Journal
JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
Volume 67, Issue 35, Pages 9851-9857Publisher
AMER CHEMICAL SOC
DOI: 10.1021/acs.jafc.9b03727
Keywords
arachidonic acid; Yarrowia lipolytica; pathway engineering; protein engineering
Funding
- National Science Fund for Excellent Young Scholars of China [21922806]
- National Natural Science Foundation of China [21808052, 21776131, 21476111]
- Science and Technology Department of Hunan Province [2018RS3086]
- Scientific Research Fund of Hunan Provincial Education Department [18B090]
- Double first-class construction project of Hunan Agricultural University [SYL201802002]
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Arachidonic acid (ARA, C20:4) is a typical co-6 polyunsaturated fatty acid with special functions. Using Yarrowia lipolytica as an unconventional chassis, we previously showed the performance of the Delta-6 pathway in ARA production. However, a significant increase in the Delta-9 pathway has rarely been reported. Herein, the Delta-9 pathway from Isochrysis galbana was constructed via pathway engineering, allowing us to synthesize ARA at 91.5 mg L-1. To further improve the ARA titer, novel enzyme fusions of Delta-9 elongase and Delta-8 desaturase were redesigned in special combinations containing different linkers. Finally, with the integrated pathway engineering and synthetic enzyme fusion, a 29% increase in the ARA titer, up to 118.1 mg/L, was achieved using the reconstructed strain RH-4 that harbors the rigid linker (GGGGS). The results show that the combined pathway and protein engineering can significantly facilitate applications of Y. lipolytica.
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