Combinatorial Modulation of Linalool Synthase and Farnesyl Diphosphate Synthase for Linalool Overproduction in Saccharomyces cerevisiae,Journal of Agricultural and Food Chemistry

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Combinatorial Modulation of Linalool Synthase and Farnesyl Diphosphate Synthase for Linalool Overproduction in Saccharomyces cerevisiae
Journal of Agricultural and Food Chemistry ( IF 5.7 ) Pub Date : 2021-01-11 , DOI: 10.1021/acs.jafc.0c06384
Pingping Zhou _{1,

2} , Yi Du _{1,

2} , Xin Fang _{1,

2} , Nannan Xu ₂ , Chunlei Yue ₂ , Lidan Ye ₃

Affiliation

Linalool, as a fragrant monoterpene, is an important feedstock for food, pharmaceuticals, and cosmetics industries. Although our previous study had significantly increased linalool production by the directed evolution of linalool synthase and overexpression of the whole mevalonate pathway genes, the engineered yeast strain suffered from dramatically reduced biomass. Herein, a stress-free linalool-producing yeast cell factory was constructed by the combinational regulation of linalool synthase and farnesyl diphosphate synthase instead of multienzyme overexpression. First, the expression level of linalool synthase was successfully enhanced by introducing a N-terminal SKIK tag, which improved linalool production by 3.3-fold. Subsequently, the modular assembly of linalool synthase and dominant negative farnesyl diphosphate synthase via short peptide tags efficiently converted geranyl pyrophosphate to linalool. Additional downregulation of the native farnesyl diphosphate synthase led to the highest reported linalool production (80.9 mg/L) in yeast. This combinatorial modulation strategy may also be applied to the production of other high-value monoterpenes.

中文翻译：

酿酒酵母中芳樟醇过量生产的芳樟醇合酶和法呢基二磷酸合酶的组合调制。

芳樟醇是一种芳香的单萜，是食品，制药和化妆品行业的重要原料。尽管我们先前的研究通过芳樟醇合酶的定向进化和整个甲羟戊酸途径基因的过表达而显着增加了芳樟醇的产量，但工程酵母菌株的生物量却大大减少。在此，通过组合调节芳樟醇合酶和法呢基二磷酸合酶而不是多酶过表达来构建无胁迫的芳樟醇生产酵母细胞工厂。首先，通过引入N末端SKIK标签成功提高了芳樟醇合成酶的表达水平，从而将芳樟醇的产量提高了3.3倍。后来，经由短肽标签的芳樟醇合酶和显性负法呢基二磷酸合磷酸酶合成模块组装有效地将香叶基焦磷酸转化为芳樟醇。天然法呢基二磷酸合酶的进一步下调导致酵母中芳樟醇的产量最高（80.9 mg / L）。该组合调制策略也可以应用于其他高价值单萜的生产。

更新日期：2021-01-27

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