Immobilization of membrane-bounded (S)-mandelate dehydrogenase in sol-gel matrix for electroenzymatic synthesis

Bioelectrochemistry. 2015 Aug:104:65-70. doi: 10.1016/j.bioelechem.2015.03.004. Epub 2015 Mar 25.

Abstract

Membrane-bounded (S)-mandelate dehydrogenase has been immobilized on the surface of glassy carbon and carbon felt electrodes by encapsulation in a silica film obtained by sol-gel chemistry. Such bioelectrochemical system has been used for the first time for electroenzymatic conversion of (S)-mandelic acid to phenylglyoxylic acid. Apparent Km in this sol-gel matrix was 0.7 mM in the presence of ferrocenedimethanol, a value in the same order of magnitude as reported previously for vesicles in solution with other electron acceptors, i.e., Fe(CN)6(3-) or 2,6-dichloroindophenol. The bioelectrode shows very good operational stability for more than 6 days. This stability was definitively improved by comparison to a bioelectrode prepared by simple adsorption of the proteins on the electrode surface (fast activity decrease during the first 15 h of experiment). Optimal electroenzymatic reaction was achieved at pH9 and 40 °C. Apparent Km of the protein activity was 3 times higher in carbon felt electrode than on glassy carbon surface, possibly because of transport limitations in the porous architecture of the carbon felt. A good correlation was found between electrochemical data and chromatographic characterization of the reaction products in the bioelectrochemical reactor.

Keywords: Bioelectrochemical system; Electrosynthesis; Membrane protein; Sol–gel.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Alcohol Oxidoreductases / chemistry*
  • Alcohol Oxidoreductases / metabolism
  • Carbon / chemistry
  • Electrochemistry
  • Electrodes
  • Enzymes, Immobilized / chemistry*
  • Enzymes, Immobilized / metabolism
  • Glass / chemistry
  • Glyoxylates / chemistry
  • Mandelic Acids / chemistry
  • Membranes, Artificial*
  • Silicon Dioxide / chemistry

Substances

  • Enzymes, Immobilized
  • Glyoxylates
  • Mandelic Acids
  • Membranes, Artificial
  • phenylglyoxylic acid
  • Carbon
  • Silicon Dioxide
  • Alcohol Oxidoreductases
  • L-mandelate dehydrogenase
  • mandelic acid