An easily regenerable enzyme reactor prepared from polymerized high internal phase emulsions

作者：Ruan, Guihua^*; Wu, Zhenwei; Huang, Yipeng; Wei, Meiping; Su, Rihui; Du, Fuyou^*

来源：Biochemical and Biophysical Research Communications, 2016, 473(1): 54-60.

DOI：10.1016/j.bbrc.2016.03.049

• Immobilized enzymatic reactor
• Protein digestion
• High internal phase emulsion
• PolyHIPE monoliths

A large-scale high-efficient enzyme reactor based on polymerized high internal phase emulsion monolith (polyHIPE) was prepared. First, a porous cross -linked polyHIPE monolith was prepared by in-situ thermal polymerization of a high internal phase emulsion containing styrene, divinylbenzene and polyglutaraldehyde. The enzyme of TPCK-Trypsin was then immobilized on the monolithic polyHIPE. The performance of the resultant enzyme reactor was assessed according to the conversion ability of Nebenzoyl-L-arginine ethyl ester to 1%-benzoyl-r-arginine, and the protein digestibility of bovine serum albumin (BSA) and cytochrome (Cyt-C). The results showed that the prepared enzyme reactor exhibited high enzyme immobilization efficiency and fast and easy -control protein digestibility. BSA and Cyt-C could be digested in 10 min with sequence coverage of 59% and 78%, respectively. The peptides and residual protein could be easily rinsed out from reactor and the reactor could be regenerated easily with 4 M HCl without any structure destruction. Properties of multiple interconnected chambers with good permeability, fast digestion facility and easily reproducibility indicated that the polyHIPE enzyme reactor was a good selector potentially applied in proteomics and catalysis areas.

• 出版日期2016-4-22
• 单位桂林理工大学