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From the highly purified but lowly active recombinant protein Destabilas-Lysozyme (Dest-Lys) by use cation-exchange column TSK CM 3-SW chromatography, it was separated non-active fraction IV, contained 90% of protein. Fractions I, II and III, represented proteins with lysozyme and isopeptidase activities. Their lysozyme activity correlates with the activity of natural Des-Lys. The ratio of the activities in fractions I - III is such, that maximal lysozyme activity is concentrated in fraction III, isopeptidase - in fraction I. It is discussed the possibility of Dest-Lys different functions regulation is depended on the formation of protein complex forms.

The protective antithrombotic effects of piyavit, a novel pharmacological preparation from the medicinal leech, its water extract concentrated and diluted 1600-fold, the piyavit fraction containing leech prostanoids and highly purified destabilase, epsilon-(gamma-Glu)-Lys-isopeptidase isolated from the medicinal leech (Hirudo medicinalis), have been studied. All the preparations under study inhibited clot formation in rat mesanterium arterioles stimulated by laser energy, contained leech prostanoids immunoenzymatically identified as 6-keto-PgF1 alpha and inhibited platelet aggregation induced by ADP. The putative mechanism of their antithrombotic effect is discussed.

Piyavit, the novel pharmacological preparation from the medicinal leech (Hirudo medicinalis) contained the leech saliva, produces the potent arterial antithrombotic effect examined on experimental of Laser Induced Thrombus formation. Administrated orally into rats or injected subcutaneously as water extract, non-diluted or in 1600 times diluted, it inhibits statistically significant comparing with control platelet thrombus stimulated by laser beams. Its components, prostanoid fraction and purified enzyme destabilase, endo-epsilon-(gamma-Glu)-Lys-isopeptidase, also inhibit thrombus formation in statistically different manner, comparing with control. All the tested preparations inhibit platelet aggregation induced by ADP. The dependence of arterial antithrombotic effect on the leech prostanoid is discussed.

Catalytic activity of destabilase has been studied in hydrolyses of synthetic and natural substrates. High specificity of the enzyme is shown in hydrolysis of endo-epsilon-(gamma-Glu)Lys isopeptide bonds.