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Purification and properties of enantioselective ester hydrolase from a strain of Trichosporon species (DSMZ 11829)

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Title Purification and properties of enantioselective ester hydrolase from a strain of Trichosporon species (DSMZ 11829)
 
Creator Vakhlu, J.
Johri, S.
Verma, V.
Koul, S.
Parshad, R.
Taneja, S.C.
Qazi, G.N.
 
Subject Biological Sciences
 
Description An isolated strain of Trichosporon species (DSMZ 11829) was capable of producing a novel membrane-bound, enantioselective ester hydrolase. Its application in the kinetic resolution of racemic methyl ester of 6-methoxy-�-methyl-2-naphthaleneacetic acid (naproxen) into
(S)-(+)-naproxen (>99% ee, E∼500) and acyloxy derivatives of 1-chloro-3-(1-naphthyloxy)-2-propanol into (R)-alcohol (>99% ee, E∼550) have already been reported by us. The enzyme designated as TSL was extracted from the cells by toluenization and purified to homogeneity from cell free extract with 8.5% overall yield. The purified enzyme with a specific activity of 831 U/mg protein was achieved by column
chromatography using DEAE-Sepharose, phenyl-Sepharose and Sephadex G-100 resins. The purified enzyme exhibited hydrolytic activity without any noticeable decrease in the rate of hydrolysis or its enantioselectivty in the resolution of racemic naproxen into (S)-(+)-naproxen or
in the resolution of 1-chloro-3-(1-naphthyloxy)-2-propanol and 1-(6-methoxy-2-naphthyl)ethanol. Purified enzyme is a glycosylated monomer with a molecular weight of∼40 kD and isoelectric point (pI)∼4.1. TSLwas maximally active at 30 ◦C and pH 8.0. The enzymewas insensitive to serine reactive agent PMSF, metal chealator EDTA, non-ionic detergent Triton X-100 and reducing agent mercaptoethanol. The N-terminal amino acid sequence of TSL determined as Thr-Val-Thr-Thr-Pro-Thr-Leu-Ser-Ala-Asp-Ala-Lys-Lys-Gly- did not reveal any homology with the N-terminal amino acid sequence of any other known microbial ester hydrolases.
 
Date 2005
 
Type Article
PeerReviewed
 
Format application/pdf
 
Identifier http://iiim.csircentral.net/322/1/10.1016j.enzmictec.2005.01.037.pdf
Vakhlu, J. and Johri, S. and Verma, V. and Koul, S. and Parshad, R. and Taneja, S.C. and Qazi, G.N. (2005) Purification and properties of enantioselective ester hydrolase from a strain of Trichosporon species (DSMZ 11829). Enzyme and Microbial Technology, 37 (3). pp. 330-339. ISSN 01410229
 
Relation http://dx.doi.org/10.1016/j.enzmictec.2005.01.037
http://iiim.csircentral.net/322/