1887

Abstract

A cDNA expression library of the rumen fungus was made in . Cellulolytic clones were identified by screening on a medium containing carboxymethylcellulose. Restriction mapping and Southern hybridization analysis of selected clones revealed three distinct cellulase cDNAs, designated and . Studies on the substrate specificity showed that the enzyme encoded by had high activity towards amorphous and microcrystalline cellulose, while the and enzymes had relatively high activity on carboxymethylcellulose, with little activity on microcrystalline cellulose. Analysis of hydrolysis products from defined cellodextrins showed that the and enzymes hydrolysed β-1,4-glucosidic linkages randomly, whereas the enzyme cleaved cellotetraose to cellobiose, and cellopentaose to cellobiose and cellotriose. Cellobiose was also the only product detectable from hydrolysis of microcrystalline cellulose by the enzyme. Based on substrate specificity and catalytic mode, appears to encode a cellobiohydrolase, while and encode endoglucanases. Northern blot hybridization analysis showed that expression of the three cellulase transcripts in was induced by cellulose.

Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-138-7-1413
1992-07-01
2021-08-04
Loading full text...

Full text loading...

/deliver/fulltext/micro/138/7/mic-138-7-1413.html?itemId=/content/journal/micro/10.1099/00221287-138-7-1413&mimeType=html&fmt=ahah

References

  1. Azevedo M., de O., Felipe M. S. S., Astolfi-Filho S., Radford A. 1990; Cloning, sequencing and homologies of the cbh-l (exoglucanase) gene of Humicola grisea var. thermoidea. Journal of General Microbiology 136 2569 2576
    [Google Scholar]
  2. Bauchop T. 1981; The anaerobic fungi in rumen fiber digestion. Agriculture and Environment 6 339 348
    [Google Scholar]
  3. Beguin P. 1990; Molecular biology of cellulose degradation. Annual Review of Microbiology 44 219 248
    [Google Scholar]
  4. Borneman W. S., Akin D. E. 1990; Lignocellulose degradation by rumen fungi and bacteria: ultrastructure and cell wall degrading enzymes. In Microbial and Plant Opportunities to Improve Lignocellulose Utilization by Ruminants pp. 325 340 Edited by Akin D. E., Ljungdahl L. G., Wilson J. R., Harris P. J. New York: Elsevier;
    [Google Scholar]
  5. Calza R. E. 1991; Carbon source, cyclic nucleotide, and protein inhibitor effects on protein and cellulase secretions in Neocallimastix frontalis EB188. Current Microbiology 22 213 219
    [Google Scholar]
  6. Chen C. M., Gritzali M., Stafford D. W. 1987; Nucleotide sequence and deduced primary structure of cellobiohydrolase II from Trichoderma reesei. Biol. Technology 5 274 278
    [Google Scholar]
  7. Chomczynski P., Sacchi N. 1987; Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Analytical Biochemistry 162 156 159
    [Google Scholar]
  8. Claeyssens M., van Tilbeurgh H., Tomme P., Wood T. M., McRae S. I. 1989; Fungal cellulase systems: comparison of the specificities of the cellobiohydrolases isolated from Penicillium pinophilum and Trichoderma reesei. Biochemical Journal 261 819 825
    [Google Scholar]
  9. Dehority B. A. 1991; Cellulose degradation in ruminants. In Biosynthesis and Biodegradation of Cellulose pp. 327 351 Edited by Haigler C. H., Weimer P. J. New York: Marcel Dekker;
    [Google Scholar]
  10. El-Gogary S., Leite A., Crivellaro O., Eveleigh D. E., El-Dorry H. 1989; Mechanism by which cellulose triggers cellobiohydrolase I gene expression in Trichoderma reesei. Proceedings of the National Academy of Sciences of the United States of America 86 6138 6141
    [Google Scholar]
  11. Flint H. J., McPherson C. A., Martin J. 1991; Expression of two xylanase genes from the rumen cellulolytic bacterium Ruminococcus flavefaciens 17 cloned in pUC13. Journal of General Microbiology 137 123 129
    [Google Scholar]
  12. Gilkes N. R., Langsford M. L., Kilburn D. G., Miller R. C., Warren R. A. J. 1984; Mode of action and substrate specificities of cellulases from cloned bacterial genes. Journal of Biological Chemistry 259 10455 10459
    [Google Scholar]
  13. Gilkes N. R., Kilburn D. G., Miller R. C., Warren R. A. J. 1991; Bacterial cellulases. Bioresource Technology 36 21 35
    [Google Scholar]
  14. Goyal A., Ghosh B., Eveleigh D. 1991; Characteristics of fungal cellulases. Bioresource Technology 36 37 50
    [Google Scholar]
  15. Hazlewood G. P., Davidson K., Laurie J. I., Romaniec M. P. M., Gilbert H. J. 1990; Cloning and sequencing of the celA gene encoding endoglucanase A of Butyrivibrio fibrisolvens strain A46. Journal of General Microbiology 136 2089 2097
    [Google Scholar]
  16. Kanda T., Nisizawa K. 1988; Exocellulase of Irpex lacteus. Methods in Enzymology 160 403 408
    [Google Scholar]
  17. Kemp P., Lander D. J., Orpin C. G. 1984; The lipids of the rumen fungus Piromonas communis. Journal of General Microbiology 130 27 37
    [Google Scholar]
  18. Kyriacou A., Mackenzie C. R., Neufeld R. J. 1987; Detection and the characterization of the specific and non-specific endoglucanases of Trichoderma reesei: evidence demonstrating endoglucanase activity by cellobiohydrolase II. Enzyme and Microbial Technology 9 25 31
    [Google Scholar]
  19. Lake B. D., Goodwin H. J. 1976; Lipids. In Chromatographic and Electrophoretic Techniques vol. 1 , 4th edn.. pp. 345 366 Edited by Smith I., Seakins J. W. T. Bath: Pitman Press;
    [Google Scholar]
  20. Li X., Calza R. E. 1991; Fractionation of cellulases from the ruminal fungus Neocallimastix frontalis EB188. Applied and Environmental Microbiology 57 3331 3336
    [Google Scholar]
  21. Messner R., Kubicek C. P. 1991; Carbon source control of cellobiohydrolase I and II formation by Trichoderma reesei. Applied and Environmental Microbiology 57 630 635
    [Google Scholar]
  22. Morag E., Halevy I., Bayer E. A., Lamed R. 1991; Isolation and properties of a major cellobiohydrolase from the cellulosome of Clostridium thermocellum. Journal of Bacteriology 173 4155 4162
    [Google Scholar]
  23. Orpin C. G., Letcher A. 1979; Utilization of starch, cellulose, xylan and other hemicelluloses for growth by the rumen phycomy-cete Neocallimastix frontalis. Current Microbiology 3 121 124
    [Google Scholar]
  24. Orpin C. G., Munn E. A. 1986; Neocallimastix patriciarum sp. nov., a new member of the Neocallimasticaceae inhabiting the rumen of sheep. Transactions of the British Mycological Society 86 178 180
    [Google Scholar]
  25. Penttilä M. E., Lehtovaara P., Bailey M., Teeri T. T., Knowles J. K. C. 1988; Efficient secretion of two fungal cellobiohydrolases by Saccharomyces cerevisiae. Gene 63 103 112
    [Google Scholar]
  26. Phillips M. W., Gordon G. L. R. 1989; Growth characteristics on cellobiose of three different anaerobic fungi isolated from the ovine rumen. Applied and Environmental Microbiology 55 1695 1702
    [Google Scholar]
  27. Reymond P., Durand R., Hebraud M., Fevre M. 1991; Molecular cloning of genes from the rumen anaerobic fungus Neocallimastix frontalis: expression during hydrolase induction. FEMS Microbiology Letters 77 107 112
    [Google Scholar]
  28. Robson L. M., Chambliss G. H. 1989; Cellulases of bacterial origin. Enzyme and Microbial Technology 11 626 644
    [Google Scholar]
  29. Sambrook J., Fritsch E. F., Maniatis T. 1989 Molecular Cloning: a Laboratory Manual , 2nd edn.. Cold Spring Harbor, NY: New York: Cold Spring Harbor Laboratory;
    [Google Scholar]
  30. Schwarz W. H., Schimming S., Staudenbauer W. L. 1987; High-level expression of Clostridium thermocellum cellulase genes in Esherichia coli. Applied Microbiology and Biotechnology 27 50 56
    [Google Scholar]
  31. Shoemaker S., Schweickart V., Ladner M., Gelfand D., Kwok S., Myambo K., Innis M. 1983; Molecular cloning of exo-cellobiohydrolase I derived from Trichoderma reesei strain L27. Biol. Technology 1 691 696
    [Google Scholar]
  32. Sims P., James C., Broda P. 1988; The identification, molecular cloning and characterization of a gene from Phanerochaete chrysosporium that shows strong homology to the exo-cellobiohydrolase I gene of Trichoderma reesei. Gene 74 411 422
    [Google Scholar]
  33. Teather R. M., Wood P. J. 1982; Use of Congo Red–polysaccharide interactions in enumeration and characterization of cellulolytic bacteria from the bovine rumen. Applied and Environmental Microbiology 43 777 780
    [Google Scholar]
  34. Teeri T., Salovuori I., Knowles J. 1983; The molecular cloning of the major cellulase gene from Trichoderma reesei. Biol. Technology 1 696 699
    [Google Scholar]
  35. Webb E. C. 1984; Enzyme Nomenclature. Recommendations of the Nomenclature Committee of the International Union of Biochemistry pp. 320 321 New York: Academic Press;
    [Google Scholar]
  36. Williams A. G., Orpin C. G. 1987; Polysaccharide-degrading enzymes formed by three species of anaerobic rumen fungi grown on a range of carbohydrate substrates. Canadian Journal of Microbiology 33 418 426
    [Google Scholar]
  37. Wood T. M. 1989; Mechanisms of cellulose digestion by enzymes from aerobic and anaerobic fungi. In Enzyme Systems for Lignocellulose Degradation pp. 17 35 Edited by Coughlan M. London: Elsevier Applied Science;
    [Google Scholar]
  38. Wood T. M., McCrae S. I., Wilson C. A., Bhat K. M., Gow L. A. 1988; Aerobic and anaerobic fungal cellulases, with special reference to their mode of attack on crystalline cellulose. In Biochemistry and Genetics of Cellulose Degradation (FEMS Symposium 43) pp. 31 52 Edited by Aubert J.-P., Beguin P., Millet J. New York: Academic Press;
    [Google Scholar]
  39. Xue G. P., Morris R. 1992; Expression of the neuronal surface glycoprotein Thy-1 does not follow appearance of its mRNA in developing mouse Purkinje cells. Journal of Neurochemistry 58 430 440
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/00221287-138-7-1413
Loading
/content/journal/micro/10.1099/00221287-138-7-1413
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error