Kinetic and thermodynamic investigation of enzymatic L-ascorbyl acetate synthesis

J Biotechnol. 2013 Dec;168(4):416-20. doi: 10.1016/j.jbiotec.2013.10.033. Epub 2013 Nov 7.

Abstract

Kinetics and thermodynamics of lipase-catalyzed esterification of l-ascorbic acid in acetone were investigated by using vinyl acetate as acyl donor. The results showed that l-ascorbic acid could generate inhibition effect on lipase activity. A suitable model, Ping-Pong Bi-Bi mechanism having substrate inhibition, was thus introduced to describe the enzymatic kinetics. Furthermore, the kinetic and thermodynamic parameters were calculated from a series of experimental data according to the kinetic model. The inhibition constant of L-ascorbic acid was also obtained, which seemed to imply that enhancing reaction temperature could depress the substrate inhibition. Besides, the activation energy values of the first-step and the second-step reaction were estimated to be 37.31 and 4.94 kJ/mol, respectively, demonstrating that the first-step reaction was the rate-limiting reaction and could be easily improved by enhancing temperature.

Keywords: Activation energy; Kinetic model; Ping-Pong Bi-Bi mechanism; Substrate inhibition; l-Ascorbic acid.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Ascorbic Acid / biosynthesis*
  • Ascorbic Acid / chemistry*
  • Ascorbic Acid / metabolism
  • Catalysis
  • Enzymes, Immobilized
  • Esterification
  • Eurotiales / enzymology*
  • Kinetics
  • Lipase / antagonists & inhibitors
  • Lipase / chemistry*
  • Models, Chemical
  • Substrate Specificity
  • Thermodynamics
  • Vinyl Compounds / chemistry

Substances

  • Enzymes, Immobilized
  • Vinyl Compounds
  • Lipase
  • vinyl acetate
  • Ascorbic Acid