Formate dehydrogenase mutants of Salmonella typhimurium: a new medium for their isolation and new mutant classes.
We have designed a new medium for the differentiation of mutants of Salmonella typhimurium defective in the ability to reduce nitrate with formate, and have characterized 24 formate dehydrogenase (FDH) mutants isolated on this medium. The mutants were assayed for the ability to use formate to reduce benzyl viologen and phenazine methosulfate, and were mapped by means of conjugation and P22-mediated transduction. Mutants lacking the ability to reduce either dye were found to map at three distinct sites: at a site co-transducible with xyl (presumably fdhA), at a site or sites between 13U and 33U, but not co-transducible with aroA, bio, purB, pyrC, or pyrD (near, but not identical with fdhB), and at asite 10-20% co-transducible with pyrE, for which we suggest the designation fdhC. Six mutant isolates reduced benzyl viologen, but not phenazine methosulfate. They retained the ability to produce nitrite during growth with nitrate. They mapped between 83U and 89U, but no co-transduction was found with metE, glnA, metB, or argH. The combined biochemical and genetic data suggest the existence of a gene in this area which is essential for the reduction of nitrate with formate, but not for formate hydrogenlyase activity or for nitrate reductase activity.
D000445:Aldehyde Oxidoreductases / Q000235:genetics*; D001594:Benzyl Viologen / Q000378:metabolism; D002874:Chromosome Mapping; D002876:Chromosomes, Bacterial / Q000648:ultrastructure; D003227:Conjugation, Genetic; D003470:Culture Media; D005560:Formate Dehydrogenases / Q000235:genetics*; D005561:Formates; D008773:Methylphenazonium Methosulfate / Q000378:metabolism; D009154:Mutation; D009566:Nitrates / Q000378:metabolism; D012486:Salmonella typhimurium / Q000201:enzymology / Q000235:genetics*; D014161:Transduction, Genetic
D003470:Culture Media; D005561:Formates; D009566:Nitrates; D001594:Benzyl Viologen; D008773:Methylphenazonium Methosulfate; D005560:Formate Dehydrogenases; D000445:Aldehyde Oxidoreductases