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J. Bacteriol., Sep 1995, 5206-5214, Vol 177, No. 18
V Blanc, D Lagneaux, P Didier, P Gil, P Lacroix and J Crouzet
In Streptomyces pristinaespiralis, two enzymes are necessary for conversion
of pristinamycin IIB (PIIB) to pristinamycin IIA (PIIA), the major
component of pristinamycin (D. Thibaut, N. Ratet, D. Bisch, D. Faucher, L.
Debussche, and F. Blanche, J. Bacteriol. 177:5199-5205, 1995); these
enzymes are PIIA synthase, a heterodimer composed of the SnaA and SnaB
proteins, which catalyzes the oxidation of PIIB to PIIA, and the
NADH:riboflavin 5'-phosphate oxidoreductase (hereafter called FMN
reductase), the SnaC protein, which provides the reduced form of flavin
mononucleotide for the reaction. By using oligonucleotide probes designed
from limited peptide sequence information of the purified proteins, the
corresponding genes were cloned from a genomic library of S.
pristinaespiralis. SnaA and SnaB showed no significant similarity with
proteins from databases, but SnaA and SnaB had similar protein domains.
Disruption of the snaA gene in S. pristinaespiralis led to accumulation of
PIIB. Complementation of a S. pristinaespiralis PIIA- PIIB+ mutant with the
snaA and snaB genes, cloned in a low-copy-number plasmid, partially
restored production of PIIA. The deduced amino acid sequence of the snaC
gene showed no similarity to the sequences of other FMN reductases but was
39% identical with the product of the actVB gene of the actinorhodin
cluster of Streptomyces coelicolor A(3)2, likely to be involved in the
dimerization step of actinorhodin biosynthesis. Furthermore, an S.
coelicolor A(3)2 mutant blocked in this step was successfully complemented
by the snaC gene, restoring the production of actinorhodin.
Copyright © 1995, American Society for Microbiology
Cloning and analysis of structural genes from Streptomyces pristinaespiralis encoding enzymes involved in the conversion of pristinamycin IIB to pristinamycin IIA (PIIA): PIIA synthase and NADH:riboflavin 5'-phosphate oxidoreductase
Unite de Biologie Moleculaire, Centre de Recherche de Vitry- Alfortville, Rhone-Poulenc Rorer S.A., Vitry-sur-Seine, France.
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