RRC ID 54962
Author Kimura K, Tran LS, Do TH, Itoh Y.
Title Expression of the pgsB encoding the poly-gamma-DL-glutamate synthetase of Bacillus subtilis (natto).
Journal 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.
Volume 73(5)
Pages 1149-55
Published 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
Times Cited 25
Prokaryotes B. subtilis MBS679