Transposon mutagenesis and cloning of the genes encoding the enzymes of fengycin biosynthesis inBacillus subtilis |
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| Authors: | Chyi-Liang Chen Li-Kwan Chang Yu-Sun Chang Shih-Tung Liu and Johannes Scheng-Ming Tschen |
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| Institution: | (1) Institute of Botany, National Chung-Hsing University, 402 Taichung, Taiwan;(2) Molecular Genetics Laboratory, Department of Microbiology and Immunology, Chang-Gung College of Medicine and Technology, 333 Kwei-Shan, Taoyuan, Taiwan |
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| Abstract: | A total of 20Bacillus subtilis F29-3 mutants defective in fengycin biosynthesis was obtained by Tn917 mutagenesis. Cloning and mapping results showed that the transposon in these mutants was inserted in eleven different locations on the chromosome. We were able to use the chromosomal sequence adjacent to the transposon as a probe to screen for cosmid clones containing the fengycin biosynthesis genes. One of the clones obtained, pFC660, was 46 kb long. Eight transposon insertion sites were mapped within this plasmid. Among the eleven different mutants analyzed, four mutants had Tn917 inserted in regions which encoded peptide sequences similar to part of gramicidin S synthetase, surfactin synthetase, and tyrocidine synthetase. Our results suggest that fengycin is synthesized nonribosomally by the multienzyme thiotemplate mechanism. |
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| Keywords: | Fengycin Antifungal antibiotic Lipopeptide antibiotic Nonribosomal multienzyme thiotemplate mechanism Bacillus subtilis |
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