title: Identification and analysis of genes from Streptomyces pristinaespiralis encoding enzymes involved in the biosynthesis of the 4-dimethylamino-L-phenylalanine precursor of pristinamycin I.
abstract: Four pap genes (papA, papB, papC, papM) were found by sequencing near to snbA, a Streptomyces pristinaespiralis gene which was previously shown to encode one of the pristinamycin I (PI) synthetases. Analysis of the homologies observed from the deduced amino acid sequences suggested that these four genes could be involved in the biosynthesis of the PI precursor 4-dimethylamino-L-phenylalanine (DMPAPA). This was first verified when disruption of papA in S. pristinaespiralis led to a PI- phenotype, which was reversed by the addition of DMPAPA into the culture medium. Further confirmation was obtained when papM was overexpressed in Escherichia coli and the corresponding protein purified to homogeneity. It catalysed the two successive N-methylation steps of 4-amino-L-phenylalanine leading to DMPAPA via 4-methylamino-L-phenylalanine. These results allowed us to assign a function to each of the four pap genes and to propose a biosynthetic pathway for DMPAPA.
authors.affiliation: Division Recherche Pharmaceutique, Centre de Recherche de Vitry-Alfortville, Vitry sur Seine, France. veronique.blanc@rp.fr
authors.forename: V
authors.initials: V
authors.lastname: Blanc
authors.forename: P
authors.initials: P
authors.lastname: Gil
authors.forename: N
authors.initials: N
authors.lastname: Bamas-Jacques
authors.forename: S
authors.initials: S
authors.lastname: Lorenzon
authors.forename: M
authors.initials: M
authors.lastname: Zagorec
authors.forename: J
authors.initials: J
authors.lastname: Schleuniger
authors.forename: D
authors.initials: D
authors.lastname: Bisch
authors.forename: F
authors.initials: F
authors.lastname: Blanche
authors.forename: L
authors.initials: L
authors.lastname: Debussche
authors.forename: J
authors.initials: J
authors.lastname: Crouzet
authors.forename: D
authors.initials: D
authors.lastname: Thibaut
mesh_terms: D003001:Cloning, Molecular; D005798:Genes, Bacterial / Q000235:genetics* / Q000502:physiology; D008969:Molecular Sequence Data; D015394:Molecular Structure; D010649:Phenylalanine / Q000031:analogs & derivatives* / Q000096:biosynthesis; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D013302:Streptomyces / Q000201:enzymology*; D014769:Virginiamycin / Q000096:biosynthesis / Q000737:chemistry
chemical_list: C105707:4-dimethylamino-phenylalanine; D014769:Virginiamycin; D010649:Phenylalanine