A Novel Aqueous Two Phase System Composed of Surfactant and Xylitol for the Purification of Lipase from Pumpkin (Cucurbita moschata) Seeds and Recycling of Phase Components

Abstract

Lipase is one of the more important enzymes used in various industries such as the food, detergent, pharmaceutical, textile, and pulp and paper sectors. A novel aqueous two-phase system composed of surfactant and xylitol was employed for the first time to purify lipase from Cucurbita moschata. The influence of different parameters such as type and concentration of surfactants, and the composition of the surfactant/xylitol mixtures on the partitioning behavior and recovery of lipase was investigated. Moreover, the effect of system pH and crude load on the degree of purification and yield of the purified lipase were studied. The results indicated that the lipase was partitioned into the top surfactant rich phase while the impurities partitioned into the bottom xylitol-rich phase using an aqueous two phase system composed of 24% (w/w) Triton X-100 and 20% (w/w) xylitol, at 56.2% of tie line length (TLL), (TTL is one of the important parameters in this study and it is determined from a bimodal curve in which the tie-line connects two nodes on the bimodal, that represent concentration of phase components in the top and bottom phases) and a crude load of 25% (w/w) at pH 8.0. Recovery and recycling of components was also measured in each successive step process. The enzyme was successfully recovered by the proposed method with a high OPEN ACCESSMolecules 2015, 20 11185 purification factor of 16.4 and yield of 97.4% while over 97% of the phase components were also recovered and recycled. This study demonstrated that the proposed novel aqueous two phase system method is more efficient and economical than the traditional aqueous two phase system method for the purification and recovery of the valuable enzyme lipase.

Liquid Biphasic System: A Recent Bioseparation Technology

“…An [58]. In addition, the recyclability of both components in surfactant-LBS extraction makes this an excellent process, as this innovative method was practical and feasible to be applied in the biotechnology industry for extraction of other biomolecules.…”

Purification and Characterization of Lipase Produced by Leuconostoc mesenteroides Subsp. mesenteroides ATCC 8293 Using an Aqueous Two-Phase System (ATPS) Composed of Triton X-100 and Maltitol

“…It has been reported that non-ionic surfactant such as Triton X-100 induces lipase activity, and therefore, reduces enzyme denaturation. Besides, Triton X-100 could also increase enzyme activity by affecting the enzyme-substrate interaction, in which, the addition of this non-ionic surfactant prevented the adsorbed enzyme from inactivation [ 5 , 7 ]. However, at 50% ( w / w ) Triton X-100 concentration, lipase activity started to decrease as higher concentrations of surfactants negatively affected the amount of solubilized enzyme and its catalytic action.…”

“…However, these two traditional systems have some drawbacks such as the use of salt in the system might lead to the denaturation or certain target proteins may precipitate due to the high ionic-strength environment [ 4 ]. Furthermore, additional laborious purification techniques, such as ultrafiltration, diafiltration and crystallization are necessary for the removal of phase-forming chemicals/polymers from the target protein [ 5 ]. In order to overcome the weaknesses of the traditional ATPS systems, new ATPS systems composed of non-ionic surfactants and polyols have been discovered and explored [ 6 , 7 ].…”

Aqueous two-phase extraction technique based on magnetic ionic liquid-nonionicsurfactant for phenolic acids in grape, apple, pear, and banana

“…For this, the amount of Triton X-100 was optimized for this study. On the other hand, low or high concentrations of surfactant negatively influenced the amount of solubilized phenolic acids [30]. The recoveries were decreased after 0.2 g of Triton X-100 in the extractions repeated with three ionic liquids.…”