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Conflict of Interest: In relation to this article, we declare that there is no conflict of interest.

Publication history: Received August 23, 2023
Accepted August 23, 2023

This is an Open-Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/bync/3.0) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Covalent crowding strategy for trypsin confined in accessible mesopores with enhanced catalytic property and stability

Cheng Zhou^{1 2} Bo Jiang^{1 2} Zecui Sheng^{1 2} Shemin Zhu^{1 3} Shubao Shen^{1 2†}

¹National Engineering Research Center for Biotechnology, Nanjing 210009, P. R. China ²College of Biotechnology and Pharmaceutical Engineering, Nanjing University of Technology, Nanjing 210009, P. R. China ³College of Material Science and Engineering, Nanjing University of Technology, Nanjing 210009, P. R. China

zsbshen@gmail.com

Korean Journal of Chemical Engineering, March 2011, 28(3), 853-859(7), 10.1007/s11814-010-0412-3

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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_x000D_ thermal stability of BSA-modified preparation at 50℃ 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 ℃. 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.

Keywords

Immobilized Trypsin Covalent Modification Crowding Environment Accessible Transmission

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