Chemical & Biomolecular Engineering, Clemson University, Clemson, SC, USA.
Chemical & Environmental Engineering, University of California Riverside, Riverside, CA, USA.
Biotechnol J. 2017 Oct;12(10). doi: 10.1002/biot.201700248. Epub 2017 Sep 12.
The engineering of Yarrowia lipolytica to accumulate lipids with high titers and productivities has been enabled with a handful of constitutive promoters for pathway engineering. However, the development of promoters that are both strong and lipid responsive could greatly benefit the bioproduction efficiency of lipid-derived oleochemicals in oleaginous yeast. In this study, a fatty acid regulated hybrid promoter for use in Y. lipolytica is engineered. A 200 bp upstream regulatory sequence in the peroxisomal acyl CoA oxidase 2 (POX2) promoter is identified. Further analysis of the promoter sequence reveal a regulatory sequence, that when used in tandem repeats, lead to a 48-fold induction of gene expression relative to glucose and fourfold higher than the native POX2 promoter. To date, this is the strongest inducible promoter reported in Y. lipolytica. Taken together, the results show that it is possible to engineer strong promoters that retain strong inducibility. These types of promoters will be useful in controlling metabolism and as fatty acid sensors.
通过使用少数组成型启动子进行途径工程，已经能够使解脂耶氏酵母（Yarrowia lipolytica）积累高浓度和高产量的脂质。然而，开发既强又对脂质有响应的启动子将极大地提高油脂酵母中脂质衍生的油脂化学品的生物生产效率。在本研究中，设计了一种用于解脂耶氏酵母的脂肪酸调控杂交启动子。在过氧化物酶体酰基辅酶 A 氧化酶 2（POX2）启动子中鉴定了一个 200bp 的上游调控序列。对启动子序列的进一步分析表明，存在一个调控序列，当串联重复使用时，相对于葡萄糖诱导基因表达的倍数增加了 48 倍，比天然的 POX2 启动子高 4 倍。迄今为止，这是在解脂耶氏酵母中报道的最强诱导启动子。总之，这些结果表明，有可能设计出既强又保持强诱导性的启动子。这些类型的启动子将在控制代谢和作为脂肪酸传感器方面很有用。
