Issue 48, 2017

Addition of a polyhistidine tag alters the regioselectivity of carbonyl reductase S1 from Candida magnoliae

Abstract

Studying enzymatic reductions of substrates with more than a single keto group is challenging, as the carbonyl reduction can create a vast array of regio- and stereoisomers. If used as reference compounds, regio- and stereopure hydroxy ketides could facilitate the characterization of reductases with unclear regio- and stereoselectivity. We have combined nonenzymatic and enzymatic reduction and oxidation steps to obtain all four regio- and stereoisomers of tert-butyl hydroxyoxohexanoates in high optical purity (enantiomeric ratio (er) of 99 : 1 for the δ-hydroxy-β-keto isomers; er of >97 : 3 for the β-hydroxy-δ-keto isomers). Furthermore, we have prepared seven of the eight possible regioisomers and diastereomers of γ-methylated hydroxyoxohexanoates. These 11 compounds allowed unraveling the complex stereoselectivity of β,δ-diketo ester reductions catalyzed by carbonyl reductase S1 from Candida magnoliae (CMCR-S1). Our analysis shows that the regio- and stereoselectivity of CMCR-S1-catalyzed reductions is highly sensitive toward modifications at the C-terminus of CMCR-S1: in addition to the expected δ-hydroxy product, the variant with a C-terminal His-tag also led to formation of β-hydroxy by-products with high optical purity.

Graphical abstract: Addition of a polyhistidine tag alters the regioselectivity of carbonyl reductase S1 from Candida magnoliae

Supplementary files

Article information

Article type
Paper
Submitted
30 Oct 2017
Accepted
23 Nov 2017
First published
23 Nov 2017

Org. Biomol. Chem., 2017,15, 10256-10264

Addition of a polyhistidine tag alters the regioselectivity of carbonyl reductase S1 from Candida magnoliae

J. Haas, M. Häckh, V. Justus, M. Müller and S. Lüdeke, Org. Biomol. Chem., 2017, 15, 10256 DOI: 10.1039/C7OB02666H

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