Biosensor H2O2 by Using Immobilized Horseradish Peroxidase Glutaraldehyde
An enzymatic biosensor has been developed for detection of hydrogen peroxide with immobilized Horseradish peroxidase (HRP). HRP was immobilized by using glutaraldehyde (GA) that cross linked with modified polyaniline (PANI) as mediator to improve electron transfer. Modified carbon paste electrodes (MCPE) PANI was more effective during electron transfer compared to carbon paste electrodes (CPE). Cyclic voltammetry method (VC) was used to determine the electrochemical properties of the modified electrode substrate to produce redox reactions. The effect of pH and temperature were analysed by cyclic voltammetry. The optimum performance of HRP/GA/PANI was at pH 7 and 50°C. Kinetics parameters HRP enzyme were determined in optimum condition. The Michaelis-Menten constant (Km) value and current maximum (Imax) have been obtained as 1.71 mM and 0.29 mA.
Hydrogen peroxide is one of the molecules involved in reactive oxygen species. Hydrogen peroxide are produced by catalysis of glucose oxidase, cholesterol oxidase, xanthine oxidase, alcohol oxidase, and uricase reaction. These enzymes were applied for monitoring levels of glucose, cholesterol, xanthine, alcohol, and gout. The advantage of using enzyme as component of biosensor is high selectivity to interact with specific substrates.
There are several enzymes that can be used for biosensors development such as catalase
- Cholesterol oxidase
- Glucose oxidase
- Bilirubin oxidase
- Laccase and others.
Horseradish peroxidase (HRP) is one of enzyme that used in electrochemistry as biosensor and biofuel cell, as decolorization agent, immunoassay, biodegradation and synthesis of polyaniline. The horseradish peroxidase (184.108.40.206) is classified as oxido reductase enzyme that can be used as biosensors. HRP enzyme can be isolated from some organisms such as roots of plant radish, E. coli, mammalian cells and yeast.
One of the critical point should be considered to make biosensor is electron transfer. Electron transfer is more efficient by using metal, but the price is more expensive and needed to replace metal. Other compound that can be used for electron transfer is polyaniline. Polyaniline has unique properties was called doping dedoping or protonation deprotonation.
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