A Novel Aqueous Micellar Two-Phase System Composed of Surfactant and Sorbitol for Purification of Pectinase Enzyme fromPsidium guajavaand Recycling Phase Components

Abstract

A novel aqueous two-phase system composed of a surfactant and sorbitol was employed for the first time to purify pectinase from Psidium guajava. The influences of different parameters, including the type and concentration of the surfactant and the concentration and composition of the surfactant/sorbitol ratio, on the partitioning behavior and recovery of pectinase were investigated. Moreover, the effects of system pH and the crude load on purification fold and the yield of purified pectinase were studied. The experimental results indicated that the pectinase was partitioned into surfactant-rich top phase, and the impurities were partitioned into the sorbitol-rich bottom phase with the novel method involving an ATPS composed of 26% (w/w) Triton X-100 and 23% (w/w) sorbitol at 54.2% of the TLL crude load of 20% (w/w) at pH 6.0. The enzyme was successfully recovered by this method with a high purification factor of 15.2 and a yield of 98.3%, whereas the phase components were also recovered and recycled at rates above 96%. This study demonstrated that this novel ATPS method can be used as an efficient and economical alternative to the traditional ATPS for the purification and recovery of the valuable enzyme.

Extraction of lipase fromRhizopus microsporusfermentation culture by aqueous two-phase partitioning

“…The amount of added crude broth to the ATPS is important because the loaded feedstock can change the partitioning behaviour of the target protein and the phase V R . [19] Similar to the results of Amid et al [40] on pectinase partitioning, the increasing amounts of both lipase and contaminants in the systems could result in a decrease in the ATPS performance. ATPS experiments were carried out by varying the loaded fermentation broth to 40% (w/w).…”

Analyzing the effects of protecting osmolytes on solute–water interactions by solvatochromic comparison method: I. Small organic compounds

“…The dominant hypothesis is based on the preferential solvation model, according to which osmolytes are excluded from protein surface and increase the Gibbs free energy change associated with protein unfolding. [10][11][12][13][14][15][16][17][18][19] The recently reported observation that sorbitol may form aqueous two-phase system in mixture with Triton X-100, 20 in our view, serves as an unambiguous indication that sorbitol does change the structure and properties of water. This view is based on the observations that the aqueous solubility of non-polar amino acids and peptides decreases in solutions of osmolytes relative to that in pure water.…”

Purification of a Bacteriocin‐Like Inhibitory Substance Derived from Pediococcus acidilactici Kp10 by an Aqueous Micellar Two‐Phase System

“…The AMTPS was composed of 22.5% w/w TX‐100 and 50% w/w CFCS. The concentrations of the additives were chosen based on the work of previous researchers . The mixtures were incubated at 75 °C for 30 min for the biphasic formation.…”

“…Previously, the use of Triton X‐114, a nonionic surfactant, in AMTPS for the recovery of bacteriocin have been reported by several researchers . Furthermore, Triton X‐100 (TX‐100) has been used in AMTPS for the recovery of pectinase from kesinai plant ( Streblus asper ) leaves . Other nonionic surfactant, n‐decyltetraethylene oxide (C 10 E 4 ) has also been used in the extraction of clavulanic acid from a Streptomyces clavuligerus fermentation broth …”

“…To our knowledge, this is the first report on the use of TX‐100‐based AMTPS for BLIS recovery. TX‐100 was chosen for the system because it is the most commonly used nonionic surfactant in research laboratory and therefore can be easily obtainable, and is relatively the cheapest compared to other nonionic surfactants …”