1887

Abstract

This study has shown that strain K21 differs from the previously characterized and closely related strain PAP996 in that expression of the pullulanase gene () and other genes of the maltose regulon is partially independent of exogenous inducer (maltose/maltotriose). Mutants of strain K21 which are defective in pullulanase synthesis and/or secretion were isolated following Tn mutagenesis. Three phenotypic classes of mutants were identified. Class I mutants were defective in the surface localization and secretion of pullulanase. Class II mutants did not secrete detectable levels of pullulanase but were able to export pullulanase to the cell surface. Class II mutants also expressed pullulanase and other maltose-regulated genes at markedly lower levels than those found in the parent strain under non-inducing conditions. The single class III mutant was intermediate between K21 and class I mutants; most of the cell-associated pullulanase was localized at the cell surface whilst a significant amount was secreted into the medium. Mapping indicated that all but three of the Tn insertions were adjacent to, and at either side of, . One class II mutant carried a Tn insertion in or close to whereas in the remaining class II mutants the insertions were located at least 4 kb upstream of in a region which may define a new regulatory locus of the maltose operon.

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1989-02-01
2021-10-19
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References

  1. Bender H., Wallenfels K. 1966; Pullulanase (an amylopectin and glucogen debranching enzyme) from Aerobacter aerogenes . Methods in Enzymology 8:555–559
    [Google Scholar]
  2. Bloch M.A., Raibaud O. 1986a; Comparison of the malA regions of Escherichia coli and Klebsiella pneumoniae . Journal of Bacteriology 168:1220–1227
    [Google Scholar]
  3. Bloch M.A., Raibaud O. 1986b; Caracterisation preliminaire d’un nouveau systeme permettant l’assimilation du maltose par Escherichia coli . Annales de l’Institut Pasteur/Microbiologie 137:145–153
    [Google Scholar]
  4. Chang A.C.Y., Cohen S.N. 1978; Construction and characterisation of amplifiable multicopy DNA cloning vehicles derived from the P15A cryptic miniplasmid. Journal of Bacteriology 134:1141–1156
    [Google Scholar]
  5. Chapon C. 1982; Role of the catabolite activator protein in the maltose regulon of E. coli . Journal of Bacteriology 150:722–729
    [Google Scholar]
  6. Chapon C., Raibaud O. 1985; Structure of two divergent promoters located in front of the gene encoding pullulanase in Klebsiella pneumoniae and positively regulated by the malT product. Journal of Bacteriology 164:639–645
    [Google Scholar]
  7. Churchward G.G., Holland I.B. 1976; Envelope synthesis during the cell cycle in E. coli . Journal of Molecular Biology 105:245–261
    [Google Scholar]
  8. Clement J.M., Perrin D., Hedgepeth J. 1982; Analysis of λ receptor and β-lactamase synthesis and export using cloned genes in a minicell system. Molecular and General Genetics 185:302–310
    [Google Scholar]
  9. Cole S.T., Raibaud O. 1986; The nucleotide sequence of the malT gene encoding the positive regulator of the Escherichia coli maltose regulon. Gene 42:201–208
    [Google Scholar]
  10. Ehrmann M., Boos W. 1987; Identification of endogenous inducers of the mal regulon in Escherichia coli . Journal of Bacteriology 169:3539–3545
    [Google Scholar]
  11. D’Enfert C., Chapon C., Pugsley A.P. 1987a; Export and secretion of the lipoprotein pullulanase by Klebsiella pneumoniae . Molecular Microbiology 1:107–116
    [Google Scholar]
  12. D’Enfert C., Ryter A., Pugsley A.P. 1987b; Cloning and expression in E. coli of the Klebsiella pneumoniae genes for production, surface localisation and secretion of the lipoprotein pullulanase. EMBO Journal 6:3531–3538
    [Google Scholar]
  13. D’Enfert , Reyss I., Ryter A., Pugsley A.P. 1988; Pullulanase: a new specific secretion pathway in Escherichia coli . In Membrane Biochemistry pp. 419–428 Op den Kamp J. Edited by Berlin & Heidelberg: Springer Verlag;
    [Google Scholar]
  14. Feinberg A.P., Vogelstein B. 1983; A technique for radiolabelling DNA restriction endonuclease fragments to high specific activity. Analytical Biochemistry 132:6–13
    [Google Scholar]
  15. Filip C., Fletcher G., Wulff J.L., Earhart C. 1973; Solubilisation of the cytoplasmic membrane of Escherichia coli by an ionic detergent sodium lauryl sarcosinate. Journal of Bacteriology 115:717–722
    [Google Scholar]
  16. Freundlieb S., Boos W. 1986; α-Amylase of E. coli, mapping and cloning of the structural gene, malS, and identification of its product as a periplasmic protein. Journal of Biological Chemistry 261:2946–2953
    [Google Scholar]
  17. Gilson E., Rousset J.P., Charbit A., Perrin D., Hofnung M. 1986; malM, a new gene of the maltose regulon in Escherichia coli K-12. Journal of Molecular Biology 191:303–311
    [Google Scholar]
  18. Guidi-Rontani C., Gicquel-Sanzey B. 1981; Expression of the maltose regulon in strains lacking the cyclic AMP-receptor protein. FEMS Microbiological tetters 10:383–387
    [Google Scholar]
  19. Hope G.C., Dean A.C.R. 1974; Pullulanase synthesis in Klebsiella (Aerobacter) aerogenes strains growing in continuous culture. Biochemical Journal 144:403–411
    [Google Scholar]
  20. Katsuragi N., Takizawa N., Murooka Y. 1987; Entire nucleotide sequence of the pullulanase gene of Klebsiella aerogenes W70. Journal of Bacteriology 169:2301–2306
    [Google Scholar]
  21. Konishi Y., Amemura A., Tanabe S., Harada T. 1979; Immunological study of pullulanase from Klebsiella strains and the occurrence of this enzyme in the Enterobacteriaceae . International Journal of Systematic Bacteriology 29:13–18
    [Google Scholar]
  22. Kornacker M.G., Boyd A., Plastow G.S. 1988; A genetic analysis of pullulanase export from Klebsiella aerogenes . In Membrane Biochemistry pp. 429–438 Op den Kamp J. Edited by Berlin & Heidelberg: Springer Verlag;
    [Google Scholar]
  23. Kuhn I., Stephenson F., Boyer H.W., Green P.J. 1986; Positive-selection vectors utilizing lethality of the EcoKl endonuclease. Gene 44:253–263
    [Google Scholar]
  24. Laemmli U.K. 1970; Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature, tendon 227:680–685
    [Google Scholar]
  25. Maniatis T., Fritsch E.F., Sambrook J. 1982 Molecular Cloning, a Laboratory Manual Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
    [Google Scholar]
  26. Michaelis S.C., Chapon C., D’Enfert C., Pugsley A.P., Schwartz M. 1985; Characterisation and expression of the structural gene for pullulanase, a maltose-inducible secreted protein of Klebsiella pneumoniae . Journal of Bacteriology 164:633–638
    [Google Scholar]
  27. Miller J.H. 1972 Experiments in Molecular Genetics Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
    [Google Scholar]
  28. Morgan F.J., Adams K.R., Priest F.G. 1979; A cultural method for the detection of pullulan- degrading enzymes in bacteria and its application to the genus Bacillus . Journal of Applied Bacteriology 46:291–294
    [Google Scholar]
  29. Nilsson B., Uhlen M., Josephson S., Gatenbeck S., Philipson L. 1983; An improved positive selection plasmid vector constructed by oligonucleotide mediated mutagenesis. Nucleic Acids Research 11:8019–8030
    [Google Scholar]
  30. Plastow G.S., Border P.M., Hinton J.C.D., Salmond G.P.C. 1986; Molecular cloning of pectinase genes from Erwinia carotovora subspecies carotovora strain SCRI193. Symbiosis 2:115–122
    [Google Scholar]
  31. Pugsley A.P., Oudega B. 1987; Methods for studying colicins and their plasmids. In Plasmids, a Practical Approach pp. 105–160 Hardy K. G. Edited by Oxford: IRL Press;
    [Google Scholar]
  32. Pugsley A.P., Chapon C., Schwartz M. 1986; Extracellular pullulanase of Klebsiella pneumoniae is a lipoprotein. Journal of Bacteriology 166:1083–1088
    [Google Scholar]
  33. Raibaud O., Richet E. 1987; Maltotriose is the inducer of the maltose regulon of Escherichia coli . Journal of Bacteriology 169:3059–3061
    [Google Scholar]
  34. Raibaud O., Gutierrez C., Schwartz M. 1985; Essential and nonessential sequences in malPp, a positively controlled promoter in Escherichia coli . Journal of Bacteriology 161:1201–1208
    [Google Scholar]
  35. Rinderknecht H., Wilding P., Haverback P.J. 1967; A new method for the detection of α-amylase. Experientia 23:805–807
    [Google Scholar]
  36. Salmond G.P.C., Hinton J.C.D., Gill D.R., Perombelon M.C.M. 1986; Transposon mutagenesis of Erwinia using phage λ vectors. Molecular and General Genetics 203:524–528
    [Google Scholar]
  37. Schwartz M. 1987; The maltose regulon. In Escherichia coli and Salmonella typhimurium, Cellular and Molecular Biology pp. 1482–1502 Neidhardt F. C., Ingraham J. L., Low K. B., Magasanik B., Schaechter M., Umbarger H. E. Edited by Washington, DC: American Society for Microbiology;
    [Google Scholar]
  38. Takizawa N., Murooka Y. 1985; Cloning of the pullulanase gene and overproduction of pullulanase in Escherichia coli and Klebsiella aerogenes . Applied and Environmental Microbiology 49:294–298
    [Google Scholar]
  39. Way J.C., Davis M.A., Morisato D., Roberts D.E., Kleckner N. 1984; New Tn10 derivatives for transposon mutagenesis and for construction of lacZ operon fusions by transposition. Gene 32:369–379
    [Google Scholar]
  40. Woehner G., Woeber G. 1978a; Pullulanase, an enzyme of starch catabolism, is associated with the outer membrane of Klebsiella . Archives of Microbiology 116:303–310
    [Google Scholar]
  41. Woehner G., Woeber G. 1978b; Subcellular distribution of enzymes involved in α-glucan utilisation in Klebsiella pneumoniae . Archives of Microbiology 116:311–316
    [Google Scholar]
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