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Enantioselective oxidation of secondary alcohols at a quinohaemoprotein alcohol dehydrogenase electrode

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Abstract

Quinohaemoprotein alcohol dehydrogenase fromComamonas testosteroni was co-immobilized with a redox polymer (a poly(vinylpyridine) complex functionalized with osmium bis(bipyridine) chloride) on an electrode. The enzyme electrode readily oxidizes primary alcohols and secondary alcohols with maximum current densities varying between 0.43 and 0.98 A m-2 depending on the substrate and the operation temperature. The affinity of the enzyme for aliphatic alcohols increases with the chain length of the substrate (i.e., 1-pentano1 [Km = 0.006 mM] is a much better substrate than ethanol [Km= 2.2 mM]). The same property is observed for secondary alcohols in the series 2-propanol (Km = 22 mM) to 2-octano1 (Km = 0.05 mM). The enzyme electrode is enantioselective in the oxidation of secondary alcohols. A strong preference is observed for the S-2-alcohols; the enantioselectivity increases with increasing chain length. The enantiomeric ratio (E) increases from 13 for (R,S)-2-butanol to approximately 80 for (R,S)-2-heptanol and (R,S)-2-octanol. This makes the enzyme electrode, potentially, a powerful tool for the preparation of a large range of alkanones and/or for the (kinetic) resolution of racemic alcohols.

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Abbreviations

BCA:

bicinchoric acid

BIS-TRIS Propane:

1, 3-bis[tris (hydroxymethyl)methylamino] propane

BSA:

bovine serum albumin

C:

concentration (mmol 1-1)

CAPS:

3-[cyclohexylamino]-l-propanesul-fonic acid

I:

current density (A m-2)

Km :

apparent Michaelis Menten con-stant (mmol 1-1)

MOPS:

3[-N-Morpholino] propane sulfonic acid

PEGDE:

polyethylene glycol diglycidylether

pI:

isoelectric point

PQQ:

pyrrolo-quinoline quinone

POs-EA:

poly(vinylpyridine) complex with osmium bis(bipyridine) chloride

QH-EDH:

quinohaemoprotein alcohol dehydrogenase

T:

temperature (%C)

TAPS:

n-tris[Hydroymethyl] methyl-3-aminopropanesulfonic acid

U:

Unit of enzyme activity (mol min-1)

Vmax :

maximum velocity (U mg-1)

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Authors and Affiliations

  1. Gist-brocades BV, P.O. Box 1, 2600, MA Delft, The Netherlands

    Wim A. C. Somers

  2. TNO Nutrition and Food Research Institute, Division of Biochemistry and Gene Technology, P.O. Box 360, 3700, AJ Zeist, The Netherlands

    Edwin C. A. Stigter, Wim van Hartingsveldt & Jan Pieter van der Lugt

Authors
  1. Wim A. C. Somers
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  2. Edwin C. A. Stigter
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  3. Wim van Hartingsveldt
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  4. Jan Pieter van der Lugt
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Correspondence to Wim van Hartingsveldt.

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Somers, W.A.C., Stigter, E.C.A., van Hartingsveldt, W. et al. Enantioselective oxidation of secondary alcohols at a quinohaemoprotein alcohol dehydrogenase electrode. Appl Biochem Biotechnol 75, 151–162 (1998). https://doi.org/10.1007/BF02787770

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  • DOI: https://doi.org/10.1007/BF02787770

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