Novel aqueous two-phase systems based on tetrahydrofuran and potassium phosphate buffer for purification of lipase. Issue 9 (September 2015)

• Record Type: Journal Article
• Authors: Souza, Ranyere L.; Lima, Rafaella A.; Coutinho, João A.P.; Soares, Cleide M.F.; Lima, Álvaro S.
• Is Part Of: Process biochemistry. Volume 50:Issue 9(2015:Sep.)
• Page Start: 1459
• Page End: 1467
• Publication Date: 2015-09
• Subjects: Aqueous biphasic systems -- Tetrahydrofuran -- Phosphate buffer -- Purification -- Lipase; Biochemical engineering -- Periodicals; Biotechnology -- Periodicals; Biochemistry -- periodicals; Biotechnology -- periodicals; Chemical Engineering -- periodicals; Génie biochimique -- Périodiques; Biotechnologie -- Périodiques; Biochemical engineering; Biotechnology; Periodicals; 660.63
• DOI: 10.1016/j.procbio.2015.05.015
• Languages: English
• ISSNs: 1359-5113

Abstract

Graphical abstract: Highlights: Novel ATPS composed of THF and K2 HPO4 /KH2 PO4 at pH 7 was studied. The phase diagram, tie-lines and critical point were determined at 298 K. The concentration of components and temperature of equilibrium was investigated. The enzyme partitioning is spontaneous and governed by entropic effects. Lipase from Bacillus produced by submerged fermentation was purified 103.9 (fold). Abstract: 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 °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.