Single Enzyme Nanoparticles in Nanoporous Silica: A Heirachical Approach to Enzyme Stabilization and Immobilization
Single enzyme nanoparticles of alpha-chymotrypsin (SEN-CT), in which each CT molecule is surrounded by a thin polymeric organic/inorganic network, stabilized the CT activity in a shaking condition as well as in a non-shaking condition. Since SEN-CT is soluble in a buffer solution and less than 10 nm in size, SEN-CT could be immobilized in nanoporous silica with an average pore size of 29 nm. Free CT and SEN-CT were immobilized in nanoporous silica (NPS), and nanoporous silica that was first silanized with aminopropyltriethoxysilane (amino-NPS) to generate a positive surface charge. The SEN-CT adsorbed in amino-NPS was more stable than CT immobilized by either adsorption in NPS or covalent bonding to amino-NPS. In shaking conditions, nanoporous silica provided an additional stabilization by protecting SEN-CT from shear stresses. At 22oC with harsh shaking, free, NPS- adsorbed and NPS-covalently-attached CT showed half lives of 1, 62, and 80 h, respectively; whereas SEN-CT adsorbed in amino-NPS showed no activity loss within 12 days. The combination of SENs and nanoporous silica, which makes an active and stable immobilized enzyme system, represents a new structure for biocatalytic applications.
- Research Organization:
- Pacific Northwest National Lab. (PNNL), Richland, WA (United States)
- Sponsoring Organization:
- USDOE
- DOE Contract Number:
- AC05-76RL01830
- OSTI ID:
- 883669
- Report Number(s):
- PNNL-SA-42088; EMTED2; KP1301020; TRN: US200614%%682
- Journal Information:
- Enzyme and Microbial Technology, 39:474-480, Vol. 39; ISSN 0141-0229
- Country of Publication:
- United States
- Language:
- English
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