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利用双水相体系(PEG/磷酸盐)和磁场提取和纯化具有转果糖基活性的β-呋喃果糖苷酶。

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

机构信息

Federal University of Agreste of Pernambuco, Garanhuns, Brazil.

Education Unit of Penedo, Federal University of Alagoas, Penedo, Brazil.

出版信息

Prep Biochem Biotechnol. 2022;52(4):478-486. doi: 10.1080/10826068.2021.1964085. Epub 2021 Aug 24.

DOI:10.1080/10826068.2021.1964085
PMID:34428129
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 URM4634 was extracted using PEG/Phosphate Aqueous Biphasic Systems (ABS), and the impact of magnetic field on the extraction behavior was evaluated. A 2-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 (), purification factor (), activity yield () and selectivity (). 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 ( = 0.50,  = 4.05,  = 72.66% and  = 0.06) were observed at pH 8.0 using 12.5% (w/w) PEG 400 and 25% (w/w) NaHPO/KHPO. 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 ( = 0.93,  = 4.22,  = 83.79% and  = 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)参与蔗糖水解和低聚果糖的生产,这对食品工业具有重要意义。使用聚乙二醇/磷酸盐双水相系统(ABS)提取 URM4634 的 FFase,并评估磁场对提取行为的影响。采用 2 全实验设计研究了聚乙二醇的摩尔质量、聚乙二醇和磷酸盐的浓度以及 pH 对所选响应变量(分配系数()、纯化因子()、活性产率()和选择性())的影响。还研究了磁场在分配过程中的影响和 NaCl 浓度对相同响应的影响。在没有磁场的情况下,FFase 提取的最佳结果( = 0.50、  = 4.05、  = 72.66%和  = 0.06)是在 pH 8.0 下使用 12.5%(w/w)PEG 400 和 25%(w/w)NaHPO/KHPO 观察到的。磁场的应用可以提高性能,在磁铁之间距离最长(磁场最低)且没有 NaCl 的情况下获得最佳结果( = 0.93、  = 4.22、  = 83.79%和  = 0.09)。所得结果表明,ABS 与低强度磁场结合可用于 FFase 的预纯化。

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