Determination of phospholipase D activity with a choline biosensor

Add time:07/23/2019    Source:sciencedirect.com

A choline biosensor for the determination of the phospholipase D activity in rape seeds was prepared. Choline oxidase was co-immobilized with catalase on a partially hydrolysed nylon net using glutaraldehyde and cyclohexyl isocyanide. The nylon net with immobilized enzymes was fixed on the tip of a Clark-type oxygen sensor. The hydrogen peroxide formed was removed by catalase to prevent inactivation of the choline oxidase. The prepared choline biosensor was characterized by the determination of the specific activity of immobilized choline oxidase (3.24 IU mg−1 at pH 8.0), pH and temperature effects on the enzyme activity, the apparent Michaelis constant (1.02 mM at pH 8.0) and the range of the linear biosensor response to the choline concentration (3.34 X 10−3 to 0.167 mM). The long-term biosensor ability is shown by its use for 600 assays over an 18-month period under different pH conditions. The determination of phospholipase D activity was performed in parallel with radiochemical assay and the results obtained demonstrated the priority of the biosensor application.

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