| RRC ID |
5017
|
| 著者 |
Yamada-Onodera K, Kojima K, Takase Y, Tani Y.
|
| タイトル |
Cloning, sequence analysis, and expression in Escherichia coli of gene encoding N-Benzyl-3-pyrrolidinol dehydrogenase from Geotrichum capitatum.
|
| ジャーナル |
J Biosci Bioeng
|
| 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.
|
| 巻・号 |
104(5)
|
| ページ |
379-84
|
| 公開日 |
2007-11-1
|
| DOI |
10.1263/jbb.104.379
|
| PII |
S1389-1723(07)70176-7
|
| PMID |
18086437
|
| MeSH |
Alcohol Oxidoreductases / biosynthesis*
Alcohol Oxidoreductases / chemistry*
Alcohol Oxidoreductases / genetics
Amino Acid Sequence
Catalysis
Cloning, Molecular
Escherichia coli / genetics
Fungal Proteins / biosynthesis*
Fungal Proteins / chemistry*
Fungal Proteins / genetics
Geotrichum / enzymology*
Molecular Sequence Data
Plasmids / genetics
Pyrroles / metabolism*
Sequence Analysis, Protein
|
| IF |
2.366
|
| 引用数 |
5
|
|
WOS 分野
|
FOOD SCIENCE & TECHNOLOGY
BIOTECHNOLOGY & APPLIED MICROBIOLOGY
|
| リソース情報 |
| 一般微生物 |
JCM3908 |
