Enhancing isoprenoid synthesis in Yarrowia lipolytica by expressing the isopentenol utilization pathway and modulating intracellular hydrophobicity
Enhancing isoprenoid synthesis in Yarrowia lipolytica by expressing the isopentenol utilization pathway and modulating intracellular hydrophobicity
复制标题
通过表达异戊烯醇利用途径和调节细胞内疏水性增强解脂耶氏酵母中的类异戊二烯合成
DOI：
10.1016/j.ymben.2020.07.010
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发表时间：
2020-09-01
影响因子：
8.4
通讯作者：
Stephanopoulos, Gregory
中科院分区：
文献类型：
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作者：
Luo, Zhengshan;Liu, Nian;Stephanopoulos, Gregory
生物合成前体的丰富供应和产物与细胞内环境的相容性对微生物类异戊二烯的生产起着重要作用。在这项研究中，我们量身定制这两个要求，通过引入两步异戊烯醇利用途径（IUP），以增加在产油酵母解脂耶氏酵母的本地途径。与普通的类异戊二烯前体异戊烯基焦磷酸（IPP）及其异构体二甲基烯丙基焦磷酸（DMAPP）的快捷方式相比，IUP能够将IPP + DMAPP水平提高15.7倍。IPP + DMAPP水平的增加可以直接导致更好的类异戊二烯合成，这一点使用番茄红素作为模型化合物进行了说明。此外，我们还证明了细胞中较高的脂质含量与改善的细胞内番茄红素生产相关，这表明具有大量疏水环境以螯合类异戊二烯的重要性。将这些策略与进一步的遗传和发酵优化相结合，我们实现了4.2 g/ L的最终番茄红素滴度。总的来说，这些策略在加强微生物中长链类异戊二烯和脂溶性天然产物的合成方面具有很大的潜力。
The abundant supply of biosynthetic precursors and product compatibility with the intracellular environment play important roles for microbial isoprenoid production. In this study, we tailor to both of these requirements by introducing the two-step isopentenol utilization pathway (IUP) to augment the native pathway in the oleaginous yeast Yarrowia lipolytica. With shortcut access to the common isoprenoid precursor, isopentenyl pyrophosphate (IPP) and its isomer dimethylallyl pyrophosphate (DMAPP), IUP is capable of elevating IPP + DMAPP levels by 15.7-fold compared to the mevalonate pathway alone. The increase in IPP + DMAPP levels can directly lead to better isoprenoid synthesis, which is illustrated using lycopene as a model compound. Moreover, we also demonstrate that higher lipid contents in the cells correlate with improved intracellular lycopene production, suggesting the importance of having a substantial hydrophobic environment to sequester isoprenoids. Combining these strategies with further genetic and fermentation optimizations, we achieved a final lycopene titer of 4.2 g/ L. Overall, these strategies hold great potential for strengthening the synthesis of long-chain isoprenoids and fatsoluble natural products in microbes.