Covalent crowding strategy for trypsin confined in accessible mesopores with enhanced catalytic property and stability |
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Authors: | Cheng Zhou Bo Jiang Zecui Sheng Shemin Zhu Shubao Shen |
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Affiliation: | (1) National Engineering Research Center for Biotechnology, College of Life Science and Pharmaceutical Engineering, Nanjing University of Technology, No.5 Xinmofan Road, Nanjing, 210009, People’s Republic of China;(2) College of Materials Science and Engineering, Nanjing University of Technology, Nanjing, 210009, People’s Republic of China; |
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Abstract: | Chemically modified macromolecules were assembled with adsorptive trypsin in mesoporous silica foams (MCFs) to establish covalent
linkage. Effects of catalytic properties and stability of immobilized trypsin were examined. The addition of chemically modified
protein (BSA) and polysaccharide (ficoll) to the immobilized trypsin exhibited high coupled yield (above 90%) and relative
activities (174.5% and 175.9%, respectively), showing no protein leaching after incubating for 10 h in buffers. They showed
broader pH and temperature profiles, while the half life of thermal stability of BSA-modified preparation at 50 °C increased
to 1.3 and 2.3 times of unmodified and free trypsin, respectively. The modified trypsin in aqueous-organic solvents exhibited
100% activity after 6 h at 50 °C. The kinetic parameters of trypsin preparations and suitable pore diameter of MCFs warranted
compatibility of covalent modification for substrate transmission. The covalent crowding modification for immobilized trypsin
in nanopores establishes suitable and accessible microenvironment and renders possibility of biological application. |
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