Purification of the fengycin synthetase multienzyme system from Bacillus subtilis b213

The purification of the multienzyme system producing the lipodecapeptide fe ngycin in Bacillus subtilis b213 was investigated. By gel filtration of a c ell free extract of this organism three enzyme fractions were obtained from which five multifunctional components of fengycin synthetase were separate d by high resolution anion-exchange FPLC procedures. These proteins were ch aracterized by their thioester formation activities with C-14-labeled subst rate amino acids and by N-terminal sequencing. Correlation of these data wi th the DNA sequences of the pps (fen) operons in three B. subtilis strains provided detailed knowledge on the structural and functional organization o f fengycin synthetase. (C) 2000 Elsevier Science B.V. All rights reserved.