US 12,221,640 B2
Method for promoting n-acetylglucosamine synthesis by using GlcN6P responsive element
Long Liu, Wuxi (CN); Jian Chen, Wuxi (CN); Guocheng Du, Wuxi (CN); Yaokang Wu, Wuxi (CN); Taichi Chen, Wuxi (CN); and Jianghua Li, Wuxi (CN)
Assigned to JIANGNAN UNIVERSITY, Jiangsu (CN)
Appl. No. 17/279,271
Filed by JIANGNAN UNIVERSITY, Wuxi (CN)
PCT Filed Nov. 26, 2019, PCT No. PCT/CN2019/120949
§ 371(c)(1), (2) Date Mar. 24, 2021,
PCT Pub. No. WO2021/102682, PCT Pub. Date Jun. 3, 2021.
Prior Publication US 2022/0049280 A1, Feb. 17, 2022
Int. Cl. C12P 19/26 (2006.01); C12N 9/10 (2006.01); C12N 9/22 (2006.01); C12N 15/67 (2006.01); C12N 15/75 (2006.01)
CPC C12P 19/26 (2013.01) [C12N 9/1029 (2013.01); C12N 15/67 (2013.01); C12N 15/75 (2013.01); C12Y 203/01004 (2013.01); C12N 9/22 (2013.01)] 13 Claims
 
1. A method for promoting N-acetylglucosamine synthesis, comprising controlling the expression of glucosamine 6-phosphate N-acetyltransferase GNA4 by GNA1 by using a GlcN6P responsive element to dynamically regulate the N-acetylglucosamine synthesis pathway; and using the GlcN6P responsive element to regulate a compound formed by binding a dCas9 protein to three sgRNA expression fragments acting on zwf, pfkA and glmM genes, to dynamically weaken the glycolysis pathway, the pentose phosphate pathway and the peptidoglycan synthesis pathway, wherein the GlcN6P responsive element comprises the transcription factor GamR and a promoter containing a GamR binding site, where the transcription factor GamR has an amino acid sequence comprising positions 1-235 of an amino acid sequence deposited under NCBI Accession No.: WP_015382651.1, and the promoter is a PgamA promoter or a hybrid promoter constructed by adding a GamR binding site to a constitutive promoter.