A novel method for enzyme immobilization: direct encapsulation of acid phosphatase in nanoporous silica host materials.
Immobilization of acid phosphatase (ACP) in mesoporous or, more generally, nanoporous silica has been accomplished via the sol-gel reactions of tetramethyl orthosilicate in the presence of ACP and of D-glucose (DG) as a nonsurfactant template, which is subsequently removed by water extraction after the formation of nanocomposite gels. Characterization of the silica host after the removal of DG shows that the pore size and volume generally increase with the DG content. At high DG contents, the silica hosts are nanoporous with interconnected nanoscaled pores/channels of regular diameter (e.g., 3.4 nm). Catalytic activity of ACP encapsulated in nanoporous hosts is significantly improved over that in microporous host prepared in the absence of DG. The apparent enzymatic activity at various pH values and substrate concentrations correlates well with the nanostructures of the host matrices. As the DG content is increased in the synthesis, the activity tends to increase. At a DG content of 42-60 wt%, the samples exhibit activities about triple that of the template-free control. These and other results from enzymatic kinetic studies suggest that the increase in the pore size and volume facilitates the transport of the substrate and product molecules in the host matrices, leading to the observed increase in activity. The thermal stability of ACP is remarkably improved upon immobilization. There is no detectable leakage of ACP from the host matrices and the biogels are reuseable. This study provides a useful protocol for the development of nanotechnology for various biocatalysts and biosensors.
Wei
Y
Y
Department of Chemistry, Drexel University, Philadelphia, Pennsylvania 19104, USA.
Xu
J
J
Feng
Q
Q
Lin
M
M
Dong
H
H
Zhang
W J
WJ
Wang
C
C
D000135:Acid Phosphatase / Q000138:chemical synthesis / Q000737:chemistry*; D000327:Adsorption; D004339:Drug Compounding / Q000379:methods*; D004789:Enzyme Activation; D004795:Enzyme Stability; D004800:Enzymes, Immobilized / Q000138:chemical synthesis / Q000737:chemistry*; D005947:Glucose / Q000737:chemistry*; D006863:Hydrogen-Ion Concentration; D008854:Microscopy, Electron; D008968:Molecular Conformation; D036103:Nanotechnology / Q000379:methods*; D016062:Porosity; D058428:Silica Gel; D012822:Silicon Dioxide / Q000737:chemistry*; D013379:Substrate Specificity; D013696:Temperature
D004800:Enzymes, Immobilized; D058428:Silica Gel; D012822:Silicon Dioxide; D000135:Acid Phosphatase; D005947:Glucose