METHOD FOR DETERMINING ACTIVITY OF SORTASE A Russian patent published in 2025 - IPC C12N1/20 C12R1/445 

FIELD: biotechnology.

SUBSTANCE: invention relates to biotechnology, in particular to a method for determining activity of sortase A, comprising adding to a cell culture of S. aureus isolated and purified protein GFP-LPGTG with an additional sequence on the C-end of the LPGTG protein, which is recognized by sortase A, after which sortase A covalently attaches the protein with said amino acid residues to the cell wall of S. aureus and S. aureus cells begin to fluoresce in the range of GFP protein, if sortase is active. 5 ml of said mixture is separated and used as a negative control of inhibitory activity against grade A, and the other two portions of 5 ml of the mixture are added with LPRDA inhibitor, or KUD748, or KUD146. All portions of the mixture are incubated, cells are washed from GFP-LPGTG protein, resuspended, and fluorescence of S. aureus cell suspension treated with LPRDA or KUD748, or KUD146 inhibitor is measured. Fluorescence of control and S. aureus cell suspension treated with LPRDA or KUD748 or KUD146 inhibitor is compared.

EFFECT: invention provides more effective testing of antimicrobial substances acting on sortase, and obtaining a definite answer on the effect of the tested substance on sortase A of Staphylococcus aureus.

1 cl, 5 dwg, 4 ex