| RRC ID |
54962
|
| 著者 |
Kimura K, Tran LS, Do TH, Itoh Y.
|
| タイトル |
Expression of the pgsB encoding the poly-gamma-DL-glutamate synthetase of Bacillus subtilis (natto).
|
| ジャーナル |
Biosci Biotechnol Biochem
|
| Abstract |
An industrial strain of Bacillus subtilis (natto) was used to produce poly-gamma-DL-glutamate (gammaPGA), a polymer of DL-glutamate linked by a gamma-peptide bond. In spite of efforts to improve gammaPGA production by modifying the medium, little attention has been paid to the expression of the gammaPGA synthetase gene. In this study, we investigated the expression of the gammaPGA synthetic gene and the gammaPGA product under various conditions with the LacZ-fusion of the synthetic gene (pgsB-lacZ). The 5' upstream regulatory region of the pgsB gene was also investigated by constructing deletion mutations of lacZ-fusion. The pgsB-lacZ was clearly expressed in the early stationary phase and was abolished by degU gene disruption. The results showed that pgsB-lacZ expression was repressed in rich media, and that gammaPGA production was limited by the substrate supply rather than by the amount of synthetase. Adding D-glutamate to the medium reduced gammaPGA production and synthetic gene expression. The transcription start point was determined by primer extension, and it was found that up to -721 bp (translation start point = +1) of the 5' untranslated region (UTR) was required for optimal pgsB-lacZ fusion gene expression.
|
| 巻・号 |
73(5)
|
| ページ |
1149-55
|
| 公開日 |
2009-5-1
|
| DOI |
10.1271/bbb.80913
|
| PII |
JST.JSTAGE/bbb/80913
|
| PMID |
19420703
|
| MeSH |
5' Untranslated Regions / genetics
Bacillus subtilis / enzymology
Bacillus subtilis / genetics*
Bacillus subtilis / metabolism
Culture Media / pharmacology
Gene Dosage
Gene Expression / drug effects
Glutamate Synthase / genetics*
Glutamate Synthase / metabolism
Operon / genetics
Polyglutamic Acid / analogs & derivatives*
Polyglutamic Acid / biosynthesis
Polyglutamic Acid / metabolism
Recombinant Fusion Proteins / genetics
Recombinant Fusion Proteins / metabolism
Regulatory Sequences, Nucleic Acid / genetics
Sequence Deletion
Transcription, Genetic
|
| IF |
1.297
|
| 引用数 |
25
|
| リソース情報 |
| 原核生物(枯草菌) |
MBS679 |
