Department of Biological Chemistry, Rothamsted Research, Harpenden, Herts, UK.
Plant Biotechnol J. 2012 Feb;10(2):195-206. doi: 10.1111/j.1467-7652.2011.00653.x. Epub 2011 Sep 8.
The role of acyl-CoA-dependent Δ6-desaturation in the heterologous synthesis of omega-3 long-chain polyunsaturated fatty acids was systematically evaluated in transgenic yeast and Arabidopsis thaliana. The acyl-CoA Δ6-desaturase from the picoalga Ostreococcus tauri and orthologous activities from mouse (Mus musculus) and salmon (Salmo salar) were shown to generate substantial levels of Δ6-desaturated acyl-CoAs, in contrast to the phospholipid-dependent Δ6-desaturases from higher plants that failed to modify this metabolic pool. Transgenic plants expressing the acyl-CoA Δ6-desaturases from either O. tauri or salmon, in conjunction with the two additional activities required for the synthesis of C20 polyunsaturated fatty acids, contained higher levels of eicosapentaenoic acid compared with plants expressing the borage phospholipid-dependent Δ6-desaturase. The use of acyl-CoA-dependent Δ6-desaturases almost completely abolished the accumulation of unwanted biosynthetic intermediates such as γ-linolenic acid in total seed lipids. Expression of acyl-CoA Δ6-desaturases resulted in increased distribution of long-chain polyunsaturated fatty acids in the polar lipids of transgenic plants, reflecting the larger substrate pool available for acylation by enzymes of the Kennedy pathway. Expression of the O. tauriΔ6-desaturase in transgenic Camelina sativa plants also resulted in the accumulation of high levels of Δ6-desaturated fatty acids. This study provides evidence for the efficacy of using acyl-CoA-dependent Δ6-desaturases in the efficient metabolic engineering of transgenic plants with high value traits such as the synthesis of omega-3 LC-PUFAs.
酰基辅酶 A 依赖性 Δ6-去饱和酶在异源合成 ω-3 长链多不饱和脂肪酸中的作用在转基因酵母和拟南芥中得到了系统评价。来自微微藻甲藻(Ostreococcus tauri)的酰基辅酶 A Δ6-去饱和酶和来自小鼠(Mus musculus)和三文鱼(Salmo salar)的同源活性被证明能够产生大量的 Δ6-去饱和酰基辅酶 A,与高等植物中依赖磷脂的 Δ6-去饱和酶形成对比,后者未能修饰这种代谢池。表达来自甲藻或三文鱼的酰基辅酶 A Δ6-去饱和酶的转基因植物,与合成 C20 多不饱和脂肪酸所需的另外两种活性一起,与表达琉璃苣磷脂依赖性 Δ6-去饱和酶的植物相比,含有更高水平的二十碳五烯酸。使用酰基辅酶 A 依赖性 Δ6-去饱和酶几乎完全消除了总种子脂质中不必要的生物合成中间产物如 γ-亚麻酸的积累。酰基辅酶 A Δ6-去饱和酶的表达导致转基因植物极性脂质中长链多不饱和脂肪酸的分布增加,反映出可用作 Kennedy 途径酶酰化的较大底物池。甲藻 Δ6-去饱和酶在转基因荠蓝植物中的表达也导致了 Δ6-去饱和脂肪酸的高水平积累。本研究为使用酰基辅酶 A 依赖性 Δ6-去饱和酶在高效代谢工程中构建具有高附加值性状的转基因植物(如 ω-3 LC-PUFA 的合成)提供了证据。