US 11,884,954 B2
Protein complex based on DNA enzymes of E family of Escherichia coli and application thereof in artificial protein scaffolds
Lixin Ma, Wuhan (CN); Chao Zhai, Wuhan (CN); Jun Yang, Wuhan (CN); and Fei Wang, Wuhan (CN)
Assigned to Hubel University, Wuhan (CN)
Filed by Hubei University, Wuhan (CN)
Filed on Dec. 29, 2022, as Appl. No. 18/148,284.
Application 18/148,284 is a continuation of application No. PCT/CN2022/102567, filed on Jun. 30, 2022.
Claims priority of application No. 202111256498.4 (CN), filed on Oct. 27, 2021.
Prior Publication US 2023/0220439 A1, Jul. 13, 2023
Int. Cl. C12N 9/22 (2006.01); C12N 9/14 (2006.01); C12N 15/52 (2006.01); C07K 14/245 (2006.01); C12P 21/06 (2006.01); C12N 15/62 (2006.01); C12P 21/02 (2006.01)
CPC C12P 21/06 (2013.01) [C07K 14/245 (2013.01); C12N 15/62 (2013.01); C12P 21/02 (2013.01)] 3 Claims
 
1. A method for preparing a multienzyme complex system, comprising:
connecting a carbohydrate-binding module 3a (CBM3a), an immunity 2 (Im2) protein, an Im7 protein, an Im8 protein, and an Im9 protein in series through linkers to obtain a scaffold protein;
forming interaction pairs by connecting a CL2 protein, a CL7 protein, a CL8 protein and a CL9 protein respectively with the Im2 protein, the Im7 protein, the Im8 protein, and the Im9 protein on the scaffold protein;
connecting target proteins to the CL2 protein, the CL7 protein, the CL8 protein and the CL9 protein respectively to realize an orderly assembly of the target proteins on the scaffold protein, so as to obtain the multienzyme complex system;
wherein the amino acid sequence of the CL2 protein is shown in SEQ ID NO: 2;
wherein the amino acid sequence of the CL7 protein is shown in SEQ ID NO: 5;
wherein the amino acid sequence of the CL8 protein is shown in SEQ ID NO: 8;
wherein the amino acid sequence of the CL9 protein is shown in SEQ ID NO: 11;
wherein the amino acid sequence of the Im2 protein is shown in SEQ ID NO: 3;
wherein the amino acid sequence of the Im7 protein is shown in SEQ ID NO: 6;
wherein the amino acid sequence of the Im8 protein is shown in SEQ ID NO: 9; and
wherein the amino acid sequence of the Im9 protein is shown in SEQ ID NO: 12.