Song Cher Pin, Liew Poh En, Teh Zora, Lim Schian Pei, Show Pau Loke, Ooi Chien Wei
Chemical Engineering Discipline, School of Engineering, Selangor, Malaysia.
Advanced Engineering Platform, School of Engineering, Monash University Malaysia, Selangor, Malaysia.
Front Chem. 2018 Oct 31;6:529. doi: 10.3389/fchem.2018.00529. eCollection 2018.
The formation of aqueous two-phase system (ATPS) with the environmentally friendly and recyclable ionic liquid has been gaining popularity in the field of protein separation. In this study, the ATPSs comprising ,-dimethylammonium ''-dimethylcarbamate (DIMCARB) and thermo-responsive poly(propylene) glycol (PPG) were applied for the recovery of recombinant green fluorescent protein (GFP) derived from . The partition behavior of GFP in the PPG + DIMCARB + water system was investigated systematically by varying the molecular weight of PPG and the total composition of ATPS. Overall, GFP was found to be preferentially partitioned to the hydrophilic DIMCARB-rich phase. An ATPS composed of 42% (w/w) PPG 1000 and 4.4% (w/w) DIMCARB gave the optimum performance in terms of GFP selectivity (1,237) and yield (98.8%). The optimal system was also successfully scaled up by 50 times without compromising the purification performance. The bottom phase containing GFP was subjected to rotary evaporation of DIMCARB. The stability of GFP was not affected by the distillation of DIMCARB, and the DIMCARB was successfully recycled in three successive rounds of GFP purification. The potential of PPG + DIMCARB + water system as a sustainable protein purification tool is promising.
利用环境友好且可循环利用的离子液体形成双水相体系（ATPS）在蛋白质分离领域越来越受到关注。在本研究中，由N,N-二甲基氨基甲酸N,N-二甲基铵（DIMCARB）和热响应性聚丙二醇（PPG）组成的双水相体系被用于从大肠杆菌中回收重组绿色荧光蛋白（GFP）。通过改变PPG的分子量和双水相体系的总组成，系统地研究了GFP在PPG + DIMCARB + 水体系中的分配行为。总体而言，发现GFP优先分配到富含亲水性DIMCARB的相中。由42%（w/w）的PPG 1000和4.4%（w/w）的DIMCARB组成的双水相体系在GFP选择性（1237）和产率（98.8%）方面表现出最佳性能。该最佳体系还成功放大了50倍，且不影响纯化性能。含有GFP的下层相进行了DIMCARB的旋转蒸发。GFP的稳定性不受DIMCARB蒸馏的影响，并且DIMCARB在连续三轮的GFP纯化中成功循环使用。PPG + DIMCARB + 水体系作为一种可持续的蛋白质纯化工具具有很大的潜力。
