Transposon mutagenesis and cloning of the genes encoding the enzymes of fengycin biosynthesis in Bacillus subtilis

Chyi Liang Chen; Li Kwan Chang; Yu Sun Chang; Shih Tung Liu; Johannes Scheng Ming Tschen

doi:10.1007/BF02190792

Transposon mutagenesis and cloning of the genes encoding the enzymes of fengycin biosynthesis in Bacillus subtilis

Chyi Liang Chen
, Li Kwan Chang
, Yu Sun Chang
, Shih Tung Liu^*
, Johannes Scheng Ming Tschen

^*Corresponding author for this work

Research output: Contribution to journal › Journal Article › peer-review

33 Scopus citations

Abstract

A total of 20 Bacillus subtilis F29-3 mutants defective in fengycin biosynthesis was obtained by Tn 917 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 Tn 917 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.

Original language	English
Pages (from-to)	121-125
Number of pages	5
Journal	MGG Molecular & General Genetics
Volume	248
Issue number	2
DOIs	https://doi.org/10.1007/BF02190792
State	Published - 07 1995

Keywords

Antifungal antibiotic
Bacillus subtilis
Fengycin
Lipopeptide antibiotic
Nonribosomal multienzyme thiotemplate mechanism

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10.1007/BF02190792

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title = "Transposon mutagenesis and cloning of the genes encoding the enzymes of fengycin biosynthesis in Bacillus subtilis",

abstract = "A total of 20 Bacillus subtilis F29-3 mutants defective in fengycin biosynthesis was obtained by Tn 917 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 Tn 917 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.",

keywords = "Antifungal antibiotic, Bacillus subtilis, Fengycin, Lipopeptide antibiotic, Nonribosomal multienzyme thiotemplate mechanism",

author = "Chen, \{Chyi Liang\} and Chang, \{Li Kwan\} and Chang, \{Yu Sun\} and Liu, \{Shih Tung\} and Tschen, \{Johannes Scheng Ming\}",

year = "1995",

month = jul,

doi = "10.1007/BF02190792",

language = "英语",

volume = "248",

pages = "121--125",

journal = "MGG Molecular \& General Genetics",

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TY - JOUR

T1 - Transposon mutagenesis and cloning of the genes encoding the enzymes of fengycin biosynthesis in Bacillus subtilis

AU - Chen, Chyi Liang

AU - Chang, Li Kwan

AU - Chang, Yu Sun

AU - Liu, Shih Tung

AU - Tschen, Johannes Scheng Ming

PY - 1995/7

Y1 - 1995/7

N2 - A total of 20 Bacillus subtilis F29-3 mutants defective in fengycin biosynthesis was obtained by Tn 917 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 Tn 917 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.

AB - A total of 20 Bacillus subtilis F29-3 mutants defective in fengycin biosynthesis was obtained by Tn 917 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 Tn 917 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.

KW - Antifungal antibiotic

KW - Bacillus subtilis

KW - Fengycin

KW - Lipopeptide antibiotic

KW - Nonribosomal multienzyme thiotemplate mechanism

UR - https://www.scopus.com/pages/publications/0029116455

U2 - 10.1007/BF02190792

DO - 10.1007/BF02190792

M3 - 文章

C2 - 7651334

AN - SCOPUS:0029116455

SN - 0026-8925

VL - 248

SP - 121

EP - 125

JO - MGG Molecular & General Genetics

JF - MGG Molecular & General Genetics

IS - 2

ER -

Transposon mutagenesis and cloning of the genes encoding the enzymes of fengycin biosynthesis in Bacillus subtilis

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Keywords

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