Skip to main content
NCBI home page
Applied and Environmental Microbiology logoLink to Applied and Environmental Microbiology
. 1993 May;59(5):1376–1382. doi: 10.1128/aem.59.5.1376-1382.1993

Purification and Characterization of Alkaline Xylanases from Bacillus polymyxa

P Morales 1, A Madarro 1, J A Pérez-González 1,*, J M Sendra 1, F Piñaga 1, A Flors 1
PMCID: PMC182092  PMID: 16348932

Abstract

By applying different classical and fast protein liquid chromatographic techniques, three xylanases (β-1,4-d-xylan xylanhydrolase) were purified to homogeneity from the extracellular enzymatic complex of Bacillus polymyxa. The three enzymes (X34C, X34E, and X22) were small proteins of 34, 34, and 22 kDa and basic pIs 9.3, >9.3, and 9.0, respectively. X34C and X34E are closely related and seem to be isoforms of the same enzyme. However, they differ in some characteristics. The three enzymes had different pH and temperature optima. One of them, X34E, showed a high thermal stability. The Vmax values determined for X34C, X34E, and X22 enzymes on oat spelts xylan were 14.9, 85.5, and 64.0 U mg-1, respectively, and 16.1, 62.0, and 150.6 U mg-1 on birchwood xylan. When oat spelts xylan was the substrate used, Km values of 3.4, 2.4, and 1.9 mg ml-1 were obtained for X34C, X34E, and X22 enzymes, respectively, and 0.65, 6.3, and 0.32 mg ml-1 were the respective Km values determined with birchwood xylan as the substrate. The enzymes were nondebranching endo-β-xylanases. Xylose was one of the products of xylan hydrolysis by xylanases X34C and X34E, but this monosaccharide was not released by X22 enzyme. However, neither of the enzymes was able to degrade xylobiose.

Full text

PDF
1376

Images in this article

1378Image
on p.1378

Selected References

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

  1. Bachmann S. L., McCarthy A. J. Purification and Cooperative Activity of Enzymes Constituting the Xylan-Degrading System of Thermomonospora fusca. Appl Environ Microbiol. 1991 Aug;57(8):2121–2130. doi: 10.1128/aem.57.8.2121-2130.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bernier R., Desrochers M., Jurasek L., Paice M. G. Isolation and Characterization of a Xylanase from Bacillus subtilis. Appl Environ Microbiol. 1983 Aug;46(2):511–514. doi: 10.1128/aem.46.2.511-514.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1016/0003-2697(76)90527-3. [DOI] [PubMed] [Google Scholar]
  4. Dekker R. F., Richards G. N. Hemicellulases: their occurrence, purification, properties, and mode of action. Adv Carbohydr Chem Biochem. 1976;32:277–352. doi: 10.1016/s0065-2318(08)60339-x. [DOI] [PubMed] [Google Scholar]
  5. Flint H. J., McPherson C. A., Martin J. Expression of two xylanase genes from the rumen cellulolytic bacterium Ruminococcus flavefaciens 17 cloned in pUC13. J Gen Microbiol. 1991 Jan;137(1):123–129. doi: 10.1099/00221287-137-1-123. [DOI] [PubMed] [Google Scholar]
  6. Gosalbes M. J., Pérez-González J. A., González R., Navarro A. Two beta-glycanase genes are clustered in Bacillus polymyxa: molecular cloning, expression, and sequence analysis of genes encoding a xylanase and an endo-beta-(1,3)-(1,4)-glucanase. J Bacteriol. 1991 Dec;173(23):7705–7710. doi: 10.1128/jb.173.23.7705-7710.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Matte A., Forsberg C. W. Purification, characterization, and mode of action of endoxylanases 1 and 2 from Fibrobacter succinogenes S85. Appl Environ Microbiol. 1992 Jan;58(1):157–168. doi: 10.1128/aem.58.1.157-168.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Robson L. M., Chambliss G. H. Characterization of the cellulolytic activity of a Bacillus isolate. Appl Environ Microbiol. 1984 May;47(5):1039–1046. doi: 10.1128/aem.47.5.1039-1046.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. SMOGYI M. Notes on sugar determination. J Biol Chem. 1952 Mar;195(1):19–23. [PubMed] [Google Scholar]
  10. Wong K. K., Tan L. U., Saddler J. N. Multiplicity of beta-1,4-xylanase in microorganisms: functions and applications. Microbiol Rev. 1988 Sep;52(3):305–317. doi: 10.1128/mr.52.3.305-317.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Yang R. C., Mackenzie C. R., Bilous D., Seligy V. L., Narang S. A. Molecular Cloning and Expression of a Xylanase Gene from Bacillus polymyxa in Escherichia coli. Appl Environ Microbiol. 1988 Apr;54(4):1023–1029. doi: 10.1128/aem.54.4.1023-1029.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Zanini V., Viviani M. A., Cava L., Tortorano A. M., Villa G. Candida infections in the burn patients. Panminerva Med. 1983 Jul-Sep;25(3):163–166. [PubMed] [Google Scholar]

Articles from Applied and Environmental Microbiology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES