期刊
JOURNAL OF MATERIALS CHEMISTRY
卷 21, 期 43, 页码 17468-17475出版社
ROYAL SOC CHEMISTRY
DOI: 10.1039/c1jm11598g
关键词
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资金
- National Institute of Environmental Health Sciences [U54 ES16015]
- National Institute of Health (NIH)
- NIH through the National Institute of Neurological Disorders and Stroke [U01 NS058161-01]
- U.S. Department of Energy (DOE) [DE-AC05-76RL01830]
- Department of Energy's Office of Biological and Environmental Research
- National Natural Science Foundation of China [20975037]
Apoferritin is a uniform spherical nano-size biomaterial with excellent bio-compatibility. In this work, we report the use of apoferritin as a novel biocompatible carrier for stabilizing enzymes and enhancing their activities. We used glucose oxidase (GOx) as a model enzyme in this study. GOx was immobilized on the surface of the apoferritin through a green synthetic approach, taking advantage of bioaffinity binding between streptavidin and biotin. As a result, a glucose oxidase-biotin/streptavidin/biotin-apoferritin conjugate (Apo-GOx) was prepared using streptavidin as the bridge. The synthesized Apo-GOx was characterized by transmission electron microscopy, ultraviolet and fluorescence spectroscopy. The activity and stability of GOx on the surface of the apoferritin were investigated and challenged by different environmental factors, such as the temperature, chemicals and pH, in comparison with the biotinylated GOx (B-GOx). The results demonstrate that the activity of Apo-GOx is significantly enhanced while the thermal and chemical stabilities of Apo-GOx are also greatly improved compared to free B-GOx. For instance, the activity of the Apo-GOx only lost 30% after 2 h incubation at 50 oC in comparison to a 70% loss of free B-GOx. The activity of Apo-GOx remains intact after 30 min incubation in 5 M urea solution while B-GOx lost 80% activity after the same treatment. Furthermore, glucose detection was used as a model application for the enzyme immobilization method developed in this work. The GOx immobilized apoferritin nanoparticles exhibited high sensitivity for glucose detection with a detection limit of 3 nM glucose. This work offers a novel approach for immobilizing enzymes with enhanced stability and activity, thus holds the promising advantage for a number of applications, such as in enzyme catalysis, DNA assays and immunoassays.
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