METHOD OF COMMERCIAL E.coli STRAIN CULTURE PREPARED OF BL21(DE3) STRAIN CARRYING POLYMERASE T7 RNA GENE UNDER CONTROL OF lacUV5 PROMOTER WITH INCREASED BIOMASS SYNTHESIS AND HIGHER END PROTEIN YIELD IN INCLUSION BODIES

摘要

FIELD: medicine.;SUBSTANCE: method under the invention provides growing of an inoculate on the basis of BL21(DE3) E. coli strain carrying polymerase T7 RNA gene under control of lacUV5 promoter to the middle of a logarithmic phase of growth on a nutrient medium with the use of glucose in the concentration of 10 g/l as a carbon source at temperature 38°C, agitation at 200 rpm for 3 h; a fermenter is sown with the inoculate at a sowing dose of 10%; the microorganisms are cultured at 38°C, the dissolved oxygen concentration of 80% at maximum mixing rate 1200 rpm, maximum aeration level 1 volume of air /min: 1.2 volumes of a culture fluid, pH 7.00; termination of fermentation after 12-13 hours of culture; deposition of the cells of the E. coli producing strain. At the beginning of culture, 50% (wt %) glucose is added in the middle of the logarithmic phase of growth, 80% (wt %) glycerol is added in portions and the logarithmic phase of culture growth is terminated by fractional additions of 25% (wt %) lactose. After culture, the biomass containing recombinant end proteins in the form of insoluble inclusion bodies is deposited.;EFFECT: invention enables higher yield of the biomass and the end protein of a volume unit of the culture medium.;8 dwg, 4 ex