Chemistry: molecular biology and microbiology – Micro-organism – tissue cell culture or enzyme using process... – Enzymatic production of a protein or polypeptide
Patent
1985-04-12
1988-05-24
Martinell, James
Chemistry: molecular biology and microbiology
Micro-organism, tissue cell culture or enzyme using process...
Enzymatic production of a protein or polypeptide
435 70, 435 71, 4351721, 4351723, 435253, 435320, 435811, 935 33, 935 34, 935 38, 935 43, 935 61, 935 73, C12P 2100, C12P 2102, C12P 2104, C12N 1500, C12N 120, C12N 100
Patent
active
047466081
DESCRIPTION:
BRIEF SUMMARY
TECHNICAL FIELD
The present invention relates to a process for producing physiologically active peptides derived from eukaryotes, for example, .beta.-interferon (hereinafter referred to as .beta.-IFN), by fermentation.
BACKGROUND ART
Recently, genes coding for useful, physiologically active peptides derived from eukaryotes, for example, .beta.-IFN have been cloned and linked to promoters derived from microorganisms to efficiently produce the desired physiologically active peptides in microorganisms by utilizing rapidly developed genetic engineering techniques [T. Taniguchi, et al.: Proc. Jap. J. Acad. sec B 55, 464-469 (1979), Japanese Published Unexamined Patent Application No. 77654/82, Japanese Published Unexamined Patent Application No. 110600/83].
DISCLOSURE OF THE INVENTION
A technique of efficiently culturing microorganisms harboring recombinant plasmids according to their properties and producing desired physiologically active peptides in a high yield by fermentation is very important.
Generally, the optimum conditions for producing proteins and enzymes by microbial cells need not always be identical with the optimum growth conditions for the cells themselves, and greatly depend on culturing conditions such as culturing temperature, pH, and aeration-agitation and medium conditions. The production process also greatly depends on whether the desired protein or enzyme is of induction type or of constitution type. For example, in the case of induction type, the production amount greatly varies with the conditions for adding an inducer, etc. When the desired protein or enzyme is unstable, it is necessary to study the process for stable production.
The present inventors have studied various culturing conditions described above for producing the desired, physiologically active peptides in a high yield by fermentation, using microorganisms having recombinant plasmids and capable of producing .beta.-IFN, etc. As a result, the present inventors have successfully found the conditions for highly efficient production of physiologically active peptides such as .beta.-IFN, and have established the present invention.
The present invention is described in detail below.
The present invention provides a process for producing a peptide by culturing a microorganism containing a recombinant DNA comprising a gene coding for the peptide derived from eukaryotes, a vector and a promoter and being capable of producing the peptide in a medium, accumulating the peptide in the culture broth, and recovering the peptide from the culture broth, characterized by conducting the culturing at a temperature 10.degree. to 25.degree. C. lower than the optimum growth temperature for the microorganism.
Up to now, various physiologically active peptides have been expressed in microorganisms, and in most cases, Escherichia coli has been used as the microorganisms. In the case of Escherichia coli, the culturing is usually carried out at a temperature of 37.degree. C., and actual production of these physiologically active peptides is mostly carried out at 37.degree. C. In accordance with the present invention, peptides can be efficiently produced by culturing at a temperature 10.degree. to 25.degree. C. lower, preferably 15.degree. to 20.degree. C. lower than the ordinary culturing temperature. For example, in the case of Escherichia coli, a good result can be obtained by culturing at 15.degree. to 30.degree. C., preferably at 20.degree. to 25.degree. C.
In the present invention, peptides such as insulin, interferon, growth hormone, etc., preferably human .beta.-interferon peptide, can be mentioned as the peptides derived from eukaryotes. As the vector, vectors derived from Escherichia coli, vectors derived from Bacillus subtilis, and vectors derived from yeasts, preferably pBR322, pBR313 and pMB9 derived from Escherichia coli, can be used. As the promoter, lac promoter, phoA promoter, trp promoter, etc., preferably trp promoter can be used.
As the medium for the present process, a medium usually used for culturing host microorganisms c
REFERENCES:
Taniguchi et al, Proc. Natl. Acad. Sci. USA 77: 5230 (1980).
Hallewell et al, Gene 9: 27 (1980).
Herendeen et al, J. Bacteriol. 139(1), 185 (1979).
Patent Abstracts of Japan, vol. 7, No. 259, 58-141796, 8/23/83.
Ito Seiga
Mizukami Tamio
Nishi Tatsunari
Oka Tetsuo
Kyowa Hakko Kogyo Co. Ltd.
Martinell James
LandOfFree
Process for producing peptides does not yet have a rating. At this time, there are no reviews or comments for this patent.
If you have personal experience with Process for producing peptides, we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and Process for producing peptides will most certainly appreciate the feedback.
Profile ID: LFUS-PAI-O-1058176