Premium
Identification and analysis of genes from Streptomyces pristinaespiralis encoding enzymes involved in the biosynthesis of the 4‐dimethylamino‐ l ‐phenylalanine precursor of pristinamycin I
Author(s) -
Blanc Véronique,
Gil Patricia,
BamasJacques Nathalie,
Lorenzon Sophie,
Zagorec Monique,
Schleuniger Josiane,
Bisch Didier,
Blanche Francis,
Debussche Laurent,
Crouzet Joe¨l,
Thibaut Denis
Publication year - 1997
Publication title -
molecular microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.857
H-Index - 247
eISSN - 1365-2958
pISSN - 0950-382X
DOI - 10.1046/j.1365-2958.1997.2031574.x
Subject(s) - biology , gene , biosynthesis , phenylalanine , streptomyces , amino acid , biochemistry , enzyme , methylation , escherichia coli , genetics , bacteria
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.