1887

Abstract

Xylan-degrading enzymes, including -xylosidase (EC 3.2.1.37), were induced when was grown at 50 °C in liquid medium containing 0·2% xylan. The intracellular -xylosidase activity was concentrated and characterized by fast protein liquid chromatography and gel electrophoresis. A zymogram technique was developed to identify -xylosidase directly on polyacrylamide gels. A single enzyme (168 kDa; pI 4·37) was identified and purified to homogeneity. The consistent detection of a single band on denaturing SDS gels suggested that the enzyme was composed of identical subunits; since the subunit molecular mass was 56 kDa, a trimeric structure is suggested. High activity against -nitrophenyl --xylopyranoside (pNPX) occurred in the pH range 5·0–9·0 and temperature range 40–60 °C. The enzyme was stable at room temperature at pH 6·0–8·0; it had a half-life of 8 h at 65 °C, and of 1·5 h at 70 °C. The purified enzyme did not exhibit any detectable activity against arabinoxylan, carboxymethylcellulose or -nitrophenyl --glucopyranoside. The enzyme had a of 0·89 m (pNPX) and was inhibited by -xylose ( 19 m) but not -glucose. The size of the enzyme is in the range reported for the few other bacterial -xylosidases described, but the acidic nature of the protein and its affinity for the substrate have more in common with some of the monomeric -xylosidases described in fungi.

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1989-02-01
2022-01-17
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References

  1. Biely P. 1985; Microbial xylanolytic systems. Trends in Biotechnology 3:286–290
    [Google Scholar]
  2. Biely P., Mackenzie C.R., Puls J., Schneider H. 1986; Cooperativity of esterases and xylanases in the enzymatic degradation of acetyl xylan. Biotechnology 4:731–733
    [Google Scholar]
  3. Calza R.E., Irwin D.C., Wilson D.B. 1985; Purification and characterisation of two ´-1,4-endoglucanases from Thermomonospora fusca . Biochemistry 24:7797–7804
    [Google Scholar]
  4. Chesson A., Gordon A.H., Lomax J.A. 1983; Substituent groups linked by alkali labile bonds to arabinose and xylose residues of legume, grass and cereal straw walls and their fate during digestion by rumen microorganisms. Journal of the Science of Food and Agriculture 34:1330–1340
    [Google Scholar]
  5. Claeyssens M., Saman E., Kersters-Hilderson H., De Bruyne C.K. 1975; β-d-Xylosidase from Bacillus pumilus: molecular properties and oligomeric structure. Biochimica et biophysica acta 405:475–481
    [Google Scholar]
  6. Dekker R.F.H., Richards G.N. 1976; Hemicellulases: their occurrence, properties and mode of action. In Advances in Carbohydrate Chemistry and Biochemistry, pp 278–352 Tipson R. S., Horton D. Edited by London: Academic Press;
    [Google Scholar]
  7. Deleyn F., Claeyssens M., Van Beelmen J., Debruyne C.K. 1978; Purification and properties of β-xylosidase from Penicillium wortmanni . Canadian Journal of Biochemistry 56:43–50
    [Google Scholar]
  8. Deshpande V., Lachke A., Mishra C., Keskar S., Rao M. 1986; Mode of action and properties of xylanase and β-xylosidase from Neurospora crassa . Biotechnology and Bioengineering 28:1832–1837
    [Google Scholar]
  9. John M., Schmidt B., Schmidt J. 1979; Purification and some properties of five endo-l,4,β-d-xylanases and a β-d-xylosidase produced a strain of Aspergillus niger . Canadian Journal of Biochemistry 57:125–134
    [Google Scholar]
  10. Johnson W.C., Lindsey A.J. 1939; An improved universal buffer. Analyst 64:490–492
    [Google Scholar]
  11. Kersters-Hilderson H., Loontiens F.G., Claeyssens M., Bruyne C.K. 1969; Partial purification and properties of an induced β-d-xylosidase of Bacillus pumilus 12. European Journal of Biochemistry 7:434–441
    [Google Scholar]
  12. Laemmli U.K. 1970; Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature, London 227:680–685
    [Google Scholar]
  13. Lee S.F., Forsberg C.W. 1987; Isolation and some properties of a β-d-xylosidase from Clostridium acetobutylicum ATCC824. Applied and Environmental Microbiology 53:651–654
    [Google Scholar]
  14. Mccarthy A.J. 1987; Lignocellulose-degrading actinomycetes. FEMS Microbiology Reviews 46:145–163
    [Google Scholar]
  15. Mccarthy A.J., Peace E., Broda P. 1985; Studies on the extracellular xylanase activities of some thermophilic actinomycetes. Applied Microbiology and Biotechnology 21:238–244
    [Google Scholar]
  16. Mackenzie C.R., Bilous D., Schneider H., Johnson K.G. 1987; Induction of cellulolytic and xylanolytic enzyme systems in Streptomyces sp. Applied and Environmental Microbiology 53:2835–2839
    [Google Scholar]
  17. Matsuo M., Yasui T. 1984; Purification and properties of β-d-xylosidase from Trichoderma viride . Agricultural and Biological Chemistry 48:1845–1852
    [Google Scholar]
  18. Merril C.R., Goldman D., Sedman S.A., Ebert M.H. 1981; Ultrasensitive stain for proteins in polyacrylamide gels shows regional variation in cerebrospinal fluid proteins. Science 211:1437–1438
    [Google Scholar]
  19. Panbangred W., Kawaguchi O., Tomita T., Shinmyo A., Okada H. 1984; Isolation of two β-xylosidase genes of Bacillus pumilus and comparison of their gene products. European Journal of Biochemistry 138:267–273
    [Google Scholar]
  20. Puls J., Schmidt O., Granzow C. 1987; Glucuronidase in two microbial xylanolytic systems. Enzyme and Microbial Technology 9:83–88
    [Google Scholar]
  21. Reilly P.J. 1981; Xylanases: structure and function. In Trends in the Biology of Fermentation for Fuels and Chemicals, pp 111–129 Hollaender A. E. Edited by New York: Plenum;
    [Google Scholar]
  22. Ristroph D.L., Humphrey A.E. 1985; The β-xylosidase of Thermomonospora . Biotechnology and Bioengineering 27:909–913
    [Google Scholar]
  23. Shewale J.G. 1982; β-Glucosidase: its role in cellulase synthesis and hydrolysis of cellulose. International Journal of Biochemistry 14:435–443
    [Google Scholar]
  24. Takenishi S., Tsuliska Y., Fukomoto J. 1973; Studies on hemicellulases. IV. Purification and properties of the β-xylosidase produced by Aspergillus niger van Tieghem. Journal of Biochemistry 73:335–343
    [Google Scholar]
  25. Timel T.E. 1967; Recent progress in the chemistry of wood hemicellulose. Wood Science and Technology 1:45–70
    [Google Scholar]
  26. Uziie M., Matsuo M., Yasui T. 1985; Possible identity of β-xylosidase and β-glucosidase of Chaetomium trilaterale . Agricultural and Biological Chemistry 49:1167–1173
    [Google Scholar]
  27. Whistler R.H.A., Richards E.L. 1970; Hemicellulose. In The Carbohydrates, pp 447–469 Pigman W., Horton D. Edited by New York: Academic Press;
    [Google Scholar]
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