resumo
Aqueous two-phase systems (ATPS) based on tetrahydrofuran (THF) + potassium phosphate buffer (pH 7) were used in this work for the purification of lipases. Binodal curve, tie lines and critical point were obtained for the new THF-salt ATPS at 25 degrees C and the binodal curve were successfully correlated with the Merchuck and Hu equations. To optimize the extraction capability of this ATPS the effects of the concentration of components and temperature of equilibrium on the partition coefficients and extraction efficiencies were investigated using lipase from Burkholderia cepacia (commercially obtained) as a model compound. The optimum conditions for the purification of an extracellular lipase obtained by submerged fermentation were established through a surface response analysis by central composite rotational design applied allowing a purification factor (PF) of 103.9 +/- 0.9 and an enzyme recovery of 96.4 +/- 1.1 achieved using this process. Moreover, a commercial lipase by Candida antarctica B recombinant in Aspergillus niger was purified (PF= 4.84 +/- 0.24), confirming the potential of this new THF-based ATPS for purifying lipases. (C) 2015 Elsevier Ltd. All rights reserved.
palavras-chave
PORCINE PANCREATIC LIPASE; POLYETHYLENE-GLYCOL; PHASE-SEPARATION; IONIC LIQUIDS; INDUSTRIAL APPLICATIONS; PROTEIN SEPARATION; BIPHASIC SYSTEMS; TOLERANT LIPASE; PLUS WATER; EXTRACTION
categoria
Biochemistry & Molecular Biology; Biotechnology & Applied Microbiology; Engineering
autores
Souza, RL; Lima, RA; Coutinho, JAP; Soares, CMF; Lima, AS
nossos autores
agradecimentos
The authors acknowledge the financial support of FAPITEC/SE, CAPES and CNPq. The authors also thank financial support through the doctoral grant R.L. Souza and PROBIC/UNIT for the scholarship for R.A. Lima. J.A.P. Coutinho acknowledges the Fundacao para a Ciencia e a Tecnologia for the funding of CICECO-Aveiro Institute of Materials (Ref. FCT UID/CTM/50011/2013).