Purification and properties of enantioselective ester hydrolase from a strain of Trichosporon species (DSMZ 11829)
IR@IIIM: CSIR-Indian Institute of Integrative Medicine, Jammu
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| Title |
Purification and properties of enantioselective ester hydrolase from a strain of Trichosporon species (DSMZ 11829)
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| Creator |
Vakhlu, J.
Johri, S. Verma, V. Koul, S. Parshad, R. Taneja, S.C. Qazi, G.N. |
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| Subject |
Biological Sciences
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| 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.
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| Date |
2005
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| Type |
Article
PeerReviewed |
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| Format |
application/pdf
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| 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 |
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| Relation |
http://dx.doi.org/10.1016/j.enzmictec.2005.01.037
http://iiim.csircentral.net/322/ |
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