Hemostatically active proteases of the micromycete Aspergillus protuberus: formation and properties

Abstract

Hemostatically active proteases of the micromycete Aspergillus protuberus are of considerable interest for the development of new drugs regulating blood clotting. The introduction substantiates the relevance of the study related to the need for natural compounds for the correction of hemostasis disorders. The purpose of this work is to study the biochemical properties, synthesis conditions, and medical potential of A. protuberus proteases. The materials and methods section describes approaches to optimizing mushroom cultivation (variation of carbon sources, pH, aeration), multi-stage purification of enzymes (ultrafiltration, ion exchange chromatography), as well as analysis methods (mass spectrometry, kinetic tests, in vitro models). The results demonstrate that the maximum production of proteases is achieved when glucose and casein are used in a medium with a pH of 6.0–7.0. The unique structural features of the enzymes, including glycosylation sites, provide a specific interaction with fibrinogen and thrombin. In preclinical models, purified proteases reduce bleeding time by 40%, but raw extracts exhibit a synergistic effect. The discussion highlights the prospects for the use of proteases in biotechnology and medicine, but notes the risks of systemic effects on hemostasis. Ways of increasing specificity (genetic modification) and stability (immobilization on nanoparticles) are proposed. The conclusions emphasize the need for further safety research, scaling up production, and developing standardized dosage forms. The work contributes to the understanding of the role of microbial proteases in the regulation of coagulation and opens up directions for the creation of innovative hemostatic drugs.