Parts

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
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