Skip to main content
PMC home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1983 Aug;155(2):657–663. doi: 10.1128/jb.155.2.657-663.1983

The penicillinase of Bacillus licheniformis is an outer membrane protein in Escherichia coli.

M O Sarvas, I A Palva
PMCID: PMC217736  PMID: 6223920

Abstract

The cloned gene coding for Bacillus licheniformis penicillinase (penP) was introduced into Escherichia coli in a heat-inducible lambda Qam vector. After induction, significant amounts of penicillinase were synthesized in the new host. The cellular location of the penicillinase was found to be almost exclusively the outer membrane fraction of E. coli, and virtually no soluble penicillinase was found. According to sodium dodecyl sulfate-gel electrophoresis, the size of the penicillinase from E. coli was identical to that of the membrane-bound form of the B. licheniformis penicillinase. Gel filtration in the presence of Triton X-100 suggested that the penicillinase from E. coli had amphiphilic properties, as does B. licheniformis membrane penicillinase. These results show that the export of the penicillinase to the outer membrane of E. coli involves the cleavage of the signal peptide from the prepenicillinase, giving an outer membrane component indistinguishable from the membrane penicillinase of B. licheniformis.

Full text

PDF
657

Images in this article

661Image
on p.661
662Image
on p.662

Selected References

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

  1. Brammar W. J., Muir S., McMorris A. Molecular cloning of the gene for the beta-lactamase of Bacillus licheniformis and its expression in Escherichia coli. Mol Gen Genet. 1980 Apr;178(1):217–224. doi: 10.1007/BF00267232. [DOI] [PubMed] [Google Scholar]
  2. Chang C. N., Nielsen J. B., Izui K., Blobel G., Lampen J. O. Identification of the signal peptidase cleavage site in Bacillus licheniformis prepenicillinase. J Biol Chem. 1982 Apr 25;257(8):4340–4344. [PubMed] [Google Scholar]
  3. Cornelis P., Digneffe C., Willemot K. Cloning and expression of a Bacillus coagulans amylase gene in Escherichia coli. Mol Gen Genet. 1982;186(4):507–511. doi: 10.1007/BF00337957. [DOI] [PubMed] [Google Scholar]
  4. Fraser T. H., Bruce B. J. Chicken ovalbumin is synthesized and secreted by Escherichia coli. Proc Natl Acad Sci U S A. 1978 Dec;75(12):5936–5940. doi: 10.1073/pnas.75.12.5936. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Gray O., Chang S. Molecular cloning and expression of Bacillus licheniformis beta-lactamase gene in Escherichia coli and Bacillus subtilis. J Bacteriol. 1981 Jan;145(1):422–428. doi: 10.1128/jb.145.1.422-428.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Hantke K., Braun V. Covalent binding of lipid to protein. Diglyceride and amide-linked fatty acid at the N-terminal end of the murein-lipoprotein of the Escherichia coli outer membrane. Eur J Biochem. 1973 Apr;34(2):284–296. doi: 10.1111/j.1432-1033.1973.tb02757.x. [DOI] [PubMed] [Google Scholar]
  7. Ichihara S., Hussain M., Mizushima S. Characterization of new membrane lipoproteins and their precursors of Escherichia coli. J Biol Chem. 1981 Mar 25;256(6):3125–3129. [PubMed] [Google Scholar]
  8. Imanaka T., Tanaka T., Tsunekawa H., Aiba S. Cloning of the genes for penicillinase, penP and penI, of Bacillus licheniformis in some vector plasmids and their expression in Escherichia coli, Bacillus subtilis, and Bacillus licheniformis. J Bacteriol. 1981 Sep;147(3):776–786. doi: 10.1128/jb.147.3.776-786.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Izui K., Nielsen J. B., Caulfield M. P., Lampen J. O. Large exopenicillinase, initial extracellular form detected in cultures of Bacillus licheniformis. Biochemistry. 1980 Apr 29;19(9):1882–1886. doi: 10.1021/bi00550a023. [DOI] [PubMed] [Google Scholar]
  10. Lai J. S., Sarvas M., Brammar W. J., Neugebauer K., Wu H. C. Bacillus licheniformis penicillinase synthesized in Escherichia coli contains covalently linked fatty acid and glyceride. Proc Natl Acad Sci U S A. 1981 Jun;78(6):3506–3510. doi: 10.1073/pnas.78.6.3506. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. MALAMY M., HORECKER B. L. The localization of alkaline phosphatase in E. coli K12. Biochem Biophys Res Commun. 1961 Jun 2;5:104–108. doi: 10.1016/0006-291x(61)90020-1. [DOI] [PubMed] [Google Scholar]
  12. Michaelis S., Beckwith J. Mechanism of incorporation of cell envelope proteins in Escherichia coli. Annu Rev Microbiol. 1982;36:435–465. doi: 10.1146/annurev.mi.36.100182.002251. [DOI] [PubMed] [Google Scholar]
  13. Neu H. C., Heppel L. A. The release of enzymes from Escherichia coli by osmotic shock and during the formation of spheroplasts. J Biol Chem. 1965 Sep;240(9):3685–3692. [PubMed] [Google Scholar]
  14. Neugebauer K., Sprengel R., Schaller H. Penicillinase from Bacillus licheniformis: nucleotide sequence of the gene and implications for the biosynthesis of a secretory protein in a Gram-positive bacterium. Nucleic Acids Res. 1981 Jun 11;9(11):2577–2588. doi: 10.1093/nar/9.11.2577. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Nielsen J. B., Caulfield M. P., Lampen J. O. Lipoprotein nature of Bacillus licheniformis membrane penicillinase. Proc Natl Acad Sci U S A. 1981 Jun;78(6):3511–3515. doi: 10.1073/pnas.78.6.3511. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Nielsen J. B., Lampen J. O. Glyceride-cysteine lipoproteins and secretion by Gram-positive bacteria. J Bacteriol. 1982 Oct;152(1):315–322. doi: 10.1128/jb.152.1.315-322.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. 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]
  18. Osborn M. J., Gander J. E., Parisi E., Carson J. Mechanism of assembly of the outer membrane of Salmonella typhimurium. Isolation and characterization of cytoplasmic and outer membrane. J Biol Chem. 1972 Jun 25;247(12):3962–3972. [PubMed] [Google Scholar]
  19. Sanderson K. E., Ross H., Ziegler L., Mäkelä P. H. F + , Hfr, and F' strains of Salmonella typhimurium and Salmonella abony. Bacteriol Rev. 1972 Dec;36(4):608–637. doi: 10.1128/br.36.4.608-637.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Sarvas M., Hirth K. P., Fuchs E., Simons K. A precursor form of the penicillinase from Bacillus licheniformis. FEBS Lett. 1978 Nov 1;95(1):76–80. doi: 10.1016/0014-5793(78)80055-6. [DOI] [PubMed] [Google Scholar]
  21. Simons K., Sarvas M., Garoff H., Helenius A. Membrane-bound and secreted forms of penicillinase from Bacillus licheniformis. J Mol Biol. 1978 Dec 25;126(4):673–690. doi: 10.1016/0022-2836(78)90015-3. [DOI] [PubMed] [Google Scholar]
  22. Smith W. P., Tai P. C., Davis B. D. Bacillus licheniformis penicillinase: cleavages and attachment of lipid during cotranslational secretion. Proc Natl Acad Sci U S A. 1981 Jun;78(6):3501–3505. doi: 10.1073/pnas.78.6.3501. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Talmadge K., Stahl S., Gilbert W. Eukaryotic signal sequence transports insulin antigen in Escherichia coli. Proc Natl Acad Sci U S A. 1980 Jun;77(6):3369–3373. doi: 10.1073/pnas.77.6.3369. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Yamamoto S., Lampen J. O. The hydrophobic membrane penicillinase of Bacillus licheniformis 749/C. Characterization of the hydrophilic enzyme and phospholipopeptide produced by trypsin cleavage. J Biol Chem. 1976 Jul 10;251(13):4102–4110. [PubMed] [Google Scholar]

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

RESOURCES