Chemistry: molecular biology and microbiology – Micro-organism – tissue cell culture or enzyme using process... – Recombinant dna technique included in method of making a...
Patent
1996-01-16
1998-08-04
Ketter, James
Chemistry: molecular biology and microbiology
Micro-organism, tissue cell culture or enzyme using process...
Recombinant dna technique included in method of making a...
435 711, 435171, 4351723, 4352542, 43525421, 4352546, 4352556, 4352543, 4352567, 536 231, C12P 2106
Patent
active
057891931
DESCRIPTION:
BRIEF SUMMARY
FIELD OF THE INVENTION
This invention relates to recombinant-DNA-technology. Specifically this invention relates to new recombinant eukaryotic cells transformed with SSO genes or their homologues. A eukaryotic cell transformed with several copies of a SSO gene or a gene homologous to SSO has an increased capacity to produce secreted foreign or endogenous proteins.
Further, the said new recombinant eukaryotic cells, especially yeasts and filamentous fungi, when transformed with genes expressing suitable hydrolytic enzymes can hydrolyze and/or utilize appropriate macromolecular/polymeric compounds more efficiently, which results in increased cell mass production and/or more versatile utilization of the compounds in relevant biotechnical applications.
BACKGROUND OF THE INVENTION
The development of recombinant DNA methods has made it possible to produce proteins in heterologous host systems. This possibility greatly facilitates production of e.g. proteins of therapeutic importance which normally occur in nature in very low amounts or are otherwise difficult to isolate or purify. Such proteins include growth factors, hormones and other biologically active proteins or peptides which traditionally have been isolated from human or animal tissues or body fluids e.g. blood serum or urine. The increasing danger of the presence of human pathogenic viruses such as HBV, HIV, and oncogenic viruses or other pathogens in the human or animal tissues or body fluids has greatly speeded up the search for heterologous production systems for these therapeutics. Other proteins of clinical importance arc viral or other microbial or human parasite proteins needed for diagnostics and for vaccines especially of such organisms which are difficult to grow in vitro or in tissue culture, or are dangerous human pathogens. These include viruses like HBV, HIV, yellow fever, rubella, FMDV, rabies, and human parasites such as malaria.
A further group of proteins for which heterologous production systems have been or are being developed are secreted enzymes, especially those hydrolyzing plant material, and which are needed in food and fodder production as well as in other industrial processes including textile industry and pulp and paper industry. The possibility of producing proteins in heterologous systems or production of endogenous proteins in genetically engineered cells increases their yields and greatly facilitates their purification and has already by now had a great impact on studies of structure and function of many important enzymes and other proteins. The production and secretion of foreign hydrolytic enzymes in yeast for example, results in improvements in processes based on industrial yeast strains such as distiller's, brewer's or baker's yeasts.
Various production systems have been and are being developed including bacteria, yeasts, filamentous fungi, animal and plant cell cultures and even multicellular organisms like transgenic animals and plants. All of these different systems have their advantages, even if disadvantages, and all of them are needed.
The yeast Saccharomyces cerevisiae is at the moment the best known eukaryote at genetic level. As a eukaryotic microbe it possesses the advantages of a eukaryotic cell like most if not all of the post-translational modifications of eukaryotes, and as a microbe it shares the easy handling and cultivation properties of bacteria. The large scale fermentation systems are well developed for S. cerevisiae which has a long history as a work horse of biotechnology including production of food ingredients and beverages such as beer and wine.
The yeast genetic methods are by far the best developed among eukaryotes based on the vast knowledge obtained by classical genetics. This made it easy to adopt and further develop for yeast the gene technology procedures first described for Escherichia coli. Along other lines the methods for constructing yeast strains producing foreign proteins have been developed to a great extent (Romanos et. al., 1992).
Secretion of the proteins into the culture
REFERENCES:
Mohamed, A.H., et al. 1988. Journal of Biological Chemistry. vol. 263. No. 25, pp. 12315-12325. "Primary Structure of the Multisubunit . . . ".
Mohamed, A. H., et al. 1989. Gen Bank Accession No. J03936.
Bennett, M.K, et al. Science vol. 257, 10 Jul. 1992. pp. 255-259. "Syntaxin: A Synaptic Protein Implizated in Docking . . . ".
Inoue, et al., "Cloning and Sequence Analysis of cDNA for a Neuronal Cell Membrane Antigen, HPC-1", The Journal of Biological Chemistry 267(15):10613-10619, (May 1992).
Martegani, et al., "Expression of high levels of human tissue plasminogen activator in yeast under the control of an inducible GAL promoter", Applied Microbiology and Biotechnology 37:604-608, (1992).
Hirai, et al., "Epimorphin: A Mesenchymal Protein Essential for Epithelial Morphogenesis", Cell 69:471-481 (May 1, 1992).
Aalto et al., "Yeast syntaxins Sso1p and Sso2p belong to a family of related membrane proteins that function in vesicular transport," EMBO J., 12(11):4095-4104(1993).
Bennett et al., "The molecular machinery for secretion is conserved from yeast to neurons, " Proc. Nat'l Acad. Sci., USA, 90:2559-2563 (1993).
Case et al., "Efficient transformation of Neurospora crassa by utilizing hybrid plasmid DNA," Proc. Nat'l Acad. Sci., USA, 76(10):5259-5263 (Oct., 1979).
Ferro-Novick et al., Vesicle fusion from yeast to man, Nature, 370:191-193 (Jul. 21, 1994).
Jeenes et al., "Heterologous Protein Production by Filamentous Fungi," Biotechnology and Genetic Reviews, 9:327-367 (Dec., 1991).
Penttila et al., "A versatile transformation system for the cellulolytic filamentous fungus Trichoderma reesei," Gene, 61:155-164 (1987).
Sudhof et al., "Membrane Fusion Machinery: Insights from Synaptic Proteins," Cell, 75:1-4 (Oct. 8, 1993).
Tilburn et al., "Transformation by integration in Aspergillus nidulans," Gene, 26:205-221 (1983).
Aalto Markku
Keranen Sirkka
Outola Mika
Penttila Merja
Ronne Hans
Ketter James
Valtion teknillinen tutkimuskeskus
Yucel Irem
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