1887

Microbiology Society logo

Volume 139, Issue 2

Gene sequence and properties of Cell, a family E endoglucanase from Free

Abstract

The gene, coding for endoglucanase I (Cell), consists of an open reading frame (ORF) of 2640 nucleotides and codes for a protein of 98531. The ORF was confirmed as by comparing the N-terminal sequence of purified recombinant Cell with that deduced from the nucleotide sequence. Cell hydrolysed lichenan and carboxymethylcellulose, but was principally active against barley-glucan. It exhibited significant sequence identity with subfamily E endoglucanases, and by analogy with others in this group contains a catalytic domain of around 500 residues located in the N-terminal half of the protein. The C-terminal region of Cell was highly homologous with the cellulose-binding domain of the non-catalytic cellulosome subunit, S1. A repeated segment, previously shown to be highly conserved in xylanase Z and in other endoglucanases from , was absent from Cell. Antiserum raised against purified recombinant Cell cross-reacted with proteins contained in the cellulosomes of two strains of , suggesting that Cell is either a component of the cellulosome or is homologous to other cellulosome proteins. A second gene, located upstream of , consisted of an ORF of 1671 nucleotides, coding for a protein of 61042. Based on its homology with the tar gene product, the polypeptide encoded by the second gene is tentatively identified as a sensory transducer.

  • Received:
  • Accepted:
  • Published Online:
© Society for General Microbiology 1993
Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-139-2-307
1993-02-01
2024-04-25
Loading full text...

Full text loading...

/deliver/fulltext/micro/139/2/mic-139-2-307.html?itemId=/content/journal/micro/10.1099/00221287-139-2-307&mimeType=html&fmt=ahah

References

  1. Béguin P. 1990; Molecular biology of cellulose degradation. Annual Review of Microbiology 44:219–248
     [Google Scholar]
  2. Béguin P., Cornet P., Aubert J.P. 1985; Sequence of a cellulase gene of the thermophilic bacterium Clostridium thermocellum. Journal of Bacteriology 162:102–105
     [Google Scholar]
  3. Berger E., Jones W.A., Jones D.T., Woods D.R. 1990; Sequencing and expression of a cellodextrinase(cedl)gene from Butyrivibrio fibrisolvens HI7c cloned in Escherichia coli. Molecular and General Genetics 223:310–318
     [Google Scholar]
  4. Birnboim H.C., Doly J. 1979; A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Research 7:1513–1523
     [Google Scholar]
  5. Chauvaux S., Béguin P., Aubert J.P., Bhat K.M., Gow L.A., Wood T.M., Bairoch A. 1990; Calcium-binding affinity and calcium-enhanced activity of Clostridium thermocellum endoglucanase D. Biochemical Journal 265:261–265
     [Google Scholar]
  6. Coutinho J.B., Moser B., Kilburn D.G., Warren R.A.J., Miller R.C. 1991; Nucleotide sequence of the endoglucanase C gene (cenC) of Cellulomonas fimi, its high-level expression in Escherichia coli, and characterization of its products. Molecular Microbiology 5:1221–1233
     [Google Scholar]
  7. Durrant A.J., Hall J., Hazlewood G.P., Gilbert H.J. 1991; The non-catalytic C-terminal region of endoglucanase E from Clostridium thermocellum contains a cellulose-binding domain. Biochemical Journal 273:289–293
     [Google Scholar]
  8. Gardina P., Conway C., Kossman M., Manson M. 1992; Aspartate and maltose-binding protein interact with adjacent sites in the tar chemitactic signal transducer of Escherichia coli. Journal of Bacteriology 17:1528–1536
     [Google Scholar]
  9. Giorda R., Ohmachi T., Shaw D.R., Ennis H.L. 1990; A shared internal threonine-glutamic acid-threonine-proline repeat defines a family of Dictyostelium discoideum spore germination proteins. Biochemistry 29:7264–7269
     [Google Scholar]
  10. Grabnitz F.-R., Rücknagel K.P., Seib M., Staudenbauer W.L. 1989; Nucleotide sequence of the Clostridium thermocellum bglB gene encoding thermostable β-glucosidase B : homology to fungal β-glucosidases. Molecular and General Genetics 217:70–76
     [Google Scholar]
  11. Grépinet O., Béguin P. 1986; Sequence of the cellulase gene of Clostridium thermocellum coding for endoglucanase B. Nucleic Acids Research 14:1791–1799
     [Google Scholar]
  12. Grépinet O., Chebrou M.C., Béguin P. 1988; Nucleotide sequence and deletion analysis of the xylanase gene (xynZ) of Clostridium thermocellum. Journal of Bacteriology 179:4582–4588
     [Google Scholar]
  13. Hall J., Gilbert H.J. 1988; The nucleotide sequence of a carboxymethylcellulase from Pseudomonas fluorescens subsp.cellulosa. Molecular and General Genetics 213:112–117
     [Google Scholar]
  14. Hall J., Hazlewood G.P., Barker P.J., Gilbert H.J. 1988; Conserved reiterated domains in Clostridium thermocellum endoglucanases are not essential for catalytic activity. Gene 69:29–38
     [Google Scholar]
  15. 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, β-glucosidase and xylanase genes cloned in Escherichia coli. FEMS Microbiology Letters 51:231–236
     [Google Scholar]
  16. Hazlewood G.P., Davidson K., Clarke J.H., Durrant A.J., Hall J., Gilbert H.J. 1990a; Endoglucanase E produced at high level in Escherichia coli as a lacZ’ fusion protein, is part of the Clostridium thermocellum cellulosome. Enzyme and Microbial Technology 12:656–662
     [Google Scholar]
  17. Hazlewood G.P., Davidson K., Laurie J.I., Romaniec M.P.M., Gilbert H.J. 1990b; Cloning and sequencing of the celA gene encoding endoglucanase A of Butyrivibio fibrisolvens strain A46. Journal of General Microbiology 136:2089–2097
     [Google Scholar]
  18. Henrissat B., Claeyssens M., Tomme P., Lemesle L., Mornon J.P. 1989; Cellulase families revealed by hydrophobic cluster analysis. Gene 81:83–95
     [Google Scholar]
  19. Hunkapillar M.W., Hewick R.E., Dreyer W.J., Hood L.E. 1983; High sensitivity sequencing with gas-phase sequenator. Methods in Enzymology 91:399–413
     [Google Scholar]
  20. Jauris S., Rücknagel K.P., Schwarz W.H., Kratzsch P., Bronnenmeier K., Staudenbauer W.L. 1990; Sequence analysis of the Clostridium stercorarium celZ gene encoding a thermoactive cellulase (Avicelase I): identification of catalytic and cellulose-binding domains. Molecular and General Genetics 223:258–267
     [Google Scholar]
  21. Joliff G., Béguin P., Aubert J.P. 1986; Nucleotide sequence of the cellulase gene celD encoding endoglucanase D of Clostridium thermocellum. Nucleic Acids Research 14:8605–8613
     [Google Scholar]
  22. Kohring S., Wiegel J., Mayer F. 1990; Subunit composition and glycosidic activities of the cellulase complex from Clostridium thermocellum JW20. Applied and Environmental Microbiology 56:3798–3804
     [Google Scholar]
  23. Krikos A., Mutoh N., Boyd A., Simon M.I. 1983; Sensory transducers of E. coli are composed of discrete structural and functional domains. Cell 33:615–622
     [Google Scholar]
  24. Laemmli U.K. 1970; Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature; London: 227680–685
     [Google Scholar]
  25. Lamed R., Bayer E.A. 1988; The cellulosome of Clostridium thermocellum. Advances in Applied Microbiology 33:1–46
     [Google Scholar]
  26. Mayer F., Coughlan M.P., Mori Y., Ljungdahl L.G. 1987; Macromolecular organization of the cellulolytic enzyme complex of Clostridium thermocellum as revealed by electron microscopy. Applied and Environmental Microbiology 53:2785–2792
     [Google Scholar]
  27. McLaughlin J.R., Murray C.L., Rabinowitz J.C. 1981; Unique features in the ribosome binding site sequence of the gram positive Staphylococcus aureusβ-lactamase gene. Journal of Biological Chemistry 256:11283–11291
     [Google Scholar]
  28. Meinke A., Braun C., Gilkes N.R., Kilburn D.G., Miller R.C., Warren R.A.J. 1991; Unusual sequence organization in CenB, an inverting endoglucanaseCellulomonasfimi. Journal of Bacteriology 173:308–314
     [Google Scholar]
  29. Morag E., Bayer E.A., Lamed R. 1990; Relationship of cellulosomal and noncellulosomal xylanases of Clostridium thermocellum to cellulose-degrading enzymes. Journal of Bacteriology 172:6098–6105
     [Google Scholar]
  30. Morag E., Halevy A.L., Bayer E., Lamed R. 1991; Isolation and properties of a major cellobiohydrolase from the cellulosome of Clostridium thermocellum. Journal of Bacteriology 173:4155–4162
     [Google Scholar]
  31. Morag E., Bayer E.A., Lamed R. 1992; Affinity digestion for the near-total recovery of purified cellulosome from Clostridium thermocellum. Enzyme and Microbial Technology 14:289–292
     [Google Scholar]
  32. Murphy G., Kavanagh T. 1988; Speeding-up the sequencing of double-stranded DNA. Nucleic Acids Research 16:5198
     [Google Scholar]
  33. Navarro A., Chebrou M.C., Béguin P., Aubert J.P. 1991; Nucleotide sequence of the cellulase gene celF of Clostridium thermocellum. Research in Microbiology 142:927–936
     [Google Scholar]
  34. Norrander J., Kemp T., Messing J. 1983; Construction of improved Ml3 vectors using oligodeoxynucleotide-directed mutagenesis. Gene 26:101–116
     [Google Scholar]
  35. Poole D.M., Morag E., Lamed R., Bayer E.A., Hazlewood G.P., Gilbert H.J. 1992; Identification of the cellulose binding domain of cellulosome subunit SI from Clostridium thermocellum YS. FEMS Microbiology Letters in the Press
    [Google Scholar]
  36. Romaniec M.P.M., Clarke N.G., Hazlewood G.P. 1987; Molecular cloning of Clostridium thermocellum DNA and the expression of further novel endo-β-1,4-glucanase genes in Escherichia coli. Journal of General Microbiology 133:1297–1307
     [Google Scholar]
  37. Sambrook J., Fritsch E.F., Maniatis T. 1989 Molecular Cloning: a Laboratory Manual, 2nd edn.. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
     [Google Scholar]
  38. Sanger F., Nicklen S., Coulson A.R. 1977; DNA sequencing with chain-terminating inhibitors. Proceedings of the National Academy of Sciences of the United States of America 74:5463
     [Google Scholar]
  39. Schwarz W.H., Schimming S., Rücknagel K.P., Burgschwaiger S., Kreil G., Staudenbauer W.L. 1988; Nucleotide sequence of the celC gene encoding endoglucanase C of Clostridium thermocellum. Gene 63:23–30
     [Google Scholar]
  40. Sedmak J.J., Grossberg S.E. 1977; A rapid, sensitive and versatile assay for protein using Coomassie Brilliant Blue G250. Analytical Biochemistry 69:544–552
     [Google Scholar]
  41. Tokatlidis K., Salamitou S., Béguin P., Dhurjati P., Aubert J.P. 1991; Interaction of the duplicated segment carried by Clostridium thermocellum cellulases with cellulosome components. FEBS Letters 291:185–188
     [Google Scholar]
  42. Tomme P., Chauvaux S., Béguin P., Millet J., Aubert J.P., Claeyssens M. 1991; Identification of a histidyl residue in the active center of endoglucanase D from Clostridium thermocellum. Journal of Biological Chemistry 266:10313–10318
     [Google Scholar]
  43. Towbin H., Staehelin T., Gordon J. 1979; Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets : procedure and some applications. Proceedings of the National Academy of Sciences of the United States of America 76:4350–4354
     [Google Scholar]
  44. Tucker M.L., Durbin M.L., Clegg M.T., Lewis L.N. 1987; Avocado cellulase: nucleotide sequence of a putative full-length cDNA clone and evidence for a small gene family. Plant Molecular Biology 9:197–203
     [Google Scholar]
  45. Von Heijne G. 1988; Transcending the impenetrable: how proteins come to terms with membranes. Biochimica et Biophysica Acta 947:307–333
     [Google Scholar]
  46. Yagüe E.E., Béguin P., Aubert J.P. 1990; Nucleotide sequence and deletion analysis of the cellulase-encoding gene celH of Clostridium thermocellum. Gene 89:61–67
     [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/00221287-139-2-307
Loading
Gene sequence and properties of Cell, a family E endoglucanase from Clostridium thermocellum
Microbiology 139, 307 (1993); https://doi.org/10.1099/00221287-139-2-307
/content/journal/micro/10.1099/00221287-139-2-307
/content/journal/micro/10.1099/00221287-139-2-307
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