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A Gene Coding for a Membrane–Bound Hydrolase is Expressed as a Secreted, Soluble Enzyme in Streptomyces Lividans

Bio/Technology volume 7, pages 65–68 (1989)Cite this article

Abstract

A Flavobacterium opd gene, which encodes for a membrane–associated phosphotriesterase, was cloned into Streptomyces lividans strain 66. The gene product was constitutively produced in the streptomycete as a soluble, extracellular enzyme. S. lividans (Gram–positive) was able to use the Flavobacterium (Gram–negative) promoter region for high level expression of the enzyme. This cloning system may be of value for the expression and secretion of other membrane–associated proteins

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References

  1. McLaughlin, J.R., Murray, C.L., and Rabinowitz, J.C. 1981. Unique features in the ribosome binding site sequence of the gram-positive Staphylococcus aureus beta-lactamase gene. J.Biol.Chem. 256:11283–11291.

    CAS  PubMed  Google Scholar 

  2. Rosenberg, M. and Court, D. Regulatory sequences involved in the promotion and termination of RNA transcription. 1979. Ann. Rev. Genet. 13:319–353.

    Article  CAS  PubMed  Google Scholar 

  3. Moran, Jr. C.P., Lang, N., LeGrice, S.F.J., Lee, G., Stephens, M., Sonenshein, A.L., Pero, J. and Losick, R. 1982. Nucleotide sequences that signal the initiation of transcription and translation in Bacillus subtilis. Mol. Gen. Genet. 186:339–346.

    Article  CAS  PubMed  Google Scholar 

  4. Ehrlich, S.D. and Sgaramella, V. 1978. Barriers to the heterospecific gene expression among procaryotes. Trends in Biochem.Sci. 3:259–261.

    Article  CAS  Google Scholar 

  5. Schottel, J.L., Bibb, M.J. and Cohen, S.N. 1981. Cloning and expression in Streptomyces lividans of antibiotic resistance genes derived from Eschenchia coli. J.Bacteriol. 146:360–368.

    CAS  PubMed  PubMed Central  Google Scholar 

  6. Bibb, M.J. and Cohen, S.N. 1982. Gene expression in Streptomyces: construction and application of promoter-probe plasmid vectors in Streptomyces lividans. Mol. Gen. Genet. 187:265–277.

    Article  CAS  PubMed  Google Scholar 

  7. Bernan, V., Filpula, D., Herber, W., Bibb, M. and Katz, E. 1985. The nucleotide sequence of the tyrosine gene from Streptomyces antibioticus and characterization of the gene product. Gene 37:101–110.

    Article  CAS  PubMed  Google Scholar 

  8. Eckhardt, T., Strickler, J., Gorniak, L., Burnett, W.V. and Fare, L.R. 1987. Characterization of the promoter, signal sequence, and amino terminus of a secreted β-galactosidase from “Streptomyces lividans”. J. Bacteriol. 169:4249–4256.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  9. Brown, K.A. 1980. Phosphotriesterases of Flavobacterium sp. Soil Biol.Biochem. 12:105–112.

    Article  CAS  Google Scholar 

  10. McDaniel, C.S., Harper, L.L. and Wild, J.R. 1988. Cloning and sequencing of a plasmid-borne gene (opd) encoding a phosphotriesterase. J.Bacteriol. 170:2306–2311.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  11. Serdar, C.M., Gibson, D.T., Munnecke, D.M., and Lancaster, J.H. 1982. Plasmid involvement in parathion hydrolysis by Pseudomonas diminuta. Appl. Environ. Microbiol. 44:246–249.

    CAS  PubMed  PubMed Central  Google Scholar 

  12. Mulbry, W.W., Karns, J.S., Kearney, P.C., Nelson, J.O., McDaniel, C.S. and Wild, J.R. 1986. Identification of a plasmid-borne parathion hydrolase gene from Flavobacterium sp.by Southern hybridization with opd from Pseudomonas diminuta. Appl. Environ. Microbiol. 51:926–930.

    CAS  PubMed  PubMed Central  Google Scholar 

  13. Kearney, P.C., Karns, J.S., Muldoon, M.T., and Ruth, J.M. 1986. Coumaphos disposal by combined microbial and UV-ozonation reactions. J. Agric. Food Chem. 34:702–706.

    Article  CAS  Google Scholar 

  14. Kearney, P.C., Karns, J.S. and Mulbry, W.W. 1987. Engineering soil microorganisms for pesticide degradation, p. 591–596. In: Pesticide Science and Biotechnology. R. Greenhalgh and T. R. Roberts (Eds.).Blackwell Scient.Publ., London.

    Google Scholar 

  15. Gauger, W.K., MacDonald, J.M., Adrian, N.R., Matthees, D.P. and Walgenbach, D.D. 1986. Characterization of a streptomycete growing on organophosphate and carbamate insecticides. Arch. Environ. Contam. Toxicol. 15:137–141.

    Article  CAS  PubMed  Google Scholar 

  16. Hopwood, D.A., Bibb, M.J., Chater, K.F., Kieser, T., Bruton, C.J., Kieser, H.M., Lydiate, D.J., Smith, C.P., Ward, J.M. and Schrempf, H. 1985. Genetic manipulation of Streptomyces. A laboratory manual. F.Crowe and Sons, Ltd., Norwich, England.

    Google Scholar 

  17. Serdar, C.M. and Gibson, D.T. 1985. Enzymatic hydrolysis of or-ganophosphates: cloning and expression of a parathion hydrolase gene from Pseudomonas diminuta. Bio/Technology 3:567–571.

    CAS  Google Scholar 

  18. Maniatis, T., Fritsch, E.F. and Sambrook, J. 1982. Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, N.Y.

    Google Scholar 

  19. Laemmli, U.K. 1970. Cleavage of structural proteins during assembly of the head of bacteriophage T4. Nature 227:680–685.

    Article  CAS  PubMed  Google Scholar 

  20. Henderson, G., Krygsman, P., Liu, C.J., Davey, C.C. and Malek, L.T. 1987. Characterization and structure of genes for proteases A and B from Streptomyces griseus. J. Bacteriol. 169:3778–3784.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  21. Vieira, J. and Messing, J. 1982. The pUC plasmids, an M13mp7-derived system for insertion mutagenesis and sequencing with synthetic universal primers. Gene 19:259–268.

    Article  CAS  PubMed  Google Scholar 

  22. Holmes, D.S. and Quigley, M. 1981. A rapid boiling method for the preparation of bacterial plasmids. Anal. Biochem. 114:193–197.

    Article  CAS  PubMed  Google Scholar 

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Author notes

  1. John G. Steiert

    Present address: Department of Microbiology, University of Iowa, Iowa City, Iowa, 52242

  2. Burton M. Pogell: Corresponding author.

Authors and Affiliations

  1. Department of Medicinal Chemistry/Pharmacognosy, School of Pharmacy, University of Maryland, Baltimore, Maryland, 21201

    Burton M. Pogell, Marilyn K. Speedie & James Laredo

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  1. John G. Steiert
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  2. Burton M. Pogell
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  4. James Laredo
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Steiert, J., Pogell, B., Speedie, M. et al. A Gene Coding for a Membrane–Bound Hydrolase is Expressed as a Secreted, Soluble Enzyme in Streptomyces Lividans. Nat Biotechnol 7, 65–68 (1989). https://doi.org/10.1038/nbt0189-65

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