公开(公告)号：US20240376160A1

公开(公告)日：2024-11-14

申请号：US18255368

申请日：2022-08-30

Applicant: NANJING TECH UNIVERSITY

Inventor： Ling JIANG ,  Yao CHEN ,  Yujin CHEN ,  Liying ZHU ,  Wei LIU

IPC: C07K14/315 ,  C12N15/70 ,  C12R1/19

Abstract: The invention relates to a molecular peptide mutant with a high ester bond formation efficiency, the amino acid sequence of which is as shown in SEQ ID NO: 1. The invention introduces mutations at three sites on the basis of not affecting the spatial structure of Catcher, and through three mutations, obtains the Catcher mutant EBCatcher, which significantly improves the stability of the Catcher in water solution and can improve the binding efficiency to EBTag.

公开(公告)号：US20240002454A1

公开(公告)日：2024-01-04

申请号：US18255108

申请日：2022-04-25

Applicant: NANJING TECH UNIVERSITY

Inventor： Ling JIANG ,  Yao CHEN ,  Liying ZHU ,  Wei LIU ,  Xianhan CHEN ,  Xinyi CHEN

IPC: C07K14/315 ,  C12N15/70

CPC classification number: C07K14/315 ,  C12N15/70

Abstract: The invention relates to a molecular peptide mutant, the amino acid sequence of which is as shown in SEQ ID NO: 1. In the invention, SpyCatcher is designed and modified to obtain a molecular peptide SpyCatcher-21 with stimulus response to pH on the basis of not affecting the formation of isopeptide bonds, and Pro is introduced into a key loop of the SpyCatcher-21 through analysis of the crystal structure to reduce the flexibility of the loop and obtain a mutant SpyCatcher-21_A82P, which can raise ligation efficiency with SpyTag. The SpyCatcher-21_A82P can be used to achieve double-enzyme catalysis according to objective needs by changing the pH of the environment to obtain different degrees of coupling, or obtain a three-enzyme coupled catalytic system through electrostatic interaction with a positively charged enzyme.