METHOD OF PRODUCING BIOCATALYST EXHIBITING CEPHALOSPORIN ACID SYNTHESIS ACTIVITY Russian patent published in 2011 - IPC C12N11/04 

FIELD: medicine.

SUBSTANCE: method provides cultivation of Escherichia coli cells producing cephalosporin acid synthetase. As selection criteria of the moment of enzyme biosynthesis truncation point, the pH value of a cultural fluid and a enzyme distribution constant C_dist=A_cbm/A_p-p, where A_cbm is enzymatic adaptation of a cell biomass (Me/g humid) and A_p.p-is enzymatic adaptation of a supernatant (ME/ml). The cephalosporin acid synthetase biosynthesis is truncated at C_dist being not less than 350 and pH value 7.9÷8.2. The produced cell biomass in the concentration of 100÷150 mg dry/g of a polymerisation mixture is suspended in a solution of acrylamide and N, N'-methylene-bis-acrylamide in the weight relation 20:1. During 5÷30 min, the cells are modified by glutaric aldehyde taken in the relative concentration of 1·10^-3÷9·10^-3 mmol/mg of protein. The polymerisation process is initiated by persulfuric acid salt. Synthetase activity of the biocatalyst is 750-950 MU/g dry.

EFFECT: method allows producing the biocatalyst which exhibits high target activity and stability and does not contain proteins washed out in operation of proteins contaminating the biocatalytic transformation end-product.

3 tbl, 7 ex