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Antonie van Leeuwenhoek

, Volume 69, Issue 3, pp 235–241 | Cite as

Purification and characterization of two different xylanases from the thermophilic actinomycete Microtetraspora flexuosa SIIX

  • Stephan Berens
  • Herwig Kaspari
  • Jobst-Heinrich Klemme
Article

Abstract

Two endoxylanases were isolated from the xylanolytic enzyme system of the thermophilic actinomycete Microtetraspora flexuosa SIIX, and purified by ammonium sulfate fractionation, DEAE-Sepharose chromatography, gel filtration on Sephacryl S 200 and fast protein liquid chromatography on Q-Sepharose. The molecular masses of xylanase I and II were 26.3 and 16.8 kDa, and isoelectric points were 8.4 and 9.45, respectively. optimal enzyme activities were obtained at 80° C and pH 6.0. The thermostability of both xylanases was greatly diminished during purification but could be restored by preincubation of the purified enzymes in the presence of xylan. The half-lives at 80° C were approximately 25 min. The kinetic constants of xylanases I and II determined with Remazol-brilliant-blue xylan were Vmax of 1537 and 353 μmol·min-1·mg protein-1 and K m values of 2.44 and 1.07 mg·ml-1, respectively. Purified xylanases utilized xylan as well as small oligosaccharides such as xylotriose as substrate. They did not exhibit xylobiase or debranching activities. The predominant products of arabinoxylan hydrolysis were xylobiose and xylotriose, the latter being hydrolysed to xylobiose and xylose upon further incubation. In addition, fragments containing arabinose side chains accumulated. The xylanases did not act on crystalline or amorphous cellulose indicating a possible application in biobleaching processes.

Key words

Microtetraspora thermophilic actinomycetes xylanases 

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Copyright information

© Kluwer Academic Publishers 1996

Authors and Affiliations

  • Stephan Berens
    • 1
  • Herwig Kaspari
    • 1
  • Jobst-Heinrich Klemme
    • 1
  1. 1.Institut für Mikrobiologie und Biotechnologie der Universität BonnBonnGermany

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