Immobilized mitochondrial electron transport particle for NADH determination

Area of Science:

• Biochemistry
• Electrochemistry
• Biotechnology

Background:

• Electron transport particles (ETP) from mitochondria are crucial for cellular respiration.
• Developing reusable biosensors enhances analytical efficiency and sustainability.
• Enzyme immobilization is a key technique in biosensor development.

Purpose of the Study:

• To immobilize nonphosphorylating electron transport particles (ETP) from beef heart mitochondria in agar gel.
• To develop a reusable electrochemical biosensor for determining NADH and succinate.
• To assess the specificity of the biosensor using malonate inhibition.

Main Methods:

• Preparation and immobilization of beef heart ETP in agar gel.
• Assembly of an electrochemical device utilizing immobilized ETP and an oxygen probe.
• Measurement of oxidase activity towards NADH and succinate.
• Evaluation of succinate detection specificity through malonate inhibition.

Main Results:

• Immobilized ETP exhibited oxidase activity for both NADH and succinate.
• The immobilized ETP demonstrated reusability in the biosensor.
• The electrochemical device successfully detected NADH and succinate.
• Malonate addition specifically inhibited the biosensor's response to succinate.

Conclusions:

• Immobilized beef heart ETP can be effectively used in a reusable electrochemical biosensor.
• The developed biosensor allows for the determination of NADH and succinate.
• The biosensor exhibits specificity for succinate detection, as shown by malonate inhibition.