Vázquez-Villegas Patricia, Espitia-Saloma Edith, Torres-Acosta Mario A, Ruiz-Ruiz Federico, Rito-Palomares Marco, Aguilar Oscar
Escuela de Ingenieria y Ciencias, Tecnologico de Monterrey, Monterrey, Mexico.
Escuela de Medicina y Ciencias de la Salud, Tecnologico de Monterrey, Monterrey, Mexico.
Front Chem. 2018 Oct 2;6:454. doi: 10.3389/fchem.2018.00454. eCollection 2018.
Aqueous two-phase systems (ATPS) have been reported as an attractive biocompatible extraction system for recovery and purification of biological products. In this work, the implementation, characterization, and optimization (operational and economic) of invertase extraction from spent brewery yeast in a semi-automatized pilot plant using ATPS is reported. Gentian violet was used as tracer for the selection of phase composition through phase entrainment minimization. Yeast suspension was chosen as a complex cell matrix model for the recovery of the industrial relevant enzyme invertase. Flow rates of phases did not have an effect, given that a bottom continuous phase is given, while load of sample and number of agitators improved the recovery of the enzyme. The best combination of factors reached a recovery of 129.35 ± 2.76% and a purification factor of 4.98 ± 1.10 in the bottom phase of a PEG-Phosphate system, also resulting in the removal of inhibitor molecules increasing invertase activity as reported by several other authors. Then, an economic analysis was performed to study the production cost of invertase analyzing only the significant parameters for production. Results indicate that the parameters being analyzed only affect the production cost per enzymatic unit, while variations in the cost per batch are not significant. Moreover, only the sample load is significant, which, combined with operational optimization results, gives the same optimal result for operation, maximizing recovery yield (15% of sample load and 1 static mixer). Overall res ults of these case studies show continuous pilot-scale ATPS as a viable and reproducible extraction/purification system for high added-value biological compounds.
水相两相系统（ATPS）已被报道为一种用于生物制品回收和纯化的具有吸引力的生物相容性提取系统。在这项工作中，报告了在半自动化中试工厂中使用ATPS从废啤酒酵母中提取转化酶的实施、表征和优化（操作和经济方面）。使用结晶紫作为示踪剂，通过使相夹带最小化来选择相组成。选择酵母悬浮液作为用于回收工业相关酶转化酶的复杂细胞基质模型。鉴于有底部连续相，相流速没有影响，而样品负载和搅拌器数量提高了酶的回收率。在聚乙二醇 - 磷酸盐系统的底部相中，最佳因素组合实现了129.35±2.76%的回收率和4.98±1.10的纯化因子，还如其他几位作者所报道的那样导致抑制剂分子的去除从而提高了转化酶活性。然后，进行了经济分析，仅分析生产的重要参数来研究转化酶的生产成本。结果表明，所分析的参数仅影响每酶单位的生产成本，而每批次成本的变化不显著。此外，只有样品负载是显著的，结合操作优化结果，得到相同的最佳操作结果，使回收率最大化（样品负载的15%和1个静态混合器）。这些案例研究的总体结果表明，连续中试规模的ATPS是一种用于高附加值生物化合物的可行且可重复的提取/纯化系统。
