Simple approach for efficient encapsulation of enzyme in silica matrix with retained bioactivity

We developed an alcohol-free sol-gel approach to encapsulate biomolecules such as horseradish peroxidase (HRP) in an electrochemically induced three-dimensional porous silica matrix by a one-step process. In this sol-gel process, the electrochemically generated hydroxyl ions at the electrode surface by applying cathodic current promote the hydrolysis of ammonium fluorosilicate to produce silica, and simultaneously the generated hydrogen bubbles play an important role in forming porous silica matrix. If HRP is mixed with ammonium fluorosilicate solution, it can be encapsulated in the forming silica matrix. Since there is no ethanol involved in the entire procedure, bioactivities of the encapsulated HRP can be effectively retained. As revealed by scanning electron microscopy (SEM) characterization, the resultant silica matrix has interconnected and network-like porous structures. Macroporous holes induced by hydrogen bubbles scattering on the relatively flat areas of porous structure can be observed. Such structure free from cracks provides effective mass transport and long-term stability. Scanning electrochemical microscope (SECM) characterization shows that the immobilized HRP molecules uniformly distribute in the silica matrix. The present HRP electrochemical biosensor exhibits a quick response (within 5 s) to H(2)O(2) in the concentration range from 0.02 to 0.20 mM (correlation coefficient of 0.9934) with a detection limit of 3 microM. The apparent Michaelis-Menten constant is 0.88 mM. The present alcohol-free sol-gel approach is effective for biomolecule encapsulation and is promising for the construction of biosensors, bioelectronics, and biofuel cells.