SYNTHESIS AND APPLICATION OF SORBENTS FOR AFFINITY-CHROMATOGRAPHY OF SERINE PROTEASES

A synthetic peptide derivative, - alanyl-alanyl-leucyl-morpholide, (Al a-Ala-Leu-N(CH2CH2)(2)O), is suggested as a specific Ligand for affini ty chromatography of subtilisin-like serine proteases that prefer hydr ophobic amino acid residue in P1 position of their substrates. Z-Ala-A la-Leu-N(CH2CH2), a weak and reversible inhibitor of serine proteases, was synthesized by the carbodiimide method. Affinity sorbents were pr epared by coupling the synthesized peptide derivates to CH or AH Sepha rose. Serine proteases from different sources were purified up to 17 f old on these sorbents with yields varying from 25 % to 100 %. Three en zymes (serine protease X, kallikrein and leucine aminopeptidase) were isolated from urine of children with glomerulonephritis with yields of 57, 22 and 55 %. Proteolytic enzymes from the dandelion root, Kamchat ka crab and culture filtrates of different microorganisms were also pu rified on the affinity sorbents.