文献类型：期刊文献

英文题名：Separation of transglutaminase using aqueous two-phase systems composed of two pH-response polymers

作者：Dong, Lili[1];Wan, Junfen[1];Cao, Xuejun[1]

机构：[1]East China Univ Sci & Technol, Dept Bioengn, State Key Lab Bioreactor Engn, 130 Meilong Rd, Shanghai 200237, Peoples R China

年份：2018

卷号：1555

起止页码：106

外文期刊名：JOURNAL OF CHROMATOGRAPHY A

收录：;EI(收录号：20181905144470);WOS:【SCI-EXPANDED(收录号:WOS:000432762900012)】；

基金：The authors very appreciate financial support from the National Natural Science Foundation of China (No. 21376078).

语种：英文

外文关键词：Aqueous two-phase systems; pH-Responsive copolymer; Separation; Transglutaminase

摘要：Aqueous two-phase systems (ATPS) have been effectively used as a rapid and economical method for the separation and purification of many enzymes or proteins. However, a key problem is the recovery of the polymers forming ATPS and there are rarely available studies about ATPS for the transglutaminase. In this study, a pH-responsive ATPS has been established by two pH-responsive polymers (P-ADB4.91 and P-ADB4.06) that can be recycled by changing the pH values, with high recovery of over 96%. And partitioning of the crude transglutaminase in this new ATPS was investigated for the first time. The main parameters, such as crude TGase feedstock load, the pH of system (pH 6.50-7.80), polymers concentration, and the types and concentration of salts, were studied to optimize partition conditions. In the 3% P-ADB4.91/2% P-ADB4.06 ATPS, enzyme recovery of 96.51%, partition coefficient of 4.23 and purification factor of 3.73 for TGase were obtained in the presence of 60mmol/L MgSO4 and at pH 7.00. The result of sodium dodecyl sulfate-polyacrylamide gel electrophoresis shows that TGase can be well separated from crude extract. (C) 2018 Elsevier B.V. All rights reserved.