FIELD: biotechnology.
SUBSTANCE: invention relates to biotechnology and a method for industrial cultivation of a recombinant strain E.coli BL21 (DE3)/pMD704 – a producer of penicillin G acylase. Method provides production of ECOPGA preparation in industrially significant amounts. Method provides growing of inoculum based on E.coli BL21 (DE3)/pMD704 strain to middle of logarithmic phase of growth on nutrient medium using glucose as carbon source at temperature of 38 °C, stirring at 200 rpm for 3–5 hours, seed transfer fermenter with inoculum, further cultivation of strain in fermenter at 22–23 °C on medium containing sources of nitrogen, carbon, inorganic salts, with concentration of dissolved oxygen of 80–90 % at maximum mixing rate of 1,200 rpm, maximum level of aeration 1 volume of air/min : 1 volume of culture fluid. After cultivation, the produced biomass is precipitated and destroyed. End product (ECOPGA) is extracted in the form of an enzyme paste by precipitation with the help of polyethylene glycol.
EFFECT: invention increases output of biomass and functional-active penicillin G acylase from unit volume of culture medium; as a result, a preparation ECOPGA is obtained, which is suitable for producing biocatalysts for synthesis of cephalosporin acids.
1 cl, 3 dwg, 1 tbl, 4 ex
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