Extraction and purification of Aspergillus tamarii β-fructofuranosidase with transfructosylating activity using aqueous biphasic systems (PEG/phosphate) and magnetic field

Abstract

β-fructofuranosidases (FFases) are enzymes involved in sucrose hydrolysis and fructo-oligosaccharides’ production which are of great interest for the food industry. FFase from Aspergillus tamarii URM4634 was extracted using PEG/Phosphate Aqueous Biphasic Systems (ABS), and the impact of magnetic field on the extraction behavior was evaluated. A 2⁴-full experimental design was employed to study the influence of molar mass of PEG, concentrations of PEG and phosphate and pH on the selected response variables, i.e., partition coefficient (K), purification factor (PF), activity yield (Y) and selectivity (S). The influence of magnetic field during partition and NaCl concentration on the same responses was also studied. The best results of FFase extraction without magnetic field (K = 0.50, PF = 4.05, Y = 72.66% and S = 0.06) were observed at pH 8.0 using 12.5% (w/w) PEG 400 and 25% (w/w) NaH₂PO₄/K₂HPO₄. Application of the magnetic field allowed improving the performance, with the best results being obtained at the longest distance between magnets (lowest magnetic field) and absence of NaCl (K = 0.93, PF = 4.22, Y = 83.79% and S = 0.09). The outcomes obtained demonstrate that ABS combination with low intensity magnetic field can be used as an efficient FFase pre-purification method.

Keywords:

• Aqueous biphasic system
• β-fructofuranosidase
• magnetic field
• purification

Additional information

Funding

The authors acknowledge the financial support of Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq, Brazil) (Finance code: 471773/2013-1) and Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES, Brazil) (Finance code: 23038.003634/2013-15), and Federal Rural University of Pernambuco (UFRPE, Brazil). R. L. de Oliveira is especially grateful to Fundacão de Amparo à Ciência e Tecnologia do Estado de Pernambuco (FACEPE, Brazil) for the post-doctoral scholarship (grant BFP-0105-5.07/20).