Development of a catalase based biosensor for alcohol determination in beer samples

Area of Science:

• Biotechnology
• Electrochemistry
• Enzyme Technology

Background:

• Alcohol determination is crucial in various industries, including food and beverage.
• Existing methods for alcohol analysis can be time-consuming or require complex instrumentation.
• Development of rapid, sensitive, and cost-effective biosensors is highly desirable.

Purpose of the Study:

• To develop and characterize an amperometric biosensor for alcohol determination using catalase enzyme.
• To investigate the immobilization strategy and optimize the working conditions for the biosensor.
• To compare the performance of the developed biosensor with a standard enzymatic-spectrophotometric method.

Main Methods:

• Immobilization of catalase enzyme onto a Clark-type dissolved oxygen probe using gelatin and glutaraldehyde.
• Utilizing the oxygen-producing reaction of catalase with hydrogen peroxide and its subsequent consumption in the presence of ethanol.
• Amperometric detection of changes in dissolved oxygen concentration to quantify ethanol.

Main Results:

• The biosensor demonstrated a linear response to ethanol concentration in the range of 0.05 to 1.0 mM.
• A low detection limit of 0.05 mM and a rapid response time of 3 minutes were achieved.
• Optimal working conditions were determined as pH 7.0 (50 mM phosphate buffer) and 35°C.
• The biosensor exhibited good reproducibility, substrate specificity, and operational/storage stability.
• Results obtained from alcoholic drinks using the biosensor correlated well with the enzymatic-spectrophotometric method.

Conclusions:

• The developed catalase-based amperometric biosensor is a viable tool for accurate and efficient alcohol determination.
• The biosensor offers advantages in terms of speed, sensitivity, and simplicity compared to traditional methods.
• This technology holds potential for application in quality control and analysis of alcoholic beverages.