In general, promoters have significant influence on recombinant protein production. Herein, we compared the performance of actin (pACT), phosphoglycerate kinase (pPGK), and translational elongation factor (pTEF) promoters for driving the expression of fatty acid delta-6 (Δ6) desaturase from Nile tilapia (Oreochromis niloticus; Oni-fads2) in Saccharomyces cerevisiae. Our results showed that by applying real-time RT-PCR, the highest level of Oni-fads2 mRNA was observed in S. cerevisiae carrying the expression vector driven by pTEF promoters. Exogenous substrate C18:2n-6 was used to determine Δ6 activity by quantitatively determining the C18:3n-6 product. The results showed that highest Δ6 desaturation was observed when using pTEF as a promoter. Recombinant S. cerevisiae cells expressing Oni-fads2 driven by pTEF were tested with the substrate C18:3n-3, and Δ6 desaturation efficiently converted C18:3n-3 to C18:4n-3. Furthermore, crude extract of recombinant yeast also exhibited Δ6 activity. Thus, recombinant S. cerevisiae cells expressing Oni-fads2 driven by the pTEF promoter have potential as a yeast factory for the sustainable production of long-chain polyunsaturated fatty acids.

中文翻译：

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无处不在的启动子指导酿酒酵母中尼罗罗非鱼（Oreochromis niloticus）的脂肪酸Delta-6去饱和酶的表达。

通常，启动子对重组蛋白的产生有重要影响。在这里，我们比较了肌动蛋白（pACT），磷酸甘油酸激酶（pPGK）和翻译延伸因子（pTEF）启动子从尼罗罗非鱼（Oreochromis niloticus; Oni-fads2）驱动脂肪酸delta-6（Δ6）去饱和酶表达的性能。 ）酿酒酵母中。我们的结果表明，通过实时RT-PCR，在携带由pTEF启动子驱动的表达载体的酿酒酵母中观察到了最高水平的Oni-fads2 mRNA。通过定量确定C18：3n-6产物，使用外源底物C18：2n-6确定Δ6活性。结果显示当使用pTEF作为启动子时观察到最高的Δ6去饱和。表达由pTEF驱动的Oni-fads2的酿酒酵母细胞用底物C18进行了测试：3n-3和Δ6去饱和将C18：3n-3有效地转换为C18：4n-3。此外，重组酵母的粗提物也表现出Δ6活性。因此，表达由pTEF启动子驱动的Oni-fads2的重组酿酒酵母细胞具有作为酵母工厂可持续生产长链多不饱和脂肪酸的潜力。