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.

摘要

β-呋喃果糖苷酶 (FFases) 是参与蔗糖水解和低聚果糖生产的酶，对食品工业具有重要意义。使用 PEG/磷酸盐双相系统 (ABS)提取Aspergillus tamarii URM4634 中的 FFase，并评估了磁场对提取行为的影响。采用 2 ⁴全实验设计，研究了 PEG 摩尔质量、PEG 和磷酸盐浓度以及 pH 值对所选响应变量即分配系数 ( K )、纯化因子 ( PF )、活性产率 ( Y ) 的影响。 ) 和选择性 ( S）。还研究了分区期间磁场和NaCl浓度对相同响应的影响。使用 12.5% (w/w) PEG 400 和 25% (w/w) 在 pH 8.0 下观察到无磁场 FFase 提取的最佳结果 ( K  = 0.50, PF  = 4.05, Y  = 72.66% 和S  = 0.06) NaH ₂ PO ₄ /K ₂ HPO ₄。磁场的应用可以提高性能，在磁铁之间的最长距离（最低磁场）和不存在 NaCl（K  = 0.93，PF  = 4.22，Y  = 83.79% 和S = 0.09)。所获得的结果表明，ABS 与低强度磁场的组合可用作一种有效的 FFase 预纯化方法。