Development of cholesterol biosensor with high sensitivity using dual-enzyme immobilization into the mesoporous silica materials |
| |
Authors: | Kazuki Murai Katsuya Kato |
| |
Institution: | 1. Department of Applied Chemistry, Graduate School of Engineering, Chubu University, 1200 Matsumoto-cho, Kasugai-si, 487-8501, Japan;2. Bio-Integrated Processing Group, Advanced Manufacturing Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), 2266-98, Anagahora, Shimosidami, Moriyama-ku, Nagoya, 463-8510, Japan |
| |
Abstract: | Mesoporous silica (MPS) materials with different pore diameters were synthesized by a sol–gel method where organic templates such as cationic surfactant (cetyltrimethylammonium bromide) and triblock co-polymer of (poly(ethylene glycol)–poly(propylene glycol)–poly(ethylene glycol) (Pluronic P123, EO20PO70EO20)), were used. MPS surface was organo-functionalized using a silane coupling reagent (ethyl-, phenyl-, or 3-mercaptpropyltriethoxysilane). Dual-enzyme, cholesterol esterase (10.0 nm × 5.4 nm × 11.0 nm) and cholesterol oxidase (6.8 nm × 8.5 nm × 8.8 nm), was immobilized on MPS materials by physical adsorption. Amount of dual-enzyme immobilized on all MPS materials, having a different pore size (2.7, 6.4, 12.4, 14.7, and 22.6 nm), and organo-functionalized MPS was similar (CE: 1.5 mg/mg silica and CO: 0.01 mg/mg silica). High activity of dual-enzyme was obtained by adjacently immobilizing on MPS materials. Its activity on MPS-2 (pore diameter: 6.4 nm) or MPS-5 (pore diameter: 22.6 nm) showed approximately 60% of native activity. Moreover, dual-enzyme immobilized on MPS with highly hydrophobic organo-functional groups (phenyl- or mercaptopropyl-group) exhibited higher activity than that on no-substituted MPS. Relative activity of dual-enzyme immobilized on organo-functionalized MPS-2 increased from 58% to 93%, under the optimum conditions. |
| |
Keywords: | |
本文献已被 ScienceDirect 等数据库收录! |
|