1887

Abstract

A unique -to- racemization of arginine by coupled arginine dehydrogenases DauA and DauB encoded by the operon has been recently reported as a prerequisite for -arginine utilization as the sole source of carbon and nitrogen through -arginine catabolic pathways in . In this study, enzymic properties of the catabolic FAD-dependent -amino acid dehydrogenase DauA and the physiological functions of the operon were further characterized with other -amino acids. These results establish DauA as a -amino acid dehydrogenase of broad substrate specificity, with -Arg and -Lys as the two most effective substrates, based on the kinetic parameters. In addition, expression of is specifically induced by exogenous -Arg and -Lys, and mutations in the operon affect utilization of these two amino acids alone. The function of DauR as a repressor in the control of the operon was demonstrated by promoter activity measurements and mobility shift assays with purified His-tagged protein . The potential effect of 2-ketoarginine (2-KA) derived from -Arg deamination by DauA as a signal molecule in induction was first revealed by mutation analysis and further supported by its effect on alleviation of DauR–DNA interactions. Through sequence analysis, putative DauR operators were identified and confirmed by mutation analysis. Induction of the operon to the maximal level was found to require the -arginine-responsive regulator ArgR, as supported by the loss of inductive effect by -Arg on expression in the mutant and binding of purified ArgR to the regulatory region . In summary, this study establishes that optimal induction of the operon requires relief of DauR repression by 2-KA and activation of ArgR by -Arg as a result of -Arg racemization by the encoded DauA and DauB.

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2010-01-01
2019-08-24
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