Genetic regulation, biochemical properties and physiological importance of arginase from Free

Abstract

In bacteria, -arginine is a precursor of various metabolites and can serve as a source of carbon and/or nitrogen. Arginine catabolism by arginase, which hydrolyzes arginine to -ornithine and urea, is common in nature but has not been studied in symbiotic nitrogen-fixing rhizobia. The genome of the alfalfa microsymbiont 1021 has two genes annotated as arginases, () and (). Biochemical assays with purified ArgI1 and ArgI2 (as 6His-Sumo-tagged proteins) showed that only ArgI1 had detectable arginase activity. A 1021 null mutant lacked arginase activity and grew at a drastically reduced rate with arginine as sole nitrogen source. Wild-type growth and arginase activity were restored in the mutant genetically complemented with a genomically integrated gene. In the wild-type, arginase activity and transcription were induced several fold by exogenous arginine. ArgI1 purified as a 6His-Sumo-tagged protein had its highest enzymatic activity at pH 7.5 with Ni as cofactor. The enzyme was also active with Mn and Co, both of which gave the enzyme the highest activities at a more alkaline pH. The 6His-Sumo-ArgI1 comprised three identical subunits based on the migration of the urea-dissociated protein in a native polyacrylamide gel. A Lrp-like regulator () divergently transcribed from was required for arginase induction by arginine or ornithine. This regulator was designated ArgIR. Electrophoretic mobility shift assays showed that purified ArgIR bound to the promoter in a region preceding the predicted transcriptional start. Our results indicate that ArgI1 is the sole arginase in , that it contributes substantially to arginine catabolism and that induction by arginine is dependent on ArgIR.

Funding
This study was supported by the:
  • CONACyT (Award 290118)
    • Principle Award Recipient: Alejandra Arteaga Ide
  • DGAPA-PAPIIT (Award IN210114)
    • Principle Award Recipient: Michael F. Dunn
  • DGAPA-PAPIIT (Award IN208811)
    • Principle Award Recipient: Michael F. Dunn
  • DGAPA-PAPIIT (Award IN206317)
    • Principle Award Recipient: Michael F. Dunn
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2020-03-27
2024-03-28
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