Liquid–liquid extraction of lipase produced by psychrotrophic yeast Leucosporidium scottii L117 using aqueous two-phase systems

Abstract

a b s t r a c tAqueous two-phase systems (ATPS) have been used in biomolecules separation and as an efficient alternative to traditional purification systems for lipases extraction. Here, we investigated the partitioning and recovery of lipase derived from Leucosporidium scottii L117 using ATPS and aqueous two-phase micellar systems (ATPMS). Thus, we evaluated three ATPS: (i) polyethylene glycol (PEG)/phosphate salts and (ii) PEG/polyacrylic acid (NaPA) in different molecular weights (1500, 4000 and 8000 g/mol). (iii) Triton X-114 (TX-114)/McIlvaine buffer pH 7.0 in different conditions (2.0% (w/w) of TX-114 at 25.0 and 28.0°C). The PEG/phosphate and PEG/NaPA systems resulted in a great loss of enzymatic activity; thus these systems do not represent viable alternatives for these lipase extraction. The micellar systems yielded the best results for lipase extraction with enzyme activity balances ranging between 84.7% and 113.05%. After optimizing the micellar system by experimental design of the partition coefficient of lipase increased by 10.3-fold (0.75-7.76). Lipase preferentially partitioned into the micelle-rich phase with K Lip = 7.76, %REC Bot = 93.85% and PF = 1.2 at 25.03°C, 5.1 pH and 10.38% TX-114 and K Lip = 4.77, % REC Bot = 73.53% and PF = 1.97 at 28.00°C, 4.5 pH and 8.0% TX-114, indicating that the ATPMS represents an alternative to purification/extraction of lipase L. scottii L117. A crude lipase extract was also evaluated to define the optimum pH and temperature. Lipase reached optimal activity at 40°C, and remained stable in pH values ranging from pH 3.0 to 8.0 and temperatures from 20.0 to 45.0°C, with relative residual lipase activity above 80% after 30 min of incubation.

Yeasts from macroalgae and lichens that inhabit the South Shetland Islands, Antarctica

“…Most of the filamentous fungi and yeast that occurs in Antarctica are cosmopolitan, some were transported to Antarctica by migratory animals, while others called indigenous, are well adapted to the Antarctic environment, and are able to grow and reproduce even at low temperatures (Ruisi et al, ). Survival strategies that have been observed in cold‐adapted yeasts recovered from Antarctic environment, including production of antifreeze proteins (Hashim et al ., ), cold shock and heat shock adaptation (Boo et al ., ), pigment with photoprotective activity (Vaz et al ., ), and cold‐active enzymes (Carrasco et al ., ; Duarte et al ., ).…”

Biosurfactant production by yeasts from different types of soil of the South Shetland Islands (Maritime Antarctica)

“…; Duarte et al . ) and for bioremediation purposes (Buzzini et al . ), they could present potential for the production of specific metabolites, such as biosurfactants.…”

“…According to Buzzini et al (2012), 70 species of yeasts isolated from Antarctica were identified, 13 being ascomycetes and 57 basidiomycetes. Besides their exploitation for the production of coldactive enzymes (Buzzini et al 2012;Duarte et al 2015) and for bioremediation purposes (Buzzini et al 2012), they could present potential for the production of specific metabolites, such as biosurfactants.…”

Purification of bacterial inulinase in aqueous two‐phase systems

“…Therefore, efficient methods for recuperation and purification of fermentation products such as enzymes and biologically active proteins are in demand. These systems are composed of two matters soluble in water that are immiscible beyond critical concentration [8][9][10][11][12]. These systems are composed of two matters soluble in water that are immiscible beyond critical concentration [8][9][10][11][12].…”

“…An alternative method for separation of biomolecules based on aqueous two -phase systems (ATPSs) was proposed by Albertsson, 1985 [7]. These systems are composed of two matters soluble in water that are immiscible beyond critical concentration [8][9][10][11][12]. Varieties of water soluble maters are used for obtaining ATPS.…”