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. 1990 Apr;56(4):1017–1024. doi: 10.1128/aem.56.4.1017-1024.1990

Cloning, sequence analysis, and expression of genes encoding xylan-degrading enzymes from the thermophile "Caldocellum saccharolyticum".

E Lüthi 1, D R Love 1, J McAnulty 1, C Wallace 1, P A Caughey 1, D Saul 1, P L Bergquist 1
PMCID: PMC184337  PMID: 2111111

Abstract

A lambda recombinant bacteriophage coding for xylanase and beta-xylosidase activity has been isolated from a genomic library of the extremely thermophilic anaerobe "Caldocellum saccharolyticum." Partial Sau3AI fragments of the lambda recombinant DNA were ligated into pBR322. A recombinant plasmid with an insertion of ca. 7 kilobases of thermophilic DNA expressing both enzymatic activities was isolated. The location of the genes has been established by analyzing deletion derivatives, and the DNA sequence of 6.067 kilobases of the insert has been determined. Five open reading frames (ORFs) were found, one of which (ORF1; Mr 40,455) appears to code for a xylanase (XynA) which also acts on o-nitrophenyl-beta-D-xylopyranoside. Another, ORF5 (Mr 56,365), codes for a beta-xylosidase (XynB). The xynA gene product shows significant homology with the xylanases from the alkalophilic Bacillus sp. strain C125 and Clostridium thermocellum.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Bankier A. T., Weston K. M., Barrell B. G. Random cloning and sequencing by the M13/dideoxynucleotide chain termination method. Methods Enzymol. 1987;155:51–93. doi: 10.1016/0076-6879(87)55009-1. [DOI] [PubMed] [Google Scholar]
  2. Bergquist P. L., Love D. R., Croft J. E., Streiff M. B., Daniel R. M., Morgan W. H. Genetics and potential biotechnological applications of thermophilic and extremely thermophilic micro-organisms. Biotechnol Genet Eng Rev. 1987;5:199–244. doi: 10.1080/02648725.1987.10647838. [DOI] [PubMed] [Google Scholar]
  3. Bolivar F., Rodriguez R. L., Greene P. J., Betlach M. C., Heyneker H. L., Boyer H. W., Crosa J. H., Falkow S. Construction and characterization of new cloning vehicles. II. A multipurpose cloning system. Gene. 1977;2(2):95–113. [PubMed] [Google Scholar]
  4. Boucher F., Morosoli R., Durand S. Complete nucleotide sequence of the xylanase gene from the yeast Cryptococcus albidus. Nucleic Acids Res. 1988 Oct 25;16(20):9874–9874. doi: 10.1093/nar/16.20.9874. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Béguin P. Detection of cellulase activity in polyacrylamide gels using Congo red-stained agar replicas. Anal Biochem. 1983 Jun;131(2):333–336. doi: 10.1016/0003-2697(83)90178-1. [DOI] [PubMed] [Google Scholar]
  6. Croft J. E., Love D. R., Bergquist P. L. Expression of leucine genes from an extremely thermophilic bacterium in Escherichia coli. Mol Gen Genet. 1987 Dec;210(3):490–497. doi: 10.1007/BF00327202. [DOI] [PubMed] [Google Scholar]
  7. Gilbert H. J., Sullivan D. A., Jenkins G., Kellett L. E., Minton N. P., Hall J. Molecular cloning of multiple xylanase genes from Pseudomonas fluorescens subsp. cellulosa. J Gen Microbiol. 1988 Dec;134(12):3239–3247. doi: 10.1099/00221287-134-12-3239. [DOI] [PubMed] [Google Scholar]
  8. Grépinet O., Chebrou M. C., Béguin P. Nucleotide sequence and deletion analysis of the xylanase gene (xynZ) of Clostridium thermocellum. J Bacteriol. 1988 Oct;170(10):4582–4588. doi: 10.1128/jb.170.10.4582-4588.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Hall J., Hazlewood G. P., Barker P. J., Gilbert H. J. Conserved reiterated domains in Clostridium thermocellum endoglucanases are not essential for catalytic activity. Gene. 1988 Sep 15;69(1):29–38. doi: 10.1016/0378-1119(88)90375-7. [DOI] [PubMed] [Google Scholar]
  10. Johnson W. C., Moran C. P., Jr, Losick R. Two RNA polymerase sigma factors from Bacillus subtilis discriminate between overlapping promoters for a developmentally regulated gene. Nature. 1983 Apr 28;302(5911):800–804. doi: 10.1038/302800a0. [DOI] [PubMed] [Google Scholar]
  11. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  12. Lee S. F., Forsberg C. W., Rattray J. B. Purification and Characterization of Two Endoxylanases from Clostridium acetobutylicum ATCC 824. Appl Environ Microbiol. 1987 Apr;53(4):644–650. doi: 10.1128/aem.53.4.644-650.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. MacKay R. M., Lo A., Willick G., Zuker M., Baird S., Dove M., Moranelli F., Seligy V. Structure of a Bacillus subtilis endo-beta-1,4-glucanase gene. Nucleic Acids Res. 1986 Nov 25;14(22):9159–9170. doi: 10.1093/nar/14.22.9159. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. McLaughlin J. R., Murray C. L., Rabinowitz J. C. Unique features in the ribosome binding site sequence of the gram-positive Staphylococcus aureus beta-lactamase gene. J Biol Chem. 1981 Nov 10;256(21):11283–11291. [PubMed] [Google Scholar]
  15. Moran C. P., Jr, Lang N., LeGrice S. F., Lee G., Stephens M., Sonenshein A. L., Pero J., Losick R. Nucleotide sequences that signal the initiation of transcription and translation in Bacillus subtilis. Mol Gen Genet. 1982;186(3):339–346. doi: 10.1007/BF00729452. [DOI] [PubMed] [Google Scholar]
  16. Moriyama H., Fukusaki E., Cabrera Crespo J., Shinmyo A., Okada H. Structure and expression of genes coding for xylan-degrading enzymes of Bacillus pumilus. Eur J Biochem. 1987 Aug 3;166(3):539–545. doi: 10.1111/j.1432-1033.1987.tb13547.x. [DOI] [PubMed] [Google Scholar]
  17. Mézes P. S., Yang Y. Q., Hussain M., Lampen J. O. Bacillus cereus 569/H beta-lactamase I: cloning in Escherichia coli and signal sequence determination. FEBS Lett. 1983 Sep 19;161(2):195–200. doi: 10.1016/0014-5793(83)81006-0. [DOI] [PubMed] [Google Scholar]
  18. Nielsen J. B., Lampen J. O. Membrane-bound penicillinases in Gram-positive bacteria. J Biol Chem. 1982 Apr 25;257(8):4490–4495. [PubMed] [Google Scholar]
  19. O'Neill G., Goh S. H., Warren R. A., Kilburn D. G., Miller R. C., Jr Structure of the gene encoding the exoglucanase of Cellulomonas fimi. Gene. 1986;44(2-3):325–330. doi: 10.1016/0378-1119(86)90197-6. [DOI] [PubMed] [Google Scholar]
  20. Paul J. R., Trask J. D. Occurrence and Recovery of the Virus of Infantile Paralysis from Sewage. Am J Public Health Nations Health. 1942 Mar;32(3):235–239. doi: 10.2105/ajph.32.3.235. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Saul D. J., Williams L. C., Love D. R., Chamley L. W., Bergquist P. L. Nucleotide sequence of a gene from Caldocellum saccharolyticum encoding for exocellulase and endocellulase activity. Nucleic Acids Res. 1989 Jan 11;17(1):439–439. doi: 10.1093/nar/17.1.439. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Short J. M., Fernandez J. M., Sorge J. A., Huse W. D. Lambda ZAP: a bacteriophage lambda expression vector with in vivo excision properties. Nucleic Acids Res. 1988 Aug 11;16(15):7583–7600. doi: 10.1093/nar/16.15.7583. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Sipat A., Taylor K. A., Lo R. Y., Forsberg C. W., Krell P. J. Molecular cloning of a xylanase gene from Bacteroides succinogenes and its expression in Escherichia coli. Appl Environ Microbiol. 1987 Mar;53(3):477–481. doi: 10.1128/aem.53.3.477-481.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Stalker D. M., McBride K. E., Malyj L. D. Herbicide resistance in transgenic plants expressing a bacterial detoxification gene. Science. 1988 Oct 21;242(4877):419–423. doi: 10.1126/science.242.4877.419. [DOI] [PubMed] [Google Scholar]
  26. Teather R. M., Wood P. J. Use of Congo red-polysaccharide interactions in enumeration and characterization of cellulolytic bacteria from the bovine rumen. Appl Environ Microbiol. 1982 Apr;43(4):777–780. doi: 10.1128/aem.43.4.777-780.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Wain-Hobson S., Sonigo P., Danos O., Cole S., Alizon M. Nucleotide sequence of the AIDS virus, LAV. Cell. 1985 Jan;40(1):9–17. doi: 10.1016/0092-8674(85)90303-4. [DOI] [PubMed] [Google Scholar]
  28. Watson M. E. Compilation of published signal sequences. Nucleic Acids Res. 1984 Jul 11;12(13):5145–5164. doi: 10.1093/nar/12.13.5145. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. West C. A., Elzanowski A., Yeh L. S., Barker W. C. Homologues of catalytic domains of Cellulomonas glucanases found in fungal and Bacillus glycosidases. FEMS Microbiol Lett. 1989 May;50(1-2):167–172. doi: 10.1016/0378-1097(89)90479-5. [DOI] [PubMed] [Google Scholar]
  30. Wong K. K., Tan L. U., Saddler J. N. Multiplicity of beta-1,4-xylanase in microorganisms: functions and applications. Microbiol Rev. 1988 Sep;52(3):305–317. doi: 10.1128/mr.52.3.305-317.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Yang R. C., MacKenzie C. R., Narang S. A. Nucleotide sequence of a Bacillus circulans xylanase gene. Nucleic Acids Res. 1988 Jul 25;16(14B):7187–7187. doi: 10.1093/nar/16.14.7187. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Yanisch-Perron C., Vieira J., Messing J. Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. Gene. 1985;33(1):103–119. doi: 10.1016/0378-1119(85)90120-9. [DOI] [PubMed] [Google Scholar]

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