1887

Abstract

Supernatants from cultures of contained up to 10 components with CM-cellulase activity as determined by non-denaturing PAGE. Some of the active components were glycosylated. The activity profiles of the supernatants, as determined by PAGE, varied with the cellulosic substrate used for the growth of a culture, with culture age, and with storage of the supernatants. These variations were a consequence of proteolysis and a reduction in the glycosylation of some of the components. Proteinase activity in the supernatants was induced by growth of on cellulosic substrates. Proteolysis and a reduction in glycosylation resulted in the conversion of slow-moving into fast-moving components on non-denaturing PAGE. The fast-moving components had a reduced ability to bind to an insoluble cellulosic substrate such as Avicel. Several of the CM-cellulase activities in culture supernatants were immunologically related. In contrast to the large number of CM-cellulases found in the supernatant, substrate-bound activity comprised only three slow-moving components, at least some of which were glycosylated. It was concluded that the cellulase system of was composed of only three enzymes, and that these enzymes had a great affinity for, and were stabilized by binding to, an insoluble cellulosic substrate. Enzymes which accumulated free in the culture medium were subject to limited proteolysis and de-glycosylation which generated a variety of products, some of which retained enzymic activity.

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1984-06-01
2021-10-20
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References

  1. Beguin P., Eisen H. 1978; Purification and partial characterization of three extracellular cellulases from Cellulomonas sp. European Journal of Biochemistry 87:525–531
    [Google Scholar]
  2. Berghem L. E. R, Pettersson L. G., Axio-Fredricksson U.-B. 1976; The mechanism of enzymatic cellulose degradation. Purification and some properties of two different l,4-β-glucan glucanohydrolases from Trichoderma viride. . European Journal of Biochemistry 61:621–630
    [Google Scholar]
  3. Choi W. Y., Haggett K. D., Dunn N. W. 1978; Isolation of a cotton wool degrading strain of Cellulomonas: mutants with altered ability to degrade cotton wool. Australian Journal of Biological science 31:553–564
    [Google Scholar]
  4. Choudhury N., Gray P. P., Dunn N. W. 1980; Saccharification of sugar cane bagasse by an enzyme preparation from Cellulomonas: resistance to product inhibition. Biotechnology tetters 2:427–428
    [Google Scholar]
  5. Enger M. D., Sleeper B. P. 1965; Multiple cellulase system from Streptomyces antibioticus. . Journal of Bacteriology 89:23–27
    [Google Scholar]
  6. Gilkes N. R., Kilburn D. G., Langsford M. L., Miller R. C. Jr Wakarchuk W. W., Warren R. A. J, Whittle D. J. 1984; Isolation and characterization of Escherichia coli clones expressing cellulase genes from Cellulomonas ftmi. . Journal of General Microbiology 130: in the Press
    [Google Scholar]
  7. Gum E. K., Brown R. D. 1977; Comparison of four purified extracellular l,4-β-d-glucan cellobio- hydrolase enzymes from Trichoderma viride. . Biochi-mica et biophysica acta 492:225–231
    [Google Scholar]
  8. Haggett K. D., Gray P. P., Dunn N. W. 1979; Crystalline cellulose degradation by a strain of Cellulomonas and its mutant derivatives. European Journal of Applied Microbiology and Biotechnology 8:183–190
    [Google Scholar]
  9. Jovin T., Chrambach A., Naughton M. A. 1964; An apparatus for temperature-regulated polyacrylamide gel electrophoresis. Analytical Biochemistry 9:351–369
    [Google Scholar]
  10. Laemmli U. K. 1970; Cleavage of structural proteins during assembly of the head of bacteriophage T4. Nature; London: 277680–685
    [Google Scholar]
  11. Mcguckin W. F., Mckenzie B. F. 1958; An improved periodic acid fuchsin sulfite staining method for evaluation of glycoproteins. Clinical Chemistry 4:476–483
    [Google Scholar]
  12. Miller G. L. 1959; Use of dinitrosalicylic acid reagent for determination of reducing sugars. Analytical Chemistry 31:426–428
    [Google Scholar]
  13. Millett M. A., Baker A. J., Satter L. D. 1976; Physical and chemical pretreatments for enhancing cellulose saccharification. Biotechnology and Bioengineering Symposium 6:125–153
    [Google Scholar]
  14. O’Farrell P. H. 1975; High resolution two-dimensional electrophoresis of proteins. Journal of Biological Chemistry 250:4007–4021
    [Google Scholar]
  15. Ol V. T., Herzenberg L. 1980; Immunoglobulin- producing hybrid cell lines. In Selected Methods in Cellular Immunology pp. 351–372 Mishell B. B., Shiigi S. M. Edited by San Francisco: W. H. Freeman;
    [Google Scholar]
  16. Rinderknecht H., Geokas M. C., Silverman P., Haverback B. J. 1968; A new ultrasensitive method for the determination of proteolytic activity. Clinica chimica acta 21:197–203
    [Google Scholar]
  17. Saddler J. N., Khan A. W. 1981; Cellulolytic enzyme system of Acetivibrio cellulolyticus. . Canadian Journal of Microbiology 27:288–294
    [Google Scholar]
  18. schleif R. F., Wensink P. C. 1981 Practical Methods in Molecular Biology pp. xiii + 220 New York: Springer-Verlag;
    [Google Scholar]
  19. schneider C., Newman R. A., Sutherland D. R., Asser U., Greaves M. F. 1982; A one-step purification of membrane proteins using a high efficiency immunomatrix. Journal of Biological Chemistry 257:10766–10769
    [Google Scholar]
  20. Stewart B. J., Leatherwood J. M. 1976; Derepressed synthesis of cellulase by Cellulomonas. . Journal of Bacteriology 128:609–615
    [Google Scholar]
  21. Whittle D. J., Kilburn D. G., Warren R. A. J, Miller R. C. Jr 1982; Molecular cloning of a Cellulomonas fimi cellulase gene in Escherichia coli. . Gene 17:139–145
    [Google Scholar]
  22. Yamane K., Suzuki H., Nisizawa K. 1970; Purification and properties of extracellular and cell- bound cellulase components of Pseudomonas fluores- cens var.cellulosa. . Journal of Biochemistry 67:19–35
    [Google Scholar]
  23. Zacharius R. M., Zell T. E., Morrison J. H. 1969; Glycoprotein staining following electrophoresis on acrylamide gels. Analytical Biochemistry 30:148–152
    [Google Scholar]
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