화학공학소재연구정보센터
홈 로그인 로그아웃 연락처 사이트맵
Korean Journal of Chemical Engineering, Vol.28, No.3, 853-859, March, 2011
DOI10.1007/s11814-010-0412-3  Export Citation
Covalent crowding strategy for trypsin confined in accessible mesopores with enhanced catalytic property and stability
Cheng Zhou1, 2, Bo Jiang1, 2, Zecui Sheng1, 2, Shemin Zhu1, 3, and Shubao Shen1, 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
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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℃ 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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