from T. terrestris, except for one putative cutinase gene (XP003656017.1), indicating that TtcutB is a novel enzyme. TtcutB exhibited an acidic pH optimum of 4.0, and stability at pH 2.5-10.5. Optimal activity was at 55 °C, it was stable up to 65 °C, and retained over 30% activity at 0 °C. Km values toward p-nitrophenyl (pNP) acetate, pNP-butyrate and pNP-caproate were 8.3, 1.1 and 0.88 mM, respectively. The cutinase exhibited strong synthetic activity on flavor ester butyl butyrate under non-aqueous environment, and the highest esterification efficiency of 95% was observed under the optimized reaction conditions. The enzyme's unique biochemical properties suggest great potential in flavor esters-producing industries.

Keywords: Characterization; Cold adaption; Cutinase; Flavor ester synthesis; Thielavia terrestris.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Butyrates / chemistry
  • Carboxylic Ester Hydrolases / metabolism*
  • Cold Temperature*
  • Electrophoresis, Polyacrylamide Gel
  • Esterification
  • Esters / chemistry*
  • Flavoring Agents / chemistry
  • Hydrogen-Ion Concentration
  • Molecular Weight
  • Sordariales / enzymology*
  • Substrate Specificity

Substances

  • Butyrates
  • Esters
  • Flavoring Agents
  • n-butyl n-butyrate
  • 4-nitrophenyl butyrate
  • Carboxylic Ester Hydrolases
  • cutinase