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Delta 5 Fatty Acid Desaturase Upregulates the Synthesis of Polyunsaturated Fatty Acids in the Marine Diatom Phaeodactylum tricornutum

机译:Delta 5脂肪酸去饱和酶上调海洋硅藻Phaeodactylum tricornutum中多不饱和脂肪酸的合成

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摘要

Microalgae are important primary producers in the marine ecosystem and excellent sources of lipids and other bioactive compounds. The marine diatom Phaeodactylum tricornutum accumulates eicosapentaenoic acid (EPA, 20:5n-3) as its major component of fatty acids. To improve the EPA production, delta 5 desaturase, which plays a role in EPA biosynthetic pathway, was characterized in P. tricornutum. An annotated delta 5 desaturase PtDSb gene was cloned and overexpressed in P. tricornutum. The transgene was integrated into the genome demonstrated by Southern blot, and the overexpression of PtDSb was verified by qPCR and Western blot analysis. Fatty acid composition exhibited a significant increase in the unsaturated fatty acids. Monounsaturated fatty acids (MUFA) and polyunsaturated fatty acids (PUFA) showed an increase of 75% and 64%, respectively. In particular, EPA showed an increase of 58% in engineered microalgae. Meanwhile, neutral lipid content showed an increase up to 65% in engineered microalgae. More importantly, engineered cells showed a similar growth rate with the wild type, thus keeping high biomass productivity. This work provides an effective way to improve the production of microalgal value-added bioproducts by metabolic engineering.
机译:微藻是海洋生态系统中重要的主要生产者,也是脂质和其他生物活性化合物的绝佳来源。海洋硅藻Phaeodactylum tricornutum积累二十碳五烯酸(EPA,20:5n-3)作为其脂肪酸的主要成分。为了提高EPA的产量,在三角果假单胞菌中鉴定了在EPA生物合成途径中起作用的delta 5去饱和酶。一个带注释的delta 5去饱和酶PtDSb基因被克隆并在三角果假单胞菌中过表达。通过Southern印迹证实转基因已整合到基因组中,并且通过qPCR和Western印迹分析证实了PtDSb的过表达。脂肪酸组成显示出不饱和脂肪酸的显着增加。单不饱和脂肪酸(MUFA)和多不饱和脂肪酸(PUFA)分别增加了75%和64%。特别是,EPA显示工程微藻的含量增加了58%。同时,工程微藻中的中性脂质含量增加了高达65%。更重要的是,工程细胞显示出与野生型相似的生长速率,从而保持了高生物量生产率。这项工作为通过代谢工程提高微藻增值生物产品的生产提供了有效途径。

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