Evolved sortases and uses thereof
AssigneePresident and Fellows of Harvard College
InventorsDavid R. Liu, Brent M. Dorr
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Abstract
Evolved sortases exhibiting enhanced reaction kinetics and/or altered substrate preferences are provided herein, for example evolved sortases that bind recognitions motifs comprising a LAXT or LPXS sequence. Also provided are methods (e.g., orthogonal transpeptidation and diagnostics methods) for using such sortases. Kits comprising materials, reagents, and cells for carrying out the methods described herein are also provided.
Claims28
A Staphylococcus aureus (S. aureus) Sortase A variant that binds substrates comprising the sequence LAXT, wherein X represents any amino acid, and wherein the S. aureus Sortase A variant comprises an amino acid sequence that is at least 90% identical to the amino acid sequence of S. aureus Sortase A as provided as SEQ ID NO:
- 1, and wherein the amino acid sequence of the Sortase A variant includes a mutation selected from the group consisting of K84R, R99H, R99K, S102C, A104H, E105D, K138I, K138V, K138P, K145E, K152I, D160K, K162R, K162H, T164N, V168I, K177G, K177R, I182F, and K196S.
A Staphylococcus aureus (S. aureus) Sortase A variant that binds substrates comprising the sequence LPXS, wherein X represents any amino acid, the S. aureus Sortase A variant comprising an amino acid sequence that is at least 90% identical to the amino acid sequence of S. aureus Sortase A as provided as SEQ ID NO:
- 1, wherein the amino acid sequence of the Sortase A variant includes a mutation selected from the group consisting of N98D, S102C, A104V, A118S, A118T, F122A, K134G, K134P, E189V, E189F, and E189P.
Dependencies are inferred from the claim text.