Purification of potato polyphenol oxidase (PPO) by partitioning in aqueous two-phase system

Partition and purification of a thermostable xylanase produced by Paecilomyces thermophila in solid-state fermentation using aqueous two-phase systems

“…As an alternative to the first purification step, the fermentation broth could be subjected to a purification process based on ATPS [12,25]. ATPS has shown to be useful for the primary purification of enzymes; however, the enzymes obtained by this method frequently contain impurity [5][6][7][8]12,14]. Gaikaiwari et al used ATPS to separate xylanase, although they reached only a purification factor of about 2.6 with a 98% enzyme recovery [5].…”

“…The use of ATPS in downstream processing has also performed on the extraction, separation, concentration and primary purification of various enzymes that include xylanase, xylose reductase, b-glucosidase, potato polyphenol oxidase, etc. [5][6][7][8]. As ATPS can remove contaminants such as nucleic acids and undesirable proteins, it has been successfully used to purify b-xylosidase and lysozyme to near homogeneity [9,10].…”

Process integration for the recovery and purification of recombinant Pseudomonas fluorescens proline dehydrogenase using aqueous two-phase systems

“…The degradation of proline to glutamate is dependent on the multifunctional proline utilization A (PutA) flavoprotein, a peripherally membrane-bound enzyme that includes proline dehydrogenase (ProDH; EC1.5.99.8) and D 1 -pyrroline-5-carboxylate dehydrogenase (P5CDH; EC1.5.1.12) domains [2]. ProDH is a flavoenzyme with a (␤␣) 8 barrel catalytic core that catalyzes the conversion of proline to 1 -pyrroline-5-carboxylate (P5C) in the presence of flavin adenine dinucleotide (FAD) as a cofactor [3]. Hyperprolinemia type I (HPI) and schizophrenia are attributed to the deficiency in ProDH gene.…”

“…In ATPS processes, the differential separation is driven by van der waals, hydrophobic, hydrogen bond, and ionic interactions between the phase forming components. Therefore, the partition is influenced by the concentration and molecular mass of polymer and salt, ionic strength, temperature, pH and also the physiochemical characteristics of desired biomolecule [7,8]. We have already isolated a ProDH enzyme from Pseudomonas fluorescens bacterium, and succeeded in gene cloning and its recombinant production in Escherichia coli [5].…”

Extraction and Purification of Lipoxygenase from Soybean Using Aqueous Two-Phase System

“…Therefore, there is a need for alternative approaches to the problem. In this regard, aqueous two-phase extraction (ATPE) provides a powerful method for separating and purifying mixtures of proteins (Vaidya et al 2006). ATPE has been widely used for the extraction and purification of many biological products due to their technical simplicity, ease of scaleup and high yields (Raghavarao et al 1995).…”