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Cloning, Sequence Analysis, and Expression in Escherichia coli of Gene Encoding N-Benzyl-3-pyrrolidinol Dehydrogenase from Geotrichum capitatum

Paper ID Volume ID Publish Year Pages File Format Full-Text
22459 43288 2007 6 PDF Available
Title
Cloning, Sequence Analysis, and Expression in Escherichia coli of Gene Encoding N-Benzyl-3-pyrrolidinol Dehydrogenase from Geotrichum capitatum
Abstract

The gene encoding N-benzyl-3-pyrrolidinol dehydrogenase (DDBJ/EMBL/GenBank accession no. AB294179), a useful biocatalyst for producing (S)-N-benzyl-3-pyrrolidinol, was cloned from the genomic DNA of Geotrichum capitatum JCM 3908. The gene contained an open reading frame consisting of 1023 nucleotides corresponding to 340 amino acid residues. The subunit molecular weight was calculated to be 39,000. The predicted amino acid sequence did not have significant similarity to those of N-benzyl-3-pyrrolidinone reductases reported previously. From 30 mM N-benzyl-3-pyrrolidinone, (S)-N-benzyl-3-pyrrolidinol was obtained with a yield >99.9% and an enantiomeric excess >99.9% in 1-h and 2-h reactions without NADH addition by the resting cells of Escherichia coli HB 101 strains harboring the expression plasmids pSG-POBS and pSF-POBS that possess the glucose dehydrogenase gene and formate dehydrogenase gene as an NADH-reproducing system, respectively, besides the N-benzyl-3-pyrrolidinol dehydrogenase gene. N-Benzyl-3-pyrrolidinol dehydrogenase activity (0.56 U/mg) was observed in E. coli (pSG-POBS), which was 17-fold the specific activity observed in G. capitatum JCM 3908.

Keywords
N-benzyl-3-pyrrolidinol dehydrogenase; (S)-N-benzyl-3-pyrrolidinol production; N-benzyl-3-pyrrolidinone reductase; Geotrichum capitatum
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Cloning, Sequence Analysis, and Expression in Escherichia coli of Gene Encoding N-Benzyl-3-pyrrolidinol Dehydrogenase from Geotrichum capitatum
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Publisher
Database: Elsevier - ScienceDirect
Journal: Journal of Bioscience and Bioengineering - Volume 104, Issue 5, 2007, Pages 379–384
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us