A Genetic Analysis of Pullulanase Export from Klebsiella Aerogenes

  • M. G. Kornacker
  • A. Boyd
  • G. S. Plastow
Conference paper
Part of the NATO ASI Series book series (volume 16)


The enzyme pullulanase (EC is produced by various genera of Gram-positive and Gram-negative bacteria. This enzyme is defined by its ability to hydrolyze α(1–6) bonds of α(1–4), α(1–6) glucans. Pullulanases of approximately 140 kd (Eisele et al. 1972) are produced and secreted by strains of the Gram-negative bacterium Klebsiella aerogenes. The amounts synthesized and proportions secreted are strain-dependent. In this report we summarize data obtained from the analysis of Klebsiella aerogenes strain K21. This strain secretes 50% of pullulanase made into the culture medium. The remaining 50% is located at the cell surface.


HindIII Fragment BglII Fragment Maltose Utilisation Lipoprotein Nature Signal Peptidase Cleavage Site 
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  1. Braun V (1975) Covalent lipoprotein from the outer membrane of E. coli. Biochim Biophys Acta 415: 335–377PubMedGoogle Scholar
  2. 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. J Bacteriol 164: 639–645PubMedGoogle Scholar
  3. Eisele B, Rasched IR, Wallenfels K (1972) Molecular characterisation of pullulanase from Aerobacter aerogenes. Eur J Biochem 26: 62–67PubMedCrossRefGoogle Scholar
  4. Ichihara S, Hussain M, Mizushima S (1981) Characterisation of new membrane lipoproteins and their precursors in E. coli. J Biol Chem 256: 3125–3129PubMedGoogle Scholar
  5. Kadonaga JT, Gautier AE, Straus DR, Charles AD, Edge MD, Knowles JR (1984) The role of the βlactamase signal sequence in the secretion of proteins by E. coli. J Biol Chem 259: 2149–2154PubMedGoogle Scholar
  6. Katsuragi N, Takizawa N, Murooka Y (1987) Entire nucleotide sequence of the pullulanase gene of Klebsiella aerogenes W70. J Bacteriol 169: 2301–2306PubMedGoogle Scholar
  7. Kuhn I, Stephenson F, Boyer HW, Greene PJ (1986) Positive-selection vectors utilizing lethality of EcoR1 endonuclease. Gene 44: 253–263CrossRefGoogle Scholar
  8. Michaelis S, Chapon C, D’Enfert C, Pugsley AP, Schwartz M (1985) Characterisation and expression of the structural gene for pullulanase, a maltose-inducible secreted protein of Klebsiella pneumoniae. J Bacteriol 164: 633–638Google Scholar
  9. Morgan FJ, Adams KR, Priest FG (1979) A cultural method for the detection of pullulan-degrading enzymes in bacteria and its application to the genus Bacillus. J Appl Bacteriol 46: 291–294PubMedCrossRefGoogle Scholar
  10. Nielsen JBK, Lampen JO (1982) Glyceride-cysteine lipoproteins and secretion by gram-positive bacteria. J Bacteriol 152: 315–322PubMedGoogle Scholar
  11. Nilsson B, Uhlen M, Josephson S, Gatenbeck S, Philipson L (1983) An improved positive-selection plasmid vector constructed by oligonucleotide mediated mutagenesis. Nucleic Acids Res 11: 8019–8030PubMedCrossRefGoogle Scholar
  12. Ohba R, Ueda S (1982) An inductive effector in the production of extracellular pullulanase by Aerobacter aerogenes. Agric Biol Chem 46: 2425–2431CrossRefGoogle Scholar
  13. Pugsley AP, Chapon C, Schwartz M (1986) Extracellular pullulanase of Klebsiella pneumoniae is a lipoprotein. J Bacteriol 166: 1083–1088.PubMedGoogle Scholar
  14. Salmond GPC, Hinton JCD, Gill DR, Perombelon MCM (1986) Transposon mutagenesis of Erwinia using phage λ vectors. Mol Gen Genet 203: 524–528CrossRefGoogle Scholar
  15. Takizawa N, Murooka Y (1985) Cloning of the pullulanase gene and overproduction of pullulanase in E. coli and Klebsiella aerogenes. Appl Environ Microbiol 49: 294–298PubMedGoogle Scholar
  16. Way JC, Davis MA, Morisato D, Roberts DE, Kleckner N (1984) New Tn1O derivatives for transposon mutagenesis and for construction of lacZ operon fusions by transposition. Gene 32: 369–379PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 1988

Authors and Affiliations

  • M. G. Kornacker
    • 1
  • A. Boyd
    • 2
  • G. S. Plastow
    • 1
  1. 1.The Leicester BiocentreUniversity of LeicesterLeicesterGreat Britain
  2. 2.Dept. of Biochemistry, Medical SchoolUniversity of EdinburghEdinburghUK

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