Recovery and partitioning of fibrinolytic protease from Bacillus sp. UFPEDA 485 by aqueous two-phase systems

Abstract

Fibrinolytic proteases produced by Bacillus sp. has attracted interest in the pharmaceutical industry as a promising alternative in thrombolytic therapy due to their effectiveness in degrading fibrin, its production requiring the development of an efficient recovery process. Aqueous two-phase systems (ATPS) have been recognized as an efficient and economical process for recovering enzymes. To optimize the recovery of fibrinolytic protease from the fermentation broth of Bacillus sp. UFPEDA 485, a 23 full factorial design was used to evaluate the influence of the three independent variables PEG molar mass (MPEG), PEG concentration (CPEG) and sodium sulfate concentration (CNa2SO4) on the partition coefficient (K), purification factor (PF) and yield recovery (Y) of fibrinolytic protease in PEG/Na2SO4 aqueous two-phase system. For all ATPS studied, enzymes partitioned to the top phase and the highest extraction was obtained for MPEG 6000 g.mol-1, CPEG 24 % (w/w) and CNa2SO4 11.6 % (w/w) with K = 5.03; PF = 3.30; Y = 91.40% and Fibrinolytic activity in the top phase 821 U.mL-1. Findings reported here show that ATPS composed of PEG/Na2SO4 is a valuable strategy for the extraction of fibrinolytic protease and can be considered a promising method for the extraction of enzymes in industrial scale.