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Journal of Bacteriology, August 1999, p. 5060-5067, Vol. 181, No. 16
0021-9193/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Functional and Transcriptional Analyses of a Fengycin Synthetase Gene, fenC, from Bacillus subtilis

Tsuey-Pin Lin,1,2 Chyi-Liang Chen,2 Li-Kwan Chang,2 Johannes Scheng-Ming Tschen,3 and Shih-Tung Liu2,*

Graduate Institute of Microbiology and Immunology, National Yang-Ming University, Shih-Pai, Taipei 112,1 Molecular Genetics Laboratory, Department of Microbiology and Immunology, Chang-Gung University, Kwei-Shan, Taoyuan, 333,2 and Graduate Institute of Botany, National Chung-Hsing University, Taichung, 402,3 Taiwan

Received 25 January 1999/Accepted 21 May 1999

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.

* Corresponding author. Mailing address: Molecular Genetics Laboratory, Department of Microbiology and Immunology, Chang-Gung University, Kwei-Shan, Taoyuan, 333, Taiwan. Phone: 886-3-328-0292. Fax: 886-3-328-0292. E-mail: cgliu{at}mail.cgu.edu.tw.

Journal of Bacteriology, August 1999, p. 5060-5067, Vol. 181, No. 16
0021-9193/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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