Extração de proteases fibrinolíticas de Auricularia a. delicata utilizando sistema de duas fases aquosas - SDFA formado por PEG/sulfato de sódio

Abstract

Cardiovascular diseases have been the main cause of death in the world. Thrombosis is one such disease and it happens when fibrin builds up in the arteries. According to the World Health Organization (WHO), about 17.7 million people will die this year from cardiovascular diseases, and in 2030, that amount will be 23.6 million. In view of the great potential of the Amazonian microbial biodiversity and the increasing applicability of enzymes in the production of medicines, this research aims to evaluate the production capacity of fibrinolytic proteases by Auricularia A.delicata. For this purpose, extractive fermentation techniques and enzyme purification by a biphasic aqueous PEG/sodium sulfate system were used. In the recovered extracts, tests of in vitro degradation of the blood clot were performed by the fibrin plaque methods and quantification by spectrophotometer. The fermented product was subjected to extraction and recovery by an aqueous two-phase system. Partition coefficient (K), purification factor (FP) and enzyme recovery were determined. The highest fibrinolytic activity (30.71 U/mL) was obtained in the phase rich in sodium sulfate in the assay composed of 10% salt and 18% PEG 8000 (g/mol). Recoveries greater than 80% were obtained. As a result, the process is shown to be effective to pre-purify fibrinolytic proteases at low cost and significant speed. When compared to other isolated production and purification techniques, extractive fermentation is an environmentally friendly process and can replace conventional initial separation steps.