Analysis of a mutant amino acid-activating domain of surfactin synthetase bearing a serine-to-alanine substitution at the site of carboxylthioester formation

Dirk Vollenbroich, Britta Kluge, Cletus D'Souza, Peter Zuber, Joachim Vater

Research output: Contribution to journalArticle

21 Citations (Scopus)

Abstract

The reactive serine of the TGGHSL thioester binding motif of the first amino acid-activating domain of surfactin synthetase was replaced by alanine using site-directed mutagenesis. The multienzyme from cells of the resulting mutant lost its ability for thioester formation with l-Glu and was therefore inactive in surfactin production. The thiolation reactions catalyzed by the other amino acid-activating domains of surfactin synthetase were not affected by the mutation. The results show that l-Glu is acativated at the first domain of surfactin synthetase, and give further evidence that a serine residue is essential for substrate amino acid activation at the reaction centers of peptide synthetases.

Original languageEnglish (US)
Pages (from-to)220-224
Number of pages5
JournalFEBS Letters
Volume325
Issue number3
DOIs
StatePublished - Jul 5 1993
Externally publishedYes

Fingerprint

Bearings (structural)
Alanine
Serine
Substitution reactions
Amino Acids
Peptide Synthases
Mutagenesis
Amino Acid Motifs
Essential Amino Acids
Site-Directed Mutagenesis
Chemical activation
Mutation
Substrates
surfactin synthetase

Keywords

  • Bacillus subtilis, Site-directed mutagenesis
  • Surfactin synthetase
  • Thioester binding: Reactive serine: Multiple carrier model

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

Analysis of a mutant amino acid-activating domain of surfactin synthetase bearing a serine-to-alanine substitution at the site of carboxylthioester formation. / Vollenbroich, Dirk; Kluge, Britta; D'Souza, Cletus; Zuber, Peter; Vater, Joachim.

In: FEBS Letters, Vol. 325, No. 3, 05.07.1993, p. 220-224.

Research output: Contribution to journalArticle

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AU - Kluge, Britta

AU - D'Souza, Cletus

AU - Zuber, Peter

AU - Vater, Joachim

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