Recovery and purification of Aspergillus niger phytase from crude extract using AOT / isooctane reversed micelles

• Published in: Biotechnology reports (Amsterdam, Netherlands) Vol. 26; p. e00471
• Main Authors: 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
• Format: Journal Article
• Language: English
• Published: Elsevier B.V 01.06.2020; Elsevier
• Subjects: Liquid-liquid extraction; Phytase; Reversed micelles; Triticale; Liquid-liquid extraction; Phytase; Reversed micelles; Triticale
• ISSN: 2215-017X; 2215-017X
• DOI: 10.1016/j.btre.2020.e00471

[Display omitted] •The application of the reverse micelles resulted in purification of A. niger phytase.•It was possible purify phytase from A. niger by reversed micelles in short period time.•Reversed micelles proved to be a viable alternative for phytase purification.•Phytase remained active after extraction using AOT/isooctane reversed micelles.•The phytase purity and activity were confirmed by SDS-PAGE and zymogram analyzes. 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).