J. Biotechnol. 2011 Mar 20;152(3):87-92. doi: 10.1016/j.jbiotec.2011.01.020. Epub 2011 Feb 3.

Influence of reaction conditions on the enantioselectivity of biocatalyzed C-C bond formations under high pressure conditions.

S. Kara 1), W. S. Long, M. Berheide, S. Peper, B. Niemeyer, A. Liese

1)  Institute of Technical Biocatalysis, Hamburg University of Technology (TUHH), Denickestrasse 15, D-21073 Hamburg, Germany.


Abstract

Benzoylformate decarboxylase (BFD, EC 4.1.1.7) is a homotetrameric thiamine diphosphate (ThDP)-dependent enzyme which catalyzes the synthesis of chiral 2-hydroxyketones accepting a broad range of aldehydes as substrates. In this study the synthesis of 2-hydroxypropiophenone (2-HPP) from benzaldehyde and acetaldehyde was catalyzed by three BFD variants namely BFD F464I, BFD A460I and BFD A460I-F464I. This paper reports the effect of hydrostatic pressure up to 290 MPa when the reactions were carried out at different benzaldehyde concentrations (5-40 mM) as well as at different pH values (7.0-8.5). Acetaldehyde concentration was fixed at 400 mM in all biotransformations. Reactions performed at high benzaldehyde concentrations and at high hydrostatic pressures showed an increase in (R)-2-HPP formation catalyzed by all BFD variants. For BFD A460I-F464I we observed an increase in the ee of (R)-2-HPP up to 80%, whereas at atmospheric conditions this variant synthesizes (R)-2-HPP with an ee of only 50%. Alkaline conditions (up to pH 8.5) and high hydrostatic pressures resulted in an increase of (R)-2-HPP synthesis, especially in the case of BFD A460I and BFD F464I.

Copyright © 2011 Elsevier B.V. All rights reserved.

PMID: 21295624
DOI: 10.1016/j.jbiotec.2011.01.020
[Indexed for MEDLINE]