<br>高效生物合成8-羟基香叶醇的工程酵母底盘的构建和优化,mLife

当前位置： X-MOL 学术 › › 论文详情

Our official English website, www.x-mol.net, welcomes your feedback! (Note: you will need to create a separate account there.)

高效生物合成8-羟基香叶醇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，由于香叶醇及其由香叶醇8-羟化酶（G8H）催化的衍生物8-羟基香叶醇等关键前体供应不足，异源微生物中MIA的有效生物合成受到阻碍。在本研究中，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，我们鉴定了几种 G8H 酶，其在体内生产 8-羟基香叶醇方面的表现比常用的 CrG8H 好得多。我们通过交换实验进一步表明这些 G8H 酶的 N 末端在细胞活性中发挥着重要作用。最后，工程底盘、优化的生物合成途径和 G8H 的利用相结合，最终菌株的 8-羟基香叶醇产量比起始菌株提高了 30 倍以上。总的来说，这项研究将为酵母细胞的构建和优化提供见解，以有效生物合成 8-羟基香叶醇及其衍生物。

"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

点击分享查看原文

点击收藏

公开下载