Manufacturing Multienzymatic Complex Reactors In Vivo by Self-Assembly To Improve the Biosynthesis of Itaconic Acid in Escherichia coli,ACS Synthetic Biology

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Manufacturing Multienzymatic Complex Reactors In Vivo by Self-Assembly To Improve the Biosynthesis of Itaconic Acid in Escherichia coli
ACS Synthetic Biology ( IF 3.7 ) Pub Date : 2018-05-04 00:00:00 , DOI: 10.1021/acssynbio.8b00086
Zhongwei Yang ₁ , Hongling Wang ₁ , Yuxiao Wang ₁ , Yuhong Ren ₁ , Dongzhi Wei ₁

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The self-assembly of multienzyme into bioreactors is of extensive interest to spatially regulate valuable reactions. Despite the important progresses achieved, methods to precisely manufacture multienzymatic complex reactors (MECRs) are still poorly proposed both in vivo and in vitro, particularly for more than three biocatalytically relevant enzymes. Here, we developed a sequential self-assembly system to form multitude MECRs involving three enzymes in the itaconic acid (IA) pathway with two pairs of protein–peptide interactions. The MECRs were identified as nanoscale particle-like structures when self-assembled in vitro and produced higher IA production than the unassembled and linearly assembled systems when applied in vivo coupling with CRISPR-Cas9 based metabolic engineering. This work provides novel insights into the construction of multifarious multienzyme complex into bioreactors by the self-assembly strategy for multistep cascades to sequentially control metabolic fluxes inside cells.

中文翻译：

通过自组装体内制造多酶复合反应器以改善大肠杆菌中衣康酸的生物合成

将多酶自组装到生物反应器中对于在空间上调节有价值的反应具有广泛的兴趣。尽管取得了重要的进展，但在体内和体外，尤其是对于三种以上生物催化相关的酶，仍很少有人提出精确制造多酶复合反应器（MECR）的方法。在这里，我们开发了一种顺序自组装系统，以形成衣康酸（IA）途径中涉及三种酶并具有两对蛋白质-肽相互作用的多种MECR。当在体外自组装时，将MECRs鉴定为纳米级颗粒状结构，并且在体内应用时，与未组装和线性组装的系统相比，产生的IA产量更高与基于CRISPR-Cas9的代谢工程结合。通过多步级联的自组装策略顺序控制细胞内的代谢通量，这项工作为将多种多样的多酶复合物构建到生物反应器中提供了新颖的见解。

更新日期：2018-05-04

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