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. 1984 Nov;160(2):687–690. doi: 10.1128/jb.160.2.687-690.1984

Proton suicide: general method for direct selection of sugar transport- and fermentation-defective mutants.

J W Winkelman, D P Clark
PMCID: PMC214790  PMID: 6094484

Abstract

We devised a positive selection procedure for bacterial mutants incapable of producing acid from sugars by fermentation. The method relied on the production of elemental bromine from a mixture of bromide and bromate under acidic conditions. When wild-type Escherichia coli cells were plated on media containing a fermentable sugar and an equimolar mixture of bromide and bromate, most of the cells were killed but a variety of mutants unable to produce acid from the sugar survived. Among these mutants were those defective in (i) sugar uptake, (ii) the glycolytic pathway, and (iii) the excretion. There were also novel mutants with some presumed regulatory defects affecting fermentation.

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Selected References

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  1. Anderson A., Cooper R. A. Genetic mapping of a locus for triosephosphate isomerase on the genome of Escherichia coli K12. J Gen Microbiol. 1970 Aug;62(3):329–334. doi: 10.1099/00221287-62-3-329. [DOI] [PubMed] [Google Scholar]
  2. Bachmann B. J. Linkage map of Escherichia coli K-12, edition 7. Microbiol Rev. 1983 Jun;47(2):180–230. doi: 10.1128/mr.47.2.180-230.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Brice C. B., Kornberg H. L. Location on the chromosome of Escherichia coli of a gene specifying phosphopyruvate synthase activity. Biochim Biophys Acta. 1967 Mar 22;136(2):412–414. doi: 10.1016/0304-4165(67)90094-3. [DOI] [PubMed] [Google Scholar]
  4. Brown T. D., Jones-Mortimer M. C., Kornberg H. L. The enzymic interconversion of acetate and acetyl-coenzyme A in Escherichia coli. J Gen Microbiol. 1977 Oct;102(2):327–336. doi: 10.1099/00221287-102-2-327. [DOI] [PubMed] [Google Scholar]
  5. Courtright J. B., Henning U. Malate dehydrogenase mutants in Escherichia coli K-12. J Bacteriol. 1970 Jun;102(3):722–728. doi: 10.1128/jb.102.3.722-728.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Creaghan I. T., Guest J. R. Succinate dehydrogenase-dependent nutritional requirement for succinate in mutants of Escherichia coli K12. J Gen Microbiol. 1978 Jul;107(1):1–13. doi: 10.1099/00221287-107-1-1. [DOI] [PubMed] [Google Scholar]
  7. Guest J. R. Anaerobic growth of Escherichia coli K12 with fumarate as terminal electron acceptor. Genetic studies with menaquinone and fluoroacetate-resistant mutants. J Gen Microbiol. 1979 Dec;115(2):259–271. doi: 10.1099/00221287-115-2-259. [DOI] [PubMed] [Google Scholar]
  8. Hillman J. D., Fraenkel D. G. Glyceraldehyde 3-phosphate dehydrogenase mutants of Escherichia coli. J Bacteriol. 1975 Jun;122(3):1175–1179. doi: 10.1128/jb.122.3.1175-1179.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Irani M. H., Maitra P. K. Glyceraldehyde 3-p dehydrogenase, glycerate 3-P kinase and enolase mutants of Escherichia coli: genetic studies. Mol Gen Genet. 1976 Apr 23;145(1):65–71. doi: 10.1007/BF00331559. [DOI] [PubMed] [Google Scholar]
  10. Irani M., Maitra P. K. Isolation and characterization of Escherichia coli mutants defective in enzymes of glycolysis. Biochem Biophys Res Commun. 1974 Jan;56(1):127–133. doi: 10.1016/s0006-291x(74)80324-4. [DOI] [PubMed] [Google Scholar]
  11. Kline E. S., Mahler H. R. The lactic dehydrogenases of E. coli. Ann N Y Acad Sci. 1965 Jul 31;119(3):905–919. doi: 10.1111/j.1749-6632.1965.tb47451.x. [DOI] [PubMed] [Google Scholar]
  12. LeVine S. M., Ardeshir F., Ames G. F. Isolation and Characterization of acetate kinase and phosphotransacetylase mutants of Escherichia coli and Salmonella typhimurium. J Bacteriol. 1980 Aug;143(2):1081–1085. doi: 10.1128/jb.143.2.1081-1085.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Lorowitz W., Clark D. Escherichia coli mutants with a temperature-sensitive alcohol dehydrogenase. J Bacteriol. 1982 Nov;152(2):935–938. doi: 10.1128/jb.152.2.935-938.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Maloy S. R., Bohlander M., Nunn W. D. Elevated levels of glyoxylate shunt enzymes in Escherichia coli strains constitutive for fatty acid degradation. J Bacteriol. 1980 Aug;143(2):720–725. doi: 10.1128/jb.143.2.720-725.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Postma P. W., Roseman S. The bacterial phosphoenolpyruvate: sugar phosphotransferase system. Biochim Biophys Acta. 1976 Dec 14;457(3-4):213–257. doi: 10.1016/0304-4157(76)90001-0. [DOI] [PubMed] [Google Scholar]
  16. Roehl R. A., Vinopal R. T. Genetic locus, distant from ptsM, affecting enzyme IIA/IIB function in Escherichia coli K-12. J Bacteriol. 1980 Apr;142(1):120–130. doi: 10.1128/jb.142.1.120-130.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Saier M. H., Jr Bacterial phosphoenolpyruvate: sugar phosphotransferase systems: structural, functional, and evolutionary interrelationships. Bacteriol Rev. 1977 Dec;41(4):856–871. doi: 10.1128/br.41.4.856-871.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Shaw L., Grau F., Kaback H. R., Hong J. S., Walsh C. Vinylglycolate resistance in Escherichia coli. J Bacteriol. 1975 Mar;121(3):1047–1055. doi: 10.1128/jb.121.3.1047-1055.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Tarmy E. M., Kaplan N. O. Kinetics of Escherichia coli B D-lactate dehydrogenase and evidence for pyruvate-controlled change in conformation. J Biol Chem. 1968 May 25;243(10):2587–2596. [PubMed] [Google Scholar]
  20. VOGEL H. J., BONNER D. M. Acetylornithinase of Escherichia coli: partial purification and some properties. J Biol Chem. 1956 Jan;218(1):97–106. [PubMed] [Google Scholar]

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