Improvement of arachidonic acid and eicosapentaenoic acid production by increasing the copy number of the genes encoding fatty acid desaturase and elongase into Pichia pastoris
2009
Li, Yun-Tao | Li, Mao-Teng | Fu, Chu-Hua | Zhou, Peng-Peng | Liu, Jian-Min | Yu, Long-Jiang
Genes encoding Δ6 desaturase, Δ6 fatty acid elongase, and Δ5 desaturase from the alga, Phaeodactylum tricornutum, were co-expressed in Pichia pastoris to produce arachidonic acid (ARA; 20:4 Δ⁵, ⁸, ¹¹, ¹⁴) and eicosapentaenoic acid (EPA; 20:5 Δ⁵, ⁸, ¹¹, ¹⁴, ¹⁷). A panel of Pichia clones carrying progressively increasing copies of the heterologous gene expression cassette was created using an in vitro multimerization approach. ARA and EPA accumulated up to 0.3 and 0.1% of total fatty acids, respectively, in the recombinant P. pastoris carrying with double copies of these three heterologous genes, as compared to 0.1 and 0.05%, respectively, in the recombinant P. pastoris with single copy.
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