Functional and transcriptional analyses of a fengycin synthetase gene, fenC, from Bacillus subtilis

被引:52
作者
Lin, TP
Chen, CL
Chang, LK
Tschen, JSM
Liu, ST [1 ]
机构
[1] Chang Gung Univ, Dept Microbiol & Immunol, Mol Genet Lab, Tao Yuan 333, Taiwan
[2] Natl Yang Ming Univ, Grad Inst Microbiol & Immunol, Taipei 112, Taiwan
[3] Natl Chung Hsing Univ, Grad Inst Bot, Taichung 402, Taiwan
关键词
D O I
10.1128/JB.181.16.5060-5067.1999
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
A 37-kb DNA fragment containing five fengycin synthetase genes, including fenC, fenD, fenE, fenA, and fenB, was cloned and sequenced. Among these genes,fenC encodes a fengycin synthetase 2,560 amino acids long with an estimated molecular mass of 287 kDa. This protein contains two amino acid activation modules, FenC1 and FenC2, which activate L-glutamic acid and L-ornithine, respectively. Primer extension, using mRNA isolated from the log-phase cells, identified a transcription start site located 86 nucleotides upstream from the initiation codon of fenC, implying that a promoter is located upstream from the start site. Primer extension using total RNA isolated from stationary-phase cells also identified a transcription start site located 61 nucleotides upstream from the initiation codon of fenC. Gene fusion studies demonstrated that in nHA medium, the cells transcribe the fengycin synthetase genes at two different stages of cell growth. The promoter is active during the log phase, and the activity reaches the highest level during the late log phase. The activity decreases sharply but is maintained at a low level for approximately 24 h after cells enter the early stationary phase. The results of this investigation also suggest that the transcription of fenC is positively regulated during the late log phase. Results presented herein provide further insight into fengycin synthesis by B. subtilis F29-3.
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页码:5060 / 5067
页数:8
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