1887

Abstract

subsp. was shown to express extracellular xylanases. Genes encoding these enzymes were isolated from a gene library of subsp. DNA, constructed in bacteriophage λ47.1. One of the phages (PXC) that expressed xylanase also conferred the ability to hydrolyse carboxymethylcellulose. An 11·8 kb III DNA restriction fragment and a 6·2 kb RI DNA fragment were subcloned from two distinct xylanase-expressing phages, into pUC18, to yield recombinant plasmids pGHJ4 and pGHJ5 respectively. Cells of harbouring either of these two plasmids, or plasmid pJHH1 (comprising the cellulase gene from PXC, previously cloned on a 7·3 kb partial RI DNA fragment in pUC18), expressed xylanase activity. The positions of the xylanase genes in the recombinant plasmids were elucidated by subcloning and transposon mutagenesis. In pJHH1 the xylanase gene was adjacent to the DNA region encoding the endoglucanase. The polysaccharide-degrading genes in pJHH1 were transcribed from different promotors. Hybridization studies revealed that the xylanase genes encoded by pGHJ4 and pGHJ5 showed strong homology. All three cloned enzymes cleaved -nitrophenyl --glucopyranoside and 4-methylumbelliferyl --cellobioside. Xylan and glucose did not affect expression of xylanase in strains harbouring pJHH1, pGHJ4 or pGHJ5.

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/content/journal/micro/10.1099/00221287-134-12-3239
1988-12-01
2021-10-25
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References

  1. Bagdasarian M., Lurz R., Ruckert B., Franklin F.C.H., Bagdasarian M.M., Frey J., Timmis K.N. 1981; Specific-purpose plasmid cloning vectors. II. Broad host range, high copy number, RSF 1010-derived vectors and a host-vector system for gene cloning in Pseudomonas.. Gene 16:237–247
    [Google Scholar]
  2. Bernier R. Jr Drigeuz H., Desrochers M. 1983; Molecular cloning of a Bacillus subtilisxylanase gene in Escherichia coli.. Gene 26:59–65
    [Google Scholar]
  3. Biely P. 1985; Microbial xylanolytic systems.. Trends in Biotechnology 3:286–290
    [Google Scholar]
  4. Biely P., Mislovicova D., Toman R. 1985; Soluble chromogenic substrates for the assay of endo-l,4-xylanases and endo-l,4-glucanases.. Analytical Biochemistry 144:142–146
    [Google Scholar]
  5. Clewell D.B., Helinski D.R. 1969; Supercoiled circular DNA-protein complex in E. coli. Purification and induced conversion to an open circular form.. Proceedings of the National Academy of Sciences of the United States of America 62:1159–1166
    [Google Scholar]
  6. Cohen S.N., Chang A.C.Y., Hsu L. 1972; Nonchromosomal antibiotic resistance in bacteria: genetic transformation of E. coli by R-factor DNA.. Proceedings of the National Academy of Sciences of the United States of America 69:2110–2114
    [Google Scholar]
  7. Gilbert H.J., Blazek R., Bullman H.M.S., Minton N.P. 1986; Cloning and expression of the Erwinia chrysanthemi asparaginase gene in Escherichia coli and Erwinia carotovora.. Journal of General Microbiology 132:151–160
    [Google Scholar]
  8. Gilbert H.J., Jenkins G., Sullivan D.A., Hall J. 1987; Evidence for multiple carboxymethylcellu- lase genes in Pseudomonas fluorescens subsp. cellulosa.. Molecular and General Genetics 210:551–556
    [Google Scholar]
  9. Guyer M.S. 1978; The sequence of F is an insertion sequence.. Journal of Molecular Biology 126:347–365
    [Google Scholar]
  10. Hall J., Gilbert H.J. 1988; The nucleotide sequence of a carboxymethylcellulase gene from Pseudomonas fluorescens subsp. cellulosa.. Molecular and General Genetics 213:112–117
    [Google Scholar]
  11. Holmes D.S., Quigley M. 1981; A rapid boiling method for the preparation of bacterial plasmids.. Analytical Biochemistry 114:193–197
    [Google Scholar]
  12. Honda H., Kudo T., Horikoshi K. 1985; Molecular cloning and expression of the xylanase gene of alkalophilic Bacillus sp. strain C-125 in Escherichia coli.. Journal of Bacteriology 163:784–785
    [Google Scholar]
  13. Lee S.F., Forsberg C.W., Raittray J.B. 1987; Purification and characterization of two endoxylan- ases from Clostridium acetobutylicum ATCC 8824.. Applied and Environmental Microbiology 53:644–650
    [Google Scholar]
  14. Maniatis T., Fritsch E., Sambrook J. 1982 Molecular Cloning: a Laboratory Manual. Cold Spring Harbor, NY:: Cold Spring Harbor Laboratory.;
    [Google Scholar]
  15. Meyers J.A., Sanchez D., Elwell L.P., Falkow S. 1976; Simple agarose gel electrophoresis method for the identification and characterisation of plasmid deoxyribonucleic acid.. Journal of Bacteriology 127:1529–1538
    [Google Scholar]
  16. Millet J., Petre D., Beguin P., Raynaud O., Aubert J.-P. 1985; Cloning of ten distinct DNA fragments of Clostridium thermocellum coding for cellulases.. FEMS Microbiology Letters 29:145–149
    [Google Scholar]
  17. Minton N.P. 1984; Improved plasmid vectors for the isolation of translational lac gene fusions.. Gene 31:269–273
    [Google Scholar]
  18. Minton N.P., Atkinson T., Sherwood R.F. 1983; Molecular cloning of Pseudomonas car boxy- peptidase G2 gene and its expression in Escherichia coli and Pseudomonas putida.. Journal of Bacteriology 156:1222–1227
    [Google Scholar]
  19. Norrander J., Kemp T., Messing J. 1983; Construction of improved M13 vectors using oligo- deoxynucleotide-directed mutagenesis.. Gene 26:101–116
    [Google Scholar]
  20. Panbangred W., Kondo T., Negoro S., Shinmyo A., Okada H. 1983; Molecular cloning of the genes for xylan degradation of Bacillus pumilus and their expression in Escherichia coli.. Molecular and General Genetics 192:335–341
    [Google Scholar]
  21. Rigby P.W.J., Dieckmann M., Rhodes C., Berg P. 1977; Labelling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I.. Journal of Molecular Biology 113:237–251
    [Google Scholar]
  22. 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]
  23. Taylor K.A., Crosby B., Mcgavin M., Forsberg C.W., Thomas D.Y. 1987; Characteristics of the endoglucanase encoded by a cel gene from Bacteroides succinogenes expressed in Escherichia coli.. Applied and Environmental Microbiology 53:41–46
    [Google Scholar]
  24. Van Tilbeurgh H., Claeyssens M., De Bruyne C.K. 1982; The use of 4-methylumbelliferyl and other chromophoric glycosides in the study of cellulolytic enzymes.. FEBS Letters 149:152–156
    [Google Scholar]
  25. Yamane K., Suzuki H., Hirotani M., Ozawa H., Nisizawa K. 1970; Effect of nature and supply of carbon sources on cellulase formation in Psetuiomonas fluorescens var. cellulosa.. Journal of Biochemistry 67:9–28
    [Google Scholar]
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