We engineered the shikimate pathway of E. coli MG1655 to efficiently accumulate SA. We knocked out ptsG, ldhA, adhE, poxB, and pta genes to achieve the production of precursor substance PEP. To increase intracellular E4P content, we overexpressed tktA and talB genes. To enhance product accumulation, we overexpressed aroG, aroB, aroD, and aroE genes, while knocking down aroK and aroL genes to cut off the metabolic flux, thus accomplishing the accumulation of shikimic acid. Besides, a non-phosphorylated pathway, that is, glk and glf genes (mainly by glk-glf integration into the ptsG locus) is introduced to enhance glucose utilization. To achieve the goal above-mentioned, we totally constructed 26 parts this year.
No. | part Number | Type | part Name |
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Enhance shikimic acid synthesis metabolic flux |
1 | BBa_K4891000 | Basic | aroGfbr |
2 | BBa_K4891001 | Basic | aroB |
3 | BBa_K4891002 | Basic | aroD |
4 | BBa_K1218008 | Basic | aroE |
5 | BBa_K4891005 | Composite | aroGfbr-aroB-aroD-aroE |
Increase the E4P content |
6 | BBa_K4891023 | Basic | tktA |
7 | BBa_K4891024 | Basic | talB |
8 | BBa_K4891025 | Composite | tktA-talB |
9 | BBa_K4891022 | Basic | RBS |
Blocking the PTS and Enhancing glucose utilization |
10 | BBa_K4891003 | Basic | glk |
11 | BBa_K4891004 | Basic | glf |
12 | BBa_K4891016 | Basic | ptsGup |
13 | BBa_K4891017 | Basic | ptsGdown |
14 | BBa_K4891014 | Composite | glk-glf |
15 | BBa_K4891015 | Composite | ptsGup-glk-glf-ptsGdown |
Block the shikimic acid catabolic pathway |
16 | BBa_K4891006 | Basic | aroK |
17 | BBa_K4891007 | Basic | aroL |
18 | BBa_K4891010 | Basic | sgRNAaroK |
19 | BBa_K4891011 | Basic | sgRNAaroL |
Blocking PEP branching pathways |
20 | BBa_K4891008 | Basic | ldhA |
21 | BBa_K4891009 | Basic | adhE |
22 | BBa_K4891012 | Basic | poxB |
23 | BBa_K4891013 | Basic | pta |
24 | BBa_K4891018 | Basic | sgRNAldhA |
25 | BBa_K4891019 | Basic | sgRNAadhE |
26 | BBa_K4891020 | Basic | sgRNApoxB |
27 | BBa_K4891021 | Basic | sgRNApta |