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. 1988 Nov;54(11):2881–2883. doi: 10.1128/aem.54.11.2881-2883.1988

Purification and Characterization of Thermostable Pullulanase from Bacillus stearothermophilus and Molecular Cloning and Expression of the Gene in Bacillus subtilis

Takashi Kuriki 1, Jong-Hyun Park 1, Shigetaka Okada 1, Tadayuki Imanaka 1,*
PMCID: PMC204393  PMID: 16347785

Abstract

A thermostable pullulanase (α-dextrin 6-glucanohydrolase [EC 3.2.1.41]) from a newly isolated Bacillus stearothermophilus strain (TRS128) was purified and characterized. The enzyme hydrolyzed (1→6)-α-d-glucosidic linkages of pullulan to produce maltotriose, and the optimum temperature was 65°C. About 90% of the enzyme activity was retained after treatment at 65°C for 60 min. By using pTB522 as a vector plasmid, the pullulanase gene was cloned and expressed in Bacillus subtilis.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Aiba S., Kitai K., Imanaka T. Cloning and Expression of Thermostable alpha-Amylase Gene from Bacillus stearothermophilus in Bacillus stearothermophilus and Bacillus subtilis. Appl Environ Microbiol. 1983 Nov;46(5):1059–1065. doi: 10.1128/aem.46.5.1059-1065.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Coleman R. D., Yang S. S., McAlister M. P. Cloning of the debranching-enzyme gene from Thermoanaerobium brockii into Escherichia coli and Bacillus subtilis. J Bacteriol. 1987 Sep;169(9):4302–4307. doi: 10.1128/jb.169.9.4302-4307.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Eisele B., Rasched I. R., Wallenfels K. Molecular characterization of pullulanase from Aerobacter aerogenes. Eur J Biochem. 1972 Mar 15;26(1):62–67. doi: 10.1111/j.1432-1033.1972.tb01739.x. [DOI] [PubMed] [Google Scholar]
  4. Hyun H. H., Zeikus J. G. General Biochemical Characterization of Thermostable Pullulanase and Glucoamylase from Clostridium thermohydrosulfuricum. Appl Environ Microbiol. 1985 May;49(5):1168–1173. doi: 10.1128/aem.49.5.1168-1173.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Imanaka T., Himeno T., Aiba S. Effect of in vitro DNA rearrangement in the NH2-terminal region of the penicillinase gene from Bacillus licheniformis on the mode of expression in Bacillus subtilis. J Gen Microbiol. 1985 Jul;131(7):1753–1763. doi: 10.1099/00221287-131-7-1753. [DOI] [PubMed] [Google Scholar]
  6. Katsuragi N., Takizawa N., Murooka Y. Entire nucleotide sequence of the pullulanase gene of Klebsiella aerogenes W70. J Bacteriol. 1987 May;169(5):2301–2306. doi: 10.1128/jb.169.5.2301-2306.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Kuriki T., Okada S., Imanaka T. New type of pullulanase from Bacillus stearothermophilus and molecular cloning and expression of the gene in Bacillus subtilis. J Bacteriol. 1988 Apr;170(4):1554–1559. doi: 10.1128/jb.170.4.1554-1559.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Mercier C., Frantz B. M., Whelan W. J. An improved purification of cell-bound pullulanase from Aerobacter aerogenes. Eur J Biochem. 1972 Mar 15;26(1):1–9. doi: 10.1111/j.1432-1033.1972.tb01733.x. [DOI] [PubMed] [Google Scholar]
  9. Morgan F. J., Adams K. R., Priest F. G. A cultural method for the detection of pullulan--degrading enzymes in bacteria and its application to the genus Bacillus. J Appl Bacteriol. 1979 Apr;46(2):291–294. doi: 10.1111/j.1365-2672.1979.tb00823.x. [DOI] [PubMed] [Google Scholar]
  10. Plant A. R., Clemens R. M., Morgan H. W., Daniel R. M. Active-site- and substrate-specificity of Thermoanaerobium Tok6-B1 pullulanase. Biochem J. 1987 Sep 1;246(2):537–541. doi: 10.1042/bj2460537. [DOI] [PMC free article] [PubMed] [Google Scholar]

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