Skip to main content
NCBI home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1986 Dec;168(3):1272–1276. doi: 10.1128/jb.168.3.1272-1276.1986

Cloning and expression in Escherichia coli of the gene for an Arthrobacter beta-(1----3)-glucanase.

K Doi, A Doi
PMCID: PMC213633  PMID: 3096974

Abstract

When inserted in the correct orientation at the BamHI site of plasmid YRp7, an 8.6-kilobase BamHI fragment of Arthrobacter sp. strain YCWD3 DNA gave Escherichia coli HB101 cells harboring the recombinant plasmid pBX20 the ability to lyse bakers' yeast cell walls or bakers' yeast glucan in agar medium. An extract of the transformed E. coli cells contained an endo-beta-(1----3)-glucanase with the same activity pattern as that of glucanase I produced by Arthrobacter sp. strain YCWD3. Although part of the glucanase activity was contributed by apparently defective molecules, two protein species were found which had high lytic activity on yeast cell walls and adsorbed to microcrystalline cellulose, and both had a single constituent polypeptide with a molecular weight of about 55,000, as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. In these properties the protein species were indistinguishable from those glucanase I protein species of Arthrobacter sp. strain YCWD3 which we believe are nearly the intact molecule. We conclude that the cloned fragment of Arthrobacter sp. strain YCWD3 DNA contains the structural gene for glucanase I. A recombinant plasmid obtained by subcloning a PstI fragment of pBX20 into pBR322 caused the transformed E. coli cells to produce apparently defective glucanase molecules only. This observation serves as additional supporting evidence for our conclusion.

Full text

PDF
1272

Images in this article

1273Image
on p.1273
1275Image
on p.1275

Selected References

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

  1. Bacon J. S., Gordon A. H., Jones D., Taylor I. F., Webley D. M. The separation of beta-glucanases produced by Cytophaga johnsonii and their role in the lysis of yeast cell walls. Biochem J. 1970 Nov;120(1):67–78. doi: 10.1042/bj1200067. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. CHESTERS C. G., BULL A. T. The enzymic degradation of laminarin. 2. The multicomponent nature of fungal laminarinases. Biochem J. 1963 Jan;86:31–38. doi: 10.1042/bj0860031. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Clarke L., Carbon J. Functional expression of cloned yeast DNA in Escherichia coli: specific complementation of argininosuccinate lyase (argH) mutations. J Mol Biol. 1978 Apr 25;120(4):517–532. doi: 10.1016/0022-2836(78)90351-0. [DOI] [PubMed] [Google Scholar]
  4. DAVIS B. J. DISC ELECTROPHORESIS. II. METHOD AND APPLICATION TO HUMAN SERUM PROTEINS. Ann N Y Acad Sci. 1964 Dec 28;121:404–427. doi: 10.1111/j.1749-6632.1964.tb14213.x. [DOI] [PubMed] [Google Scholar]
  5. Doi K., Doi A., Fukui T. Joint action of two glucanases produced by Arthrobacter in spheroplast formation from baker's yeast. J Biochem. 1971 Oct;70(4):711–714. doi: 10.1093/oxfordjournals.jbchem.a129686. [DOI] [PubMed] [Google Scholar]
  6. Doi K., Doi A., Ozaki T., Fukui T. Heterogeneity of the lytic activity for yeast cell wall observed among the components of an Arthrobacter glucanase. J Biochem. 1973 Mar;73(3):667–670. doi: 10.1093/oxfordjournals.jbchem.a130127. [DOI] [PubMed] [Google Scholar]
  7. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  8. 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]
  9. Mayer H., Grosschedl R., Schütte H., Hobom G. ClaI. a new restriction endonuclease from Caryophanon latum L. Nucleic Acids Res. 1981 Oct 10;9(19):4833–4845. doi: 10.1093/nar/9.19.4833. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Nossal N. G., Heppel L. A. The release of enzymes by osmotic shock from Escherichia coli in exponential phase. J Biol Chem. 1966 Jul 10;241(13):3055–3062. [PubMed] [Google Scholar]
  11. Struhl K., Stinchcomb D. T., Scherer S., Davis R. W. High-frequency transformation of yeast: autonomous replication of hybrid DNA molecules. Proc Natl Acad Sci U S A. 1979 Mar;76(3):1035–1039. doi: 10.1073/pnas.76.3.1035. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Sutcliffe J. G. Complete nucleotide sequence of the Escherichia coli plasmid pBR322. Cold Spring Harb Symp Quant Biol. 1979;43(Pt 1):77–90. doi: 10.1101/sqb.1979.043.01.013. [DOI] [PubMed] [Google Scholar]
  13. Tschumper G., Carbon J. Sequence of a yeast DNA fragment containing a chromosomal replicator and the TRP1 gene. Gene. 1980 Jul;10(2):157–166. doi: 10.1016/0378-1119(80)90133-x. [DOI] [PubMed] [Google Scholar]
  14. Vrsanská M., Biely P., Krátký Z. Enzymes of the yeast lytic system produced by Arthrobacter GJM-1 bacterium and their role in the lysis of yeast cell walls. Z Allg Mikrobiol. 1977;17(6):465–480. doi: 10.1002/jobm.3630170608. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES