METHOD FOR OBTAINING RECOMBINANT BETA-1,4-GALACTOSIDASE BGAA FROM STREPTOCOCCUS PNEUMONIAE

摘要

The invention relates to the field of biotechnology and addresses the problem of obtaining industrial quantities of highly purified enzymatically active recombinant β-1,4-galactosidase BgaA which is suitable for use in the remodeling stage to obtain therapeutically functional glycoprotein preparations with specified properties, for developing tools for high-sensitivity analysis of glycan chains of glycoproteins of plant, animal and microbial tissue. A method involves the use of metal chelate, pseudo-affinity, hydrophobic, anion-exchange, concentrating chromatography with subsequent polishing gel filtration, wherein a protein from the soluble fraction of cell lysate is purified using two stages of chromatography on the same Ni2+ metal chelate sorbent under different elution conditions and by pseudo-affinity chromatography in flow-through mode on Blue Sepharose. The use of the three stages mentioned above helps achieve maximum purification of the preparation for major contaminating impurities.