Ivanko O. Collagenase and keratinase of Streptomycetes

The dissertation is focused on purification and characterization of collagenase and keratinase of streptomycetes. As result of screening among 364 strains of microorganisms of different taxonomy groups (micromycetes, yeasts, bacilli and streptomycetes) it were found two producers of collagenase and keratinase - Streptomyces sp. 1349 and Streptomyces sp. 1382, respectively. The analysis of the heterogeneity of Streptomyces sp. 1349 population allowed to reveal the colony of basic type with activity that exceeds 2,3 times that of the a wild type. This level of activity is stable during some generations of culture. It has been shown that NH4Cl and soybean meal increase the collagenolytic activity of Streptomyces sp. 1349 in 5,0-6,0 fold, and sheep's wool or chicken feathers (as the only source of N and C) increase keratinolytic activity of Streptomyces sp. 1382 in 7,0 fold. Three collagenases and two keratinases were obtained from culture liquids of strain-producers. It was found two collagenases are metalloproteases, and other collagenase and keratinases are serine proteases. Physical and chemical properties, specificity of action of purified enzymes were investigated. It has been shown that preparations of enzymes are differ in their molecular weights, composition of amino acids, optimal conditions of their action, sensitivity for cations of metals and anions. Collagenases are characterized by wide specificity to different protein substrates, while keratinases are able to hydrolyze only native keratin of wool or feathers. Also, one of keratinase has high fibrinolytic activity. It has been shown that collagenases of Streptomyces sp. 1349 have no serological affinity for enzyme for C. histolyticum ("Merk"). For the first time it has been shown that certain coordinating substances of germanium, guanidine derivatives and inhibitors of virus proteolysis inhibit collagenase and keratinase activities of Streptomyces sp. 1349 and Streptomyces sp. 1382, respectively. It could be used during creation of new effectivedrugs for treatment of skin disease caused by collagenases and keratinases of microorganisms.