期刊
MATERIALS SCIENCE & ENGINEERING C-MATERIALS FOR BIOLOGICAL APPLICATIONS
卷 33, 期 3, 页码 1438-1445出版社
ELSEVIER SCIENCE BV
DOI: 10.1016/j.msec.2012.12.048
关键词
Titania; Covalent binding; Immobilized enzyme; Biocatalysis; Stability; Kinetic parameters
资金
- National Science Foundation of China [21076145]
- Natural Science Foundation of Tianjin [09JCYBJC06700]
- Program for New Century Excellent Talents in University [NCET-10-0623]
- National Basic Research Program of China [2009CB724705]
- National Science Fund for Distinguished Young Scholars [21125627]
- Program of Introducing Talents of Discipline to Universities [B06006]
- Open Funding Project of the State Key Laboratory of Bioreactor Engineering
In this study, a novel approach combing the chelation and covalent binding was explored for facile and efficient enzyme immobilization. The unique capability of titania to chelate with catecholic derivatives at ambient conditions was utilized for titania surface functionalization. The functionalized titania was then used for enzyme immobilization. Titania submicrospheres (500-600 nm) were synthesized by a modified sol-gel method and functionalized with carboxylic acid groups through a facile chelation method by using 3-(3,4-dihydroxyphenyl) propionic acid as the chelating agent. Then, catalase (CAT) was covalently immobilized on these functionalized titania submicrospheres through 1-ethyl-3-[3-dimethylaminopropyl] carbodiimide hydrochloride/N-hydroxysuccinimide (EDC/NHS) coupling reaction. The immobilized CAT retained 65% of its free form activity with a loading capacity of 100-150 mg/g titania. The pH stability, thermostability, recycling stability and storage stability of the immobilized CAT were evaluated. A remarkable enhancement in enzyme stability was achieved. The immobilized CAT retained 90% and 76% of its initial activity after 10 and 16 successive cycles of decomposition of hydrogen peroxide, respectively. Both the K-m and the V-max values of the immobilized CAT (27.4 mM, 13.36 mM/min) were close to those of the free CAT (25.7 mM, 13.46 mM/min). (C) 2012 Elsevier B.V. All rights reserved.
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