METHOD OF PRODUCING RECOMBINANT Β-1,4-GALACTOSIDASE BGAA FROM STREPTOCOCCUS PNEUMONIAE

摘要

FIELD: biotechnology.;SUBSTANCE: invention relates to biotechnology and solves the problem of obtaining highly purified enzymatically active recombinant β-1,4-galactosidase BgaA, suitable for use at the remodeling step for producing therapeutically functional preparations of glycoproteins with given properties, to develop tools for high-sensitivity analysis of glycan chains of plant, animal and microbial glycoproteins. Method comprises using metal chelate, pseudo-affine, hydrophobic, anion-exchange, concentrating chromatography followed by polishing gel filtration, wherein protein from soluble fraction of cell lysate is purified by two chromatography steps on same Ni²⁺ metal chelate sorbent under different elution conditions and pseudo-affine chromatography in breakthrough mode on Blue Sepharose.;EFFECT: use of said three steps helps to maximize purity of the preparation from major contaminating impurities.;10 cl, 8 dwg, 3 tbl, 6 ex