Purification of α-galactosidase and invertase by three-phase partitioning from crude extract of Aspergillus oryzae

Abstract

Alpha-galactosidase and invertase were accumulated in a coherent middle phase in a three-phase partitioning system under different conditions (ammonium sulphate, ratio of tert-butanol to crude extract, temperature and pH). Alpha-galactosidase and invertase were purified 15- and 12-fold with 50 and 54% activity recovery, respectively. The fractions of interfacial precipitate arising from the three-phase partitioning were analyzed by SDS-PAGE. Both purified preparations showed electrophoretic homogeneity on SDS-PAGE.

Changes in the Contents of Sugars and Organic Acids during the Ripening and Storage of Sufu, a Traditional Oriental Fermented Product of Soybean Cubes

“…Catalytic hydrolysis of stachyose, raffinose, and sucrose by the hydrolytic enzymes such as R-galactosidase and β-fructosidase/ invertase produced by Asp. oryzae extracellularly during the fermentation might lead to this result (14,15). After being kept at room temperature for another month, the sufu was found to contain 1.72, 376.78, 261.75, and 9.12 mg/g of dried matter of maltose, glucose, fructose, and galactose, respectively.…”

Three-phase partitioning as a rapid and efficient method for purification of invertase from tomato

“…Quite some successful applications of TPP have been described. By using TPP technique several enzymes and proteins have been purified with different purification folds and yields [10][11][12][13][14][18][19][20]25,26].…”

“…It has been used both for upstream and downstream protein purification processes and has sometimes been used as a one step purification protocol [10,11,17]. TPP based separation is also very simple, quick and inexpensive process in comparison to conventional purification processes [10,17,18]. TPP has been used to purify a number of biomolecules such as; protease inhibitor from ragi [11], soybean trypsin inhibitor [12], excreted-secreted protein fraction from Corynebacterium pseudotuberculosis [13], ␣-galactosidase and invertase from Aspergillus oryzae [18], pectinase from tomato [19], phospholipase D from Dacus carota [20], xylanase from Aspergillus niger [21].…”

Partial purification of invertase from Momordica charantia (bitter melon) by three phase partitioning (TPP) method

“…Since the method was first reported in 1972, more than 100 research studies have been published on the use of TPP separation technology for the extraction and purification of a variety of biomolecules including proteins, enzymes, enzyme inhibitors, edible oils or lipids, carbohydrates and small-molecule organic compounds from diverse natural sources (Yan et al, 2017). Presently, invertase has been purified from five plant sources by the TPP methodology (Lovrein et al, 1987;Dhananjay and Mulimani, 2008;Kat and Yılmazer, 2013).…”

“…The methods used to purify different enzymes from other sources may not be suitable for a novel enzyme or source, because enzymes can be affected by various factors (ammonium sulfate concentration, ratio of tbutanol, pH and incubation time) in the method. Therefore, the method must be optimized, hence in the case of enzyme purification with TPP, the method has been optimized in the literature Chaiwut et al, 2010;Dhananjay and Mulimani, 2008;Duman and Kaya, 2013;Gagaoua et al, 2014;Hussain et al, 2009;Liu et al, 2006, Sagu et al, 2015.…”