EVOLVED SORTASES AND USE THEREOF

摘要

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.

主权项

1. A sortase that binds substrates comprising the sequence LAXT, wherein X represents any amino acid, and wherein the sortase 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, or a fragment thereof, wherein the amino acid sequence of the sortase includes one or more mutations selected from the group consisting of K84R, R99H or R99K, S102C, A104H, E105D, K138I or K138V or K138P, K145E, K152I, D160K, K162R or K162H, T164N, V168I, K177G or K177R, I182F, or T196S.