Abstract
PurposeIn this study, an aqueous two-phase micellar system (ATPMS), formed by the non-ionic surfactant Triton X-114, was used to investigate the partitioning of cellulolytic enzymes produced by the filamentous fungus Aspergillus fumigatus CCT 7873.MethodsPerformance of the ATPMS on the partitioning of CMCase (activity on carboxymethyl cellulose) and FPase (activity on filter paper) was investigated by varying the temperature (35, 40, 45, 50, 55, 60, and 65 °C), enzyme crude extract concentration (20, 40, 60, and 80% w/w), and Triton X-114 concentration (2, 4, 6, and 8% w/w) and by adding different inorganic salts (NaCl, CaCl2, MgSO4, and MnSO4) in the system.ResultsAn ATPMS formed with 8% (w/w) Triton X-114 and 40% (w/w) enzymatic crude extract at a system temperature of 55 °C was most favorable for partitioning the tested enzymes. Under these conditions, a purification factor for CMCase and FPase of 10.89 and 0.65 was reached, respectively. The addition of inorganic salts changed the distribution of enzymes. Of these, CaCl2 contributed to a higher distribution coefficient (50.0), whereas for FPase, the presence of MnSO4 in the system improved the purification factor to 3.94.ConclusionThe highest values obtained for the yield and purification factors demonstrate that ATPMS is an interesting option for recovering and purifying cellulolytic enzymes.
Highlights
Brazil is a large producer of agro-industrial products, and it generates a lot of lignocellulosic residues (Machado et al 2010)
The aqueous two-phase micellar systems (ATPMS) is based on the use of a surfactant in order to form two immiscible phases, a bottom phase that is rich in micelles and an upper phase that is poor in micelles
Due to the importance of reducing the costs of downstream processing by exploiting integrative techniques, this study investigated the partitioning behavior of cellulases using an ATPMS formed by a nonionic surfactant (Triton X-114), inorganic salts, and crude extract at different concentrations
Summary
Brazil is a large producer of agro-industrial products, and it generates a lot of lignocellulosic residues (Machado et al 2010) These residues are rich in cellulose, hemicelluloses, and lignin. Integrative techniques play a key role in downstream processing One such technique involves aqueous two-phase micellar systems (ATPMS). The formation of micelles is related to the equilibrium of intermolecular forces, including Van der Waals, hydrophobic, steric, and electrostatic ones (Liu et al 1998) Once these selfassembling aggregates are achieved, a biphasic system is built (Liu et al 1996), which can be exploited for biomolecule separation/purification. The main surfactant used is a nonionic one, such as Triton X-114 (Wang et al 2013)
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