Two proteins with ornithine acetyltransferase activity show different functions in Streptomyces clavuligerus: Oat2 modulates clavulanic acid biosynthesis in response to arginine.
The oat2 gene, located in the clavulanic acid gene cluster in Streptomyces clavuligerus, is similar to argJ, which encodes N-acetylornithine:glutamic acid acetyltransferase activity. Purified proteins obtained by expression in Escherichia coli of the argJ and oat2 genes of S. clavuligerus posses N-acetyltransferase activity. The kinetics and substrate specificities of both proteins are very similar. Deletion of the oat2 gene did not affect the total N-acetylornithine transferase activity and slightly reduced the formation of clavulanic acid under standard culture conditions. However, the oat2 mutant produced more clavulanic acid than the parental strain in cultures supplemented with high levels (above 1 mM) of arginine. The purified S. clavuligerus ArgR protein bound the arginine box in the oat2 promoter, and the expression of oat2 was higher in mutants with a disruption in argR (arginine-deregulated), confirming that the Arg boxes of oat2 are functional in vivo. Our results suggest that the Oat2 protein or one of its reaction products has a regulatory role that modulates clavulanic acid biosynthesis in response to high arginine concentrations.
de la Fuente
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A
Area de Microbiología, Facultad de Ciencias Biológicas y Ambientales, Universidad de León, León, Spain.
Martín
J F
JF
Rodríguez-García
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A
Liras
P
P
D000123:Acetyltransferases / Q000502:physiology*; D001120:Arginine / Q000494:pharmacology*; D001426:Bacterial Proteins / Q000502:physiology*; D019818:Clavulanic Acid / Q000096:biosynthesis*; D007700:Kinetics; D008970:Molecular Weight; D013302:Streptomyces / Q000201:enzymology*; D013379:Substrate Specificity
D001426:Bacterial Proteins; D019818:Clavulanic Acid; D001120:Arginine; D000123:Acetyltransferases; C020646:glutamate acetyltransferase