Details

Autor(en) / Beteiligte

• Ruan, Guihua
• Wu, Zhenwei
• Huang, Yipeng
• Wei, Meiping
• Su, Rihui
• Du, Fuyou

Titel

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

Ist Teil von

• Biochemical and biophysical research communications, 2016-04, Vol.473 (1), p.54-60

Ort / Verlag

United States: Elsevier Inc

Erscheinungsjahr

2016

Quelle

MEDLINE

Beschreibungen/Notizen

• 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 Nα-benzoyl-l-arginine ethyl ester to Nα-benzoyl-l-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. Schematic illustration of preparation of hypercrosslinking polyHIPE immobilized enzyme reactor for on-column protein digestion. [Display omitted] •A reactor was prepared and used for enzyme immobilization and continuous on-column protein digestion.•The new polyHIPE IMER was quite suit for protein digestion with good properties.•On-column digestion revealed that the IMER was easy regenerated by HCl without any structure destruction.