Difference between revisions of "Part:BBa K4343076"

Latest revision as of 12:58, 10 October 2022

pUC-HUH-PaDes17

The PaDes17 gene encodes a ∆-17 elongase from Pythium aphanidermatum that catalyzes the conversion of arachidonic acid (ARA; C20:4 n-6) to Eicosapentaenoic acid (EPA), an ω 3 polyunsaturated fatty acid (PUFA).

Characterisation

Two copies of this gene and two copies of ∆-5 desaturase (EgDes5) From Euglena gracilis were randomly integrated into the genome of the Engineered strain Po1F-19 by transformation. The ∆-17 desaturase (PaDes17) gene was expressed under the action of promoter TEF to generate ∆-17 elongase, which can catalyze Arachidonic acid (ARA; C20:4n-6) generated Eicosapentaenoic acid (EPA), the engineered strain Po1F-20 was obtained. After fermentation and culture, the effect of the increase of the copy number of ∆-17 elongase and ∆-5 desaturase on the increase of EPA content was tested by gas chromatography.

Result

Po1f-20 was further randomly integrated with 2 copies of EgDes5 and 2 copies of PaDes17 in the genome of the engineered strain Po1f-19, and EPA proportion reached 22.4% of the TFAs. (Fig: Po1f-20 )

Sequence and Features