Summary
A method for the positive selection of dadA mutants defective in Dolor-amino acid dehydrogenase has been devised. It consists in isolating mutants resistant to β-chroro-Dolor-alanine and screening for mutant colony color on a special agar medium. All 70 Escherichia coli K12 dadA mutants isolated either by this method or by other selection procedures map at a locus which is near to hemA and closely linked with dadR. Since some of the dadA mutants are thermosensitive in Dolor-methionine utilization in vivo and have thermolabile Dolor-amino acid dehydrogenase in vitro, it is proposed that the dadA gene codes for the enzyme structure. The broad substrate specificity, apparent membrane localization, inducibility by alanine, and repressibility by glucose strongly suggest that the Dolor-amino acid dehydrogenase coded by the dadA gene is a species variant of the enzyme described under the same name in Salmonella typhimurium. It may be identical or homologous with the enzymes described under the names alaninase, Dolor-alanine oxidase or Dolor-alanine dehydrogenase in E. coli K12 or B.
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Communicated by G.R. Fink
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Wild, J., Kłopotowski, T. Dolor-Amino acid dehydrogenase of Escherichia coli K12: Positive selection of mutants defective in enzyme activity and localization of the structural gene. Molec. Gen. Genet. 181, 373–378 (1981). https://doi.org/10.1007/BF00425614
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DOI: https://doi.org/10.1007/BF00425614