Part:BBa_K5235010
GMAS-PPK-BsAld-CsAlaDC-GNP1
In 2024, SHSBNU-China aims to use synthetic biology techniques to genetically edit the E. coli strain BL21 to produce L-theanine. We designed a plasmid containing five genes (BsAld, CsAlaDC, GMAS, PPK, and GNP1) responsible for the production and transport of theanine on the common vector pET28a. The plasmid is designed to be expression in E. coli with IPTG controlled induction, followed by HP-LC testing of the supernatant to determine the yield.
We used five genes in total: BsAld, CsAlaDC, GMAS, PPK, and GNP1. - BsAld, CsAlaDC, and GMAS are responsible for the de novo synthesis of theanine, starting from glucose. BsAld and CsAlaDC carry out the conversion of pyruvate to ethylamine, and GMAS synthesizes theanine from ethylamine and glutamate, which is produced by the generic bacterial TCA cycle. - PPK provides ATP to aid the synthesis pathway. - GNP1 is responsible for transporting the synthesized theanine out of the bacterial cells.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
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