Difference between revisions of "Part:BBa K4343070"

Latest revision as of 11:47, 9 October 2022

TsElo5

The TsElo5 gene encodes a ∆-5 elongase from Thraustochytrium sp. that catalyzes the conversion of Eicosapentaenoic acid (EPA) to Docosapentaenoic acid (DPA), an ω-3 polyunsaturated fatty acid (PUFA).

Characterisation

We constructed a plasmid containing promoter TEF, TsELO5, and terminator Cyc1t through the T4 ligase. The plasmid was linearized by enzyme cleavage and then transformed into engineered strain Po1f-20 (22.4% of EPA in the TFAs) and randomly integrated into its genome. The TsElo5 gene was expressed in the presence of the promoter TEF, which can catalyze the conversion of Eicosapentaenoic acid (EPA; C20:5n-3) to Docosapentaenoic acid (DPA; C22:5n-3), and subsequently, the activity of TsElo5 was examined by gas chromatography.

Result

In order to obtain an engineered strain for co-production of DHA and EPA production, Po1f-20 was used as the starting strain. Besides, ∆-5 elongase were identified to convert EPA to DHA, so TsElo5 gene from Thraustochytrium sp. was randomly integrated in the genome of Po1f-20 to produce Docosapentaenoic acid (DPA; C22:5n-3). The GC analysis indicated that the percentage of EPA and DPA in the engineered strain were 3.5% and 23.9% of TFAs, respectively.

Sequence and Features