<br>用于 8-羟基香叶醇高效生物合成的工程酵母底盘的构建和优化,mLife

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用于 8-羟基香叶醇高效生物合成的工程酵母底盘的构建和优化
mLife Pub Date : 2023-12-26 , DOI: 10.1002/mlf2.12099
Yu Zhang _{1,

2,

3} , Mengdi Yuan ₁ , Xinxin Wu ₁ , Qiuhui Zhang ₁ , Yuzhu Wang ₁ , Liming Zheng ₁ , Tsan-Yu Chiu ₁ , Huiming Zhang ₁ , Lei Lan ₁ , Feng Wang ₁ , Ying Liao ₁ , Xuemei Gong _{1,

3} , Shirui Yan _{1,

3,

4} , Yun Wang _{1,

3,

4} , Yue Shen _{1,

3,

4} , Xian Fu _{1,

3,

4}

Affiliation

单萜类吲哚生物碱（MIA）的微生物生产为获得具有高药用价值的化合物提供了一种可持续且环保的方法。然而，由于由香叶醇 8-羟化酶（G8H）催化的香叶醇及其衍生物 8-羟基香叶醇等关键前体的供应不足，异源微生物中 MIA 的高效生物合成受到阻碍。在这项研究中，我们开发了一个简单的进化平台，通过使用嵌入 Sc2.0 合成酵母中的 SCRaMbLE 系统来筛选香叶醇产量提高的菌株，并证实了相关基因组靶标的因果作用。通过基因组挖掘，我们确定了几种 G8H 酶，它们在体内生产 8-羟基香叶醇方面的表现比常用的 CrG8H 好得多。我们进一步表明，通过交换实验，这些 G8H 酶的 N 端在细胞活性中起重要作用。最后，工程底盘、优化的生物合成途径和 G8H 的利用相结合，导致最终菌株的产生 8-羟基香叶醇比起始菌株提高了 30 倍以上。总体而言，本研究将为酵母细胞的构建和优化提供见解，以实现 8-羟基香叶醇及其衍生物的高效生物合成。

"点击查看英文标题和摘要"

The construction and optimization of engineered yeast chassis for efficient biosynthesis of 8-hydroxygeraniol

Microbial production of monoterpenoid indole alkaloids (MIAs) provides a sustainable and eco-friendly means to obtain compounds with high pharmaceutical values. However, efficient biosynthesis of MIAs in heterologous microorganisms is hindered due to low supply of key precursors such as geraniol and its derivative 8-hydroxygeraniol catalyzed by geraniol 8-hydroxylase (G8H). In this study, we developed a facile evolution platform to screen strains with improved yield of geraniol by using the SCRaMbLE system embedded in the Sc2.0 synthetic yeast and confirmed the causal role of relevant genomic targets. Through genome mining, we identified several G8H enzymes that perform much better than the commonly used CrG8H for 8-hydroxygeraniol production in vivo. We further showed that the N-terminus of these G8H enzymes plays an important role in cellular activity by swapping experiments. Finally, the combination of the engineered chassis, optimized biosynthesis pathway, and utilization of G8H led to the final strain with more than 30-fold improvement in producing 8-hydroxygeraniol compared with the starting strain. Overall, this study will provide insights into the construction and optimization of yeast cells for efficient biosynthesis of 8-hydroxygeraniol and its derivatives.

更新日期：2023-12-26

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