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HERO ID
7547727
Reference Type
Journal Article
Title
Enantioselective synthesis of ethyl-(S)-3-hydroxy-3-phenylpropanoate (S-HPPE) from ethyl-3-oxo-3-phenylpropanoate using recombinant fatty acid synthase (FAS2) from Kluyveromyces lactis KCTC 7133 in Pichia pastoris GS115
Author(s)
Cha, M; Kim, EJ; Park, J; Kim, J; Kim, BG
Year
2008
Is Peer Reviewed?
Yes
Journal
Enzyme and Microbial Technology
ISSN:
0141-0229
Publisher
ELSEVIER SCIENCE INC
Location
NEW YORK
Volume
43
Issue
7
Page Numbers
480-485
DOI
10.1016/j.enzmictec.2008.08.008
Web of Science Id
WOS:000261760300003
URL
http://
://WOS:000261760300003
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Abstract
Various yeast strains were examined for the microbial reduction of ethyl-3-oxo-3-phenylpropanoate (OPPE) to ethyl-(S)-3-hydroxy-3-phenylpropanoate(S-HPPE), which is the chiral intermediate for the synthesis of a serotonin uptake inhibitor, Fluoxetine. Kluyveromyces lactis KCTC 7133 was found as the most efficient strain in terms of high yield (83% at 50mM) and high optical purity ee > 99% of S-HPPE. Based on the protein purification, activity analysis and the genomic analysis, a fatty acid synthase (FAS) was identified as the responsible beta-ketoreductase. To increase the productivity, a recombinant Pichia pastoris GS115 over-expressing FAS2 (alpha-subunit of FAS) of K lactis KCTC7133 was constructed. In the optimized media condition, the recombinant R pastoris functionally over-expressed the FAS2. Recombinant P. pastoris showed 2.3-fold higher reductase activity compared with wild type P. pastoris. With the recombinant P pastoris. the 91% yield of S-HPPE was achieved at 50 mM OPPE maintaining the high optical purity of the product (ee > 99%). (C) 2008 Elsevier Inc. All rights reserved.
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