The pyrimidine biosynthetic pathway and its regulation in Pseudomonas jessenii

Antonie Van Leeuwenhoek. 2019 Mar;112(3):461-469. doi: 10.1007/s10482-018-1168-8. Epub 2018 Sep 24.

Abstract

The control of the pyrimidine biosynthetic pathway by pyrimidine bases was examined in Pseudomonas jessenii ATCC 700870. The pyrimidine biosynthetic enzymes aspartate transcarbamoylase, dihydroorotase, dihydroorotate dehydrogenase, orotate phosphoribosyltransferase, and orotidine 5'-monophosphate (OMP) decarboxylase activities were found to be higher in the succinate-grown ATCC 700870 cells than the glucose-grown cells. All the enzyme activities were depressed in uracil-supplemented ATCC 700870 glucose-grown cells relative to the unsupplemented cells which was indicative of possible repression of enzyme synthesis by uracil. In the succinate-grown, ATCC 700870 cells, transcarbamoylase, dihydroorotase and dehydrogenase activities were decreased by uracil and orotate supplementation while decarboxylase activity was decreased following uracil addition. A pyrimidine auxotroph was isolated by conventional chemical mutagenesis and resistance to 5-fluoroorotic acid whose pyrimidine requirement was met by uracil or cytosine. The mutant strain was deficient for orotate phosphoribosyltransferase activity. Pyrimidine limitation of the mutant strain cells for 1 or 2 h caused about a two-fold increase in aspartate transcarbamoylase or dihydroorotase activity independent of carbon source relative to excess uracil growth conditions. At the level of enzyme activity, aspartate transcarbamoylase activity in P. jessenii ATCC 700870 was inhibited strongly by pyrophosphate, ATP, UTP, GTP and UMP under saturating substrate concentrations.

Keywords: Aspartate transcarbamoylase; Biosynthesis; Pseudomonas jessenii; Pyrimidines; Regulation.

MeSH terms

  • Biosynthetic Pathways / genetics*
  • Culture Media / chemistry
  • Enzymes / genetics
  • Enzymes / metabolism
  • Gene Expression Regulation, Bacterial*
  • Gene Expression Regulation, Enzymologic*
  • Mutagenesis
  • Mutation
  • Pseudomonas / genetics*
  • Pseudomonas / growth & development
  • Pseudomonas / metabolism*
  • Pyrimidines / biosynthesis*

Substances

  • Culture Media
  • Enzymes
  • Pyrimidines