Analyses of variants of the Ser/Thr dehydratase IlvA provide insight into 2-aminoacrylate metabolism in Salmonella enterica

J Biol Chem. 2018 Dec 14;293(50):19240-19249. doi: 10.1074/jbc.RA118.005626. Epub 2018 Oct 16.


RidA is a conserved and broadly distributed protein that has enamine deaminase activity. In a variety of organisms tested thus far, lack of RidA results in the accumulation of the reactive metabolite 2-aminoacrylate (2AA), an obligate intermediate in the catalytic mechanism of several pyridoxal 5'-phosphate (PLP)-dependent enzymes. This study reports the characterization of variants of the biosynthetic serine/threonine dehydratase (EC; IlvA), which is a significant generator of 2AA in the bacteria Salmonella enterica, Escherichia coli, and Pseudomonas aeruginosa and the yeast Saccharomyces cerevisiae Two previously identified mutations, ilvA3210 and ilvA3211, suppressed the phenotypic growth consequences of 2AA accumulation in S. enterica Characterization of the respective protein variants suggested that they affect 2AA metabolism in vivo by two different catalytic mechanisms, both leading to an overall reduction in serine dehydratase activity. To emphasize the physiological relevance of the in vitro enzyme characterization, we sought to explain in vivo phenotypes using these data. A simple mathematical model describing the impact these catalytic deficiencies had on 2AA production was generally supported by our data. However, caveats arose when kinetic parameters, determined in vitro, were used to predict formation of the isoleucine precursor 2-ketobutyrate and model in vivo (growth) behaviors. Altogether, our data support the need for a holistic approach, including in vivo and in vitro analyses, to generate data used in understanding and modeling metabolism.

Keywords: 2-aminoacrylate; RidA; Salmonella enterica; bacterial metabolism; mathematical modeling; metabolic network; pyridoxal phosphate; reactive nitrogen species (RNS); serine/threonine dehydratase; stress.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Acrylates / metabolism*
  • Alleles
  • Biocatalysis
  • Kinetics
  • L-Serine Dehydratase / genetics*
  • L-Serine Dehydratase / metabolism*
  • Mutation*
  • Salmonella enterica / enzymology*


  • Acrylates
  • L-Serine Dehydratase