Neira-Vielma Alberto A, Iliná Anna, Álvarez Georgina Michelena, Nascimento Cynthia O, Aguilar Cristóbal Noé, Martínez-Hernández José Luis, Carneiro-da-Cunha Maria das Graças
Food Research Department, Universidad Autónoma de Coahuila, México. Blvd. V. Carranza S/N. Col. República, CP 25280, Saltillo, Coahuila, México.
Departamento de Bioquímica/Laboratório de Imunopatologia Keizo Asami, Universidade Federal de Pernambuco-UFPE, Av. Prof. Moraes Rego s/n, CEP 50.670-420, Recife, PE, Brazil.
Biotechnol Rep (Amst). 2020 May 20;26:e00471. doi: 10.1016/j.btre.2020.e00471. eCollection 2020 Jun.
This work describes the successful extraction of Aspergillus niger phytase from a crude extract (CE) obtained from solid-state fermentation by reversed micelle system using anionic surfactant sodium bis (2-ethylhexyl) sulfosuccinate (AOT) in isooctane achieved in two simple steps: forward and backward extractions. The effects of potassium chloride (KCl) concentration, pH of the aqueous solution, and AOT concentration that affect the system were examined. The best result for the forward extraction was obtained with the CE solution at pH 4.0, 50 mM KCl, and 100 mM AOT, while for the backward extraction the best result was achieved with a stripping aqueous solution at pH 5.5 containing 200 mM KCl, achieving a purification factor of 4.03, 1.15 times higher than that reported for the conventional purification process. Phytase purity was demonstrated by SDS-PAGE (89 kDa) and its activity by zymogram, confirming the efficiency of the process with low time consumption (∼40 min).
本研究描述了通过反胶束体系从固态发酵获得的粗提物（CE）中成功提取黑曲霉植酸酶的过程。该反胶束体系使用异辛烷中的阴离子表面活性剂双（2-乙基己基）磺基琥珀酸钠（AOT），通过两个简单步骤实现：正向萃取和反向萃取。研究了氯化钾（KCl）浓度、水溶液pH值和AOT浓度对该体系的影响。正向萃取的最佳结果是在pH 4.0、50 mM KCl和100 mM AOT的CE溶液中获得，而反向萃取的最佳结果是在pH 5.5、含有200 mM KCl的反萃水溶液中实现，纯化因子达到4.03，比传统纯化工艺报道的结果高1.15倍。通过SDS-PAGE（89 kDa）证明了植酸酶的纯度，通过酶谱证明了其活性，证实了该工艺效率高且耗时短（约40分钟）。
