Analysis of surfactin synthetase subunits in srfA mutants of Bacillus subtilis OKB105

D. Vollenbroich, N. Mehta, Peter Zuber, J. Vater, R. M. Kamp

Research output: Contribution to journalArticle

30 Citations (Scopus)

Abstract

The srfA operon of Bacillus subtilis functions in the biosynthesis of the lipopeptide antibiotic surfactin. On the basis of nucleotide sequence and genetic analysis, it is believed to encode three enzymes (E1A, E1B, and E2) that catalyze the incorporation of the surfactin substrate amino acids. Insertion, deletion, and amino acid substitution mutations of srfA were analyzed for subunit composition and activity as determined by assays of both amino acid-dependent ATP-PP(i) exchange and aminoacyl thioester formation. Insertion mutations in srfAA (encoding E1A, the subunit that incorporates Glu, Leu, and D-Leu) eliminated production and activity of all three enzymes. Deletions within srfAA and extending from srfAA to srfAB (encoding E1B, which incorporates Val, Asp, and D-Leu) abolished the activity and production of all three enzymes. Insertions between srfAA and srfAB and within srfAB eliminate the production and activity of E1B and E2. An insertion mutation in srfAC (encoding E2, which incorporates Leu) abolished the activity of E2 only. Mutations of the active serine in the putative 4'-phosphopantetheine- binding motif of the second and third domains of E1A eliminated thioester formation and severely reduced the ATP-PP(i) exchange activity of the two domains. However, the same mutation in the first domain of E1B had little effect on Val-dependent ATP-PP(i) exchange activity but abolished thioester formation. These results indicate that the coding assignments of the srfA genes are srfAA (E1A), srfAB (E1B), and srfAC (E2).

Original languageEnglish (US)
Pages (from-to)395-400
Number of pages6
JournalJournal of Bacteriology
Volume176
Issue number2
StatePublished - 1994
Externally publishedYes

Fingerprint

Bacillus subtilis
Adenosine Triphosphate
Insertional Mutagenesis
Mutation
Enzymes
Lipopeptides
Amino Acids
Amino Acid Substitution
Operon
Viperidae
Serine
Sequence Analysis
Anti-Bacterial Agents
Genes
surfactin synthetase

ASJC Scopus subject areas

  • Applied Microbiology and Biotechnology
  • Immunology

Cite this

Vollenbroich, D., Mehta, N., Zuber, P., Vater, J., & Kamp, R. M. (1994). Analysis of surfactin synthetase subunits in srfA mutants of Bacillus subtilis OKB105. Journal of Bacteriology, 176(2), 395-400.

Analysis of surfactin synthetase subunits in srfA mutants of Bacillus subtilis OKB105. / Vollenbroich, D.; Mehta, N.; Zuber, Peter; Vater, J.; Kamp, R. M.

In: Journal of Bacteriology, Vol. 176, No. 2, 1994, p. 395-400.

Research output: Contribution to journalArticle

Vollenbroich, D, Mehta, N, Zuber, P, Vater, J & Kamp, RM 1994, 'Analysis of surfactin synthetase subunits in srfA mutants of Bacillus subtilis OKB105', Journal of Bacteriology, vol. 176, no. 2, pp. 395-400.
Vollenbroich, D. ; Mehta, N. ; Zuber, Peter ; Vater, J. ; Kamp, R. M. / Analysis of surfactin synthetase subunits in srfA mutants of Bacillus subtilis OKB105. In: Journal of Bacteriology. 1994 ; Vol. 176, No. 2. pp. 395-400.
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