Method and system for enhanced production of commercially import

Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving virus or bacteriophage

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435 71, 435 691, 435 693, 4353201, 435190, 435193, 435209, 435201, 435233, 435234, 435471, 43525231, 4352523, 435 698, C12Q 168, C12N 121, C12N 1567, C12N 1575

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059452781

ABSTRACT:
The invention provides a method and expression system for enhancing secretion of hyperproduced homologous and heterologous exoproteins in gram-positive bacteria such as Bacillus sp. The method and system comprise overproduction of PrsA protein in a gram-positive bacterial host also overproducing at least one exoprotein of interest. Use of the method and system of the invention results in greatly enhanced secretion of the synthesized exoproteins into the growth medium. Once in the growth medium these secreted exoproteins can be recovered and purified in a straightforward manner.

REFERENCES:
patent: 5780261 (1998-07-01), Kontinen et al.
Wickner, W., Driessen, A.J.M., and Hartl, F. The Enzymology of Protein Translocation Across the Escherichia coli Plasma Membrane, Annual Reviews of Biochemistryl , 60: 101-124 (1989).
Kontinen, V. and Sarvas, M. Mutants of Bacillus subtilis Defective in Protein Export. Journal of General Microbiology 134: 2333-2344 (1988).
Kontinen, V. and Sarvas, M. A Gene (prsA) of Bacillus subtilis involved in a novel, late stage of protein export. Molecular Microbiology 5(5): 1273-1283 (1991).
Haandrikman, A.J., et al. Identification of a Gene Required or Maturation of an Extracellular Lactococcal Serine Proteinase. Journal of Bacteriology 171(5): 2789-2794 (1989).
Vos, P. et al. A Maturation Protein is Essential for Production o Active Forms of Lactococcus lactis SK11 Serine Proteinase Located in or Secreted from the Cell Envelope. Journal of Bacteriology 171(5): 2795-2802 (1989).
Ferrari, F.A., Nguyen, A., Lang, D. and Hoch, J.A. Construction and Properties of an Integrable Plasmid for Bacillus subtilis. Journal of Bacteriology 154(3): 1513-1515 (1983).
Sibakov, M., Sarvas, M. and Palva, I. Increased secretion of .alpha.-amylase from Bacillus subtilis caused by multiple copies of .alpha.-amylase Gene from B. amyloliquefaciens is not further increased by genes enhancing the basic level of secretion. FEMS Microbiology Letters 17: 81-85 (1983).
Gardel, C., Johnson, K., Jacq, A. and Beckwith, J. The secD locus of E. coli codes for two membrane proteins required for protein export. EMBO Journal 9(10): 3209-3216 (1990).

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