1887

Microbiology Society logo

Volume 137, Issue 1

Expression of two xylanase genes from the rumen cellulolytic bacterium 17 cloned in pUC13 Free

Abstract

Two distinct xylanase genes (designated and ) were subcloned in pUC13 from non-homologous restriction fragments of 17 DNA originally isolated in EMBL3. The products of the two genes showed similar pH optima for hydrolysis of oat spelt xylan (around 5·5) and had little or no activity against carboxymethylcellulose. Trace activities against -nitrophenyl (pNP) cellobioside and pNP-xyloside were detected in clones containing , but not in one harbouring . The xylanase associated with clones carrying produced mainly xylobiose and xylose from xylan and did not give hydrolysis of xylobiose, while that encoded by produced mainly xylobiose and higher xylo-oligosaccharides from xylan. There was evidence of increased expression, at the RNA level, of these two genes, and of another cloned region encoding multiple activities including xylanase, in 17 grown with xylan, as compared with cellobiose, as energy source. Total cell-associated xylanase and -xylosidase activities, and supernatant xylanase activity, were shown to be similarly induced in xylan-grown , 17.

  • Received:
  • Accepted:
  • Revised:
  • Published Online:
© Society for General Microbiology, 1991
Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-137-1-123
1991-01-01
2024-04-23
Loading full text...

Full text loading...

/deliver/fulltext/micro/137/1/mic-137-1-123.html?itemId=/content/journal/micro/10.1099/00221287-137-1-123&mimeType=html&fmt=ahah

References

  1. Barros M. E. C., Thomson J. A. 1987; Cloning and expression in Escherichia coli of a cellulase gene from Ruminococcus flavefaciens . Journal of Bacteriology 169:1760–1762
     [Google Scholar]
  2. Béguin P., Millet J., Grépinet O., Navarro A., Amit A., Poljak R., Aubert J-P. 1988; The cell (cellulose degradation) genes of Clostridium thermocellum . In Biochemistry and Genetics of Cellulose Degradation267–282 Aubert P., Beguin P., Millet J. New York & London: Academic Press;
     [Google Scholar]
  3. Biely P., Mislovicova D., Toman R. 1988; Remazol Brilliant Blue-xylan : a soluble chromogenic substrate for xylanases. Methods in Enzymology 160A536–541
     [Google Scholar]
  4. Bryant Μ. P. 1986; Genus Ruminococcus Sijpesteijn 1948. In Bergey’s Manual of Systematic Bacteriology 21093–1097 Sneath P. H. A., Moir M. S., Sharpe M. E., Holt J. G. Baltimore: Williams & Wilkins;
     [Google Scholar]
  5. Bryant M. P. 1972; Commentary on the Hungate technique for culture of anaerobic bacteria. American Journal of Clinical Nutrition 25:1324–1328
     [Google Scholar]
  6. Chirgwin J. M., Przybala A. E., Macdonald R. J., Rutter W. J. 1979; Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease. Biochemistry 18:5294–5299
     [Google Scholar]
  7. Dehority B. A., Scott H. W. 1967; Extent of cellulose and hemicellulose digestion in various forages by pure cultures of rumen bacteria. Journal of Dairy Science 50:1136–1141
     [Google Scholar]
  8. Dower W. J., Miller J. F., Ragsdale C. W. 1988; High efficiency transformation of Escherichia coli by high voltage electroporation. Nucleic Acids Research 16:6127–6145
     [Google Scholar]
  9. Flint H. J., Wilkening J. 1986; Cloning of the arg12 gene of Neurospora crassa and regulation of its transcript by cross-pathway amino acid control. Molecular and General Genetics 203:110–116
     [Google Scholar]
  10. Flint H. J., McPherson C. A., Bisset J. 1989; Molecular cloning of genes from Ruminococcus flavefaciens encoding xylanase and β(1,3-1,4)-glucanase activities. Applied and Environmental Microbiology 55:1230–1233
     [Google Scholar]
  11. Fontana J. D., Gedara M., Blumel M., Schneider H., Mackenzie C. R., Johnson K. G. 1988; α4-OMethyl-dglucuronidase component of xylanolytic complexes. Methods in Enzymology 160A:560–571
     [Google Scholar]
  12. Gilbert H. J., Sullivan D. A., Jenkins G., Kellet L. E., Minton N. P., Hall J. P. 1988; Molecular cloning of multiple xylanase genes from Pseudomonas fluorescens subsp. cellulosa . Journal of General Microbiology 134:3239–3247
     [Google Scholar]
  13. 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]
  14. Grépinet O., Chebrou M.-C., Béguin J-P. 1988; Nucleotide sequence and deletion analysis of the xylanase gene (xynZ) of Clostridium thermocellum . Journal of Bacteriology 170:4582–4588
     [Google Scholar]
  15. Greve L. C, Labavitch J. M., Stack R. J., Hungate R. E. 1984; Muralytic activities of Ruminococcus albus 8. Applied and Environmental Microbiology 47:1141–1145
     [Google Scholar]
  16. Halliwell G., Bryant M. P. 1963; The cellulolytic activity of pure strains of bacteria from the rumen of cattle. Journal of General Microbiology 32:441–448
     [Google Scholar]
  17. Hazlewood G. P., Romaniec M. P. M., Davidson K., Grépinet O., Béguin P., Millet J., Raynaud O., Aubert J.-P. 1988; A catalogue of Clostridium thermocellum endoglucanase, βglucosidase and xylanase genes cloned in Escherichia coli . FEMS Microbiology Letters 51:231–236
     [Google Scholar]
  18. Howard G. T., White B. A. 1988; Molecular cloning and expression of cellulase genes from Ruminococcus albus and in Escherichia coli bacteriophage. Applied and Environmental Microbiology 54:1752–1755
     [Google Scholar]
  19. Huang C.-M., Kelly W. J., Asmundson R. V., Yu P.-K. 1989; Molecular cloning and expression of multiple cellulase genes of Ruminococcus flavefaciens strain 186 in Escherichia coli . Applied Microbiology and Biotechnology 31:265–271
     [Google Scholar]
  20. Hungate R. E., Stack R. J. 1982; Phenylpropanoic acid: growth factor for Ruminococcus albus . Applied and Environmental Microbiology 44:79–83
     [Google Scholar]
  21. Laemmli U. K. 1970; Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature, London 227:680–685
     [Google Scholar]
  22. Lever M. 1977; Carbohydrate determination with 4-hydroxybenzoic acid hydrazide (PAHBAH): effect of bismuth on the reaction. Analytical Biochemistry 81:21–27
     [Google Scholar]
  23. Lüthi E., Love D. R., McNulty J., Wallace C, Caughey P. A., Saul D., Bergquist P. L. 1990; Cloning, sequence analysis and expression of genes encoding xylan degrading enzymes from the thermophile ‘Caldocellum saccharolyticum’. Applied and Environmental Microbiology 56:1017–1024
     [Google Scholar]
  24. Maniatis T., Fritsch E. F., Sambrook J. 1982 Molecular Cloning - a Laboratory Manual Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
     [Google Scholar]
  25. Mannarelli B. M., Evans S., Lee D. 1990; Cloning, sequencing and expression of a xylanase gene from the anaerobic ruminal bacterium Butyrivibrio fibrisolvens . Journal of Bacteriology 172:4247–4254
     [Google Scholar]
  26. Morris E. J., Cole O. J. 1987; Relationship between cellulolytic activity and adhesion to cellulose in Ruminococcus albus . Journal of General Microbiology 133:1023–1032
     [Google Scholar]
  27. Ohmiya K., Nagashma K., Kajino T., Goto E., Tsukada A., Shimizu S. 1988; Cloning of the cellulase gene from Ruminococcus albus and its expression in E. coli . Applied and Environmental Microbiology 54:1511–1515
     [Google Scholar]
  28. Pettipher G. L., Latham M. J. 1979a; Characteristics of enzymes produced by Ruminococcus flavefaciens which degrade plant cell walls. Journal of General Microbiology 110:21–27
     [Google Scholar]
  29. Pettipher G. L., Latham M. J. 1979b; Production of enzymes degrading plant cell walls and fermentation of cellobiose by Ruminococcus flavefaciens in batch and continuous culture. Journal of General Microbiology 110:29–38
     [Google Scholar]
  30. Romaniec M. P. M., Davidson K., White B. A., Hazlewood G. P. 1989; Cloning of Ruminococcus albus endo-β1,4-glucanase and xylanase genes. Letters in Applied Microbiology 9:101–104
     [Google Scholar]
  31. Sewell G. W., Utt E. A., Hespell R. B., Mackenzie K. F., Ingram L. O. 1989; Identification of the Butyrivibrio fibrisolvens xylosidase gene (xylB) coding region and its expression in Escherichia coli . Applied and Environmental Microbiology 55:306–311
     [Google Scholar]
  32. Sipat A., Taylor K. A., Lo R. Y. C, Forsberg C. W., Krell P. J. 1987; Molecular cloning of a xylanase gene from Bacteroides succinogenes and its expression in Escherichia coli . Applied and Environmental Microbiology 53:477–481
     [Google Scholar]
  33. Somogyi M. 1952; Notes on sugar determination. Journal of Biological Chemistry 195:19–23
     [Google Scholar]
  34. 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]
  35. Ware G. E., Bauchop T., Gregg K. 1989; The isolation and comparison of cellulase genes from two strains of Ruminococcus albus . Journal of General Microbiology 135:921–930
     [Google Scholar]
  36. West C. A., Elzanowski A., Yeh L. S., Barber W. C. 1989; Homologues of catalytic domains of Cellulomonas glucanases found in fungal and Bacillus glycosidases. FEMS Microbiology Letters 59:167–172
     [Google Scholar]
  37. Whitehead T. R., Hespell R. B. 1989; Cloning and expression of a xylanase gene from Bacteroides ruminicola 23. Applied and Environmental Microbiology 55:893–896
     [Google Scholar]
  38. Whitehead T. R., Hespell R. B. 1990; Heterologous expression of the Bacteroides ruminocola xylanase gene in Bacteroides fragilis and Bacteroides uniformis . FEMS Microbiology Letters 66:61–66
     [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/00221287-137-1-123
Loading
Expression of two xylanase genes from the rumen cellulolytic bacterium Ruminococcus flavefaciens 17 cloned in pUC13
Microbiology 137, 123 (1991); https://doi.org/10.1099/00221287-137-1-123
/content/journal/micro/10.1099/00221287-137-1-123
/content/journal/micro/10.1099/00221287-137-1-123
Loading

Data & Media loading...

Most cited Most Cited RSS feed

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