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Purification and characterization of carbonyl reductase from Geotrichum candidum

Paper ID Volume ID Publish Year Pages File Format Full-Text
35335 45087 2009 6 PDF Available
Title
Purification and characterization of carbonyl reductase from Geotrichum candidum
Abstract

Geotrichum candidum is well known for the reduction of prochiral ketones to chiral alcohol with high yield and excellent enantioselectivity. Carbonyl reductase from G. candidum was purified by ammonium sulphate precipitation, anion exchange and hydrophobic interaction chromatographies. Gel filtration chromatography together with SDS-PAGE revealed this protein to be a dimer of 60 kDa subunits. Maximum enzyme activity was found in acetate buffer at pH 5.4 with t1/2 of 7.13 h at 30 °C and t1/2 of 2.8 h at 65 °C. The enzyme was inhibited by p-hydroxymercuribenzoate and hydroxylamine indicating the involvement of thiol and carbonyl groups in the reduction reaction catalyzed by the enzyme. Chelating agents also reduced the enzyme activity indicating the requirement of metal ions as cofactors. The purified carbonyl reductase was found to be highly selective for ketones containing naphthyl ring, whereas aryl or hetero-aryl ketones showed very less or no activity at all.

Keywords
Geotrichum candidum; Carbonyl reductase; S (−)-1-(1′-naphthyl) ethanol; Purification; Characterization; Thermostability
First Page Preview
Purification and characterization of carbonyl reductase from Geotrichum candidum
Publisher
Database: Elsevier - ScienceDirect
Journal: Process Biochemistry - Volume 44, Issue 9, September 2009, Pages 986–991
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering