METHOD FOR MONITORING ACTIVITY OF SINGLE ENZYME MOLECULES Russian patent published in 2024 - IPC C12M1/00 C12M1/34 C12Q1/25 

FIELD: nanotechnology.

SUBSTANCE: invention relates to enzymology of single molecules and can be used to study enzymatic activities of single enzyme molecules using nanotechnology, as well as processes of denaturation of single enzyme molecules. Method for monitoring the activity of single molecules of the enzyme of the class of oxidoreductases by the example of horseradish peroxidase and the class of hydrolases by the example of asparaginase is carried out using a nanopore detector. Nanopore detector comprises a cuvette with cis- and trans-chambers, between which there is a nano-pore in the form of a cone made by the electron-beamdrilling (EBD) method in Six-Ny. Method involves introducing a buffer solution containing an enzyme into a cis chamber of the cuvette, and a buffer solution without an enzyme into the trans chamber. Voltage is supplied between cis- and trans-chambers, with the help of which a single enzyme molecule is introduced into the nanopore with its subsequent immobilization. Immobilisation process is controlled by the change in the electric current flowing through the nanopore. Cis chamber is washed from non-immobilized enzyme molecules. Buffer solution of a substrate inducing enzyme activity is added to the cis-chamber. Change in the current flowing through the nanopore is used to monitor the activity of the enzyme during interaction of its molecule with the molecule of the introduced buffer solution of the substrate.

EFFECT: group of inventions provides a wider range of technical means for realizing the method of monitoring activity of single molecules.

4 cl, 3 dwg, 3 ex