학술논문

Improved Method for Immobilization of Trypsin to Enhance Catalytic Performance of Trypsin in Confined Nanospaces
Document Type
Journal Article
Source
JOURNAL OF CHEMICAL ENGINEERING OF JAPAN. 2010, 43(7):595
Subject
Immobilization
Macromolecular Crowding
Mesocellular Siliceous Foams
Trypsin
Language
English
ISSN
0021-9592
Abstract
Mesocellular siliceous foams (MCFs) functionalized with different groups were used in the trypsin immobilization process. Macromolecular reagents were co-assembled with trypsin in the MCFs to create a microenvironment resembling that in living cells. The catalytic characteristics and stability of trypsin preparations were investigated. The relative activity of immobilized trypsin was 122.9% when the enzyme was immobilized adsorptively in the MCFs. Trypsin assembled with BSA in the MCFs attained maximum specific activity, 1.33 times that of the unmodified immobilized trypsin. The Km value of the BSA derivative of trypsin was reduced to 15.9% and 16.6% relative to the valve for native and solely immobilized trypsin, respectively, while the Kcat/Km value of trypsin assembled with BSA was enhanced to 148%. The residual activity of trypsin assembled with BSA was 48.5% and 29.4% at 50°C and 60°C, respectively, after incubation for 60 min.