Difference between revisions of "Part:BBa K4891023"
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Phosphoenolpyruvate (PEP) and erythrose-4-phosphate (E4P) could be stereospecifically condensed into 3-deoxy-D-arabino-heptulosonate-7-phosphate (DAHP) by tktA/talB gene encoding DAHP synthase. | Phosphoenolpyruvate (PEP) and erythrose-4-phosphate (E4P) could be stereospecifically condensed into 3-deoxy-D-arabino-heptulosonate-7-phosphate (DAHP) by tktA/talB gene encoding DAHP synthase. | ||
| − | <!-- Add more about the biology of this part here | + | <!-- Add more about the biology of this part here--> |
===Usage and Biology=== | ===Usage and Biology=== | ||
| + | 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. See table below: | ||
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| + | | <html><img style="width:400px" src="https://static.igem.wiki/teams/4891/wiki/parts/22.png" ></html> | ||
| + | |- | ||
| + | |} | ||
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<span class='h3bb'>Sequence and Features</span> | <span class='h3bb'>Sequence and Features</span> | ||
Latest revision as of 15:16, 9 October 2023
tktA
Phosphoenolpyruvate (PEP) and erythrose-4-phosphate (E4P) could be stereospecifically condensed into 3-deoxy-D-arabino-heptulosonate-7-phosphate (DAHP) by tktA/talB gene encoding DAHP synthase.
Usage and Biology
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. See table below:
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Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 232
Illegal AgeI site found at 337
Illegal AgeI site found at 1669 - 1000COMPATIBLE WITH RFC[1000]