Background Yarrowia lipolytica, a non-traditional oil yeast, has been widely used as a platform for lipid production. However, the production of other chemicals such as terpenoids in engineered Y. lipolytica is still low. α-Farnesene, a sesquiterpene, can be used in medicine, bioenergy and other fields, and has very high economic value. Here, we used α-farnesene as an example to explore the potential of Y. lipolytica for terpenoid production. Results We constructed libraries of strains overexpressing mevalonate pathway and α-farnesene synthase genes by non-homologous end-joining (NHEJ) mediated integration into the Y. lipolytica chromosome. First, a mevalonate overproduction strain was selected by overexpressing relevant genes and changing the cofactor specificity. Based on this strain, the downstream α-farnesene synthesis pathway was overexpressed by iterative integration. Culture conditions were also optimized. A strain that produced 25.55 g/L α-farnesene was obtained. This is the highest terpenoid titer reported in Y. lipolytica. Conclusions Yarrowia lipolytica is a potentially valuable species for terpenoid production, and NHEJ-mediated modular integration is effective for expression library construction and screening of high-producer strains.

中文翻译：

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工程化产油酵母解脂耶氏酵母以生产 α-法呢烯。

背景解脂耶氏酵母是一种非传统的油酵母，已被广泛用作脂质生产的平台。然而，工程化解脂耶氏酵母中萜类化合物等其他化学物质的产量仍然很低。α-法呢烯是一种倍半萜烯，可用于医药、生物能源等领域，具有很高的经济价值。在这里，我们以 α-法呢烯为例来探索解脂耶氏酵母生产萜类化合物的潜力。结果我们通过非同源末端连接（NHEJ）介导的整合到解脂耶氏酵母染色体中构建了过表达甲羟戊酸途径和α-法呢烯合酶基因的菌株文库。首先，通过过度表达相关基因和改变辅因子特异性来选择甲羟戊酸过度生产菌株。基于这种应变，下游α-法呢烯合成途径通过迭代整合过表达。培养条件也进行了优化。获得了产生25.55 g/L α-法呢烯的菌株。这是在解脂耶氏酵母中报道的最高的萜类化合物滴度。结论 解脂耶氏酵母是一种潜在的萜类生产有价值的物种，NHEJ介导的模块化整合对于表达文库的构建和高产菌株的筛选是有效的。