Co-Compartmentation of Terpene Biosynthesis and Storage via Synthetic Droplet,ACS Synthetic Biology

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Co-Compartmentation of Terpene Biosynthesis and Storage via Synthetic Droplet
ACS Synthetic Biology ( IF 3.7 ) Pub Date : 2018-02-13 00:00:00 , DOI: 10.1021/acssynbio.7b00368
Cheng Zhao _{1,

2,

3} , YongKyoung Kim _{1,

2,

3} , Yining Zeng ₄ , Man Li _{1,

2,

3} , Xin Wang _{1,

2,

3} , Cheng Hu _{1,

2,

3} , Connor Gorman _{1,

2,

3} , Susie Y Dai ₅ , Shi-You Ding ₆ , Joshua S Yuan _{1,

2,

3}

Affiliation

Traditional bioproduct engineering focuses on pathway optimization, yet is often complicated by product inhibition, downstream consumption, and the toxicity of certain products. Here, we present the co-compartmentation of biosynthesis and storage via a synthetic droplet as an effective new strategy to improve the bioproduct yield, with squalene as a model compound. A hydrophobic protein was designed and introduced into the tobacco chloroplast to generate a synthetic droplet for terpene storage. Simultaneously, squalene biosynthesis enzymes were introduced to chloroplasts together with the droplet-forming protein to co-compartmentalize the biosynthesis and storage of squalene. The strategy has enabled a record yield of squalene at 2.6 mg/g fresh weight without compromising plant growth. Confocal fluorescent microscopy imaging, stimulated Raman scattering microscopy, and droplet composition analysis confirmed the formation of synthetic storage droplet in chloroplast. The co-compartmentation of synthetic storage droplet with a targeted metabolic pathway engineering represents a new strategy for enhancing bioproduct yield.

中文翻译：

通过合成液滴进行萜烯生物合成和储存的共区室

传统的生物产品工程侧重于途径优化，但往往因产品抑制、下游消耗和某些产品的毒性而变得复杂。在这里，我们提出了通过合成液滴进行生物合成和储存的共同区室，作为提高生物产品产量的有效新策略，并以角鲨烯作为模型化合物。设计了一种疏水蛋白并将其引入烟草叶绿体中，以产生用于萜烯储存的合成液滴。同时，将角鲨烯生物合成酶与液滴形成蛋白一起引入叶绿体，以共同划分角鲨烯的生物合成和储存。该策略在不影响植物生长的情况下，实现了创纪录的角鲨烯产量，达到 2.6 毫克/克鲜重。共焦荧光显微镜成像、受激拉曼散射显微镜和液滴成分分析证实了叶绿体中合成存储液滴的形成。合成存储液滴与目标代谢途径工程的共同划分代表了提高生物产品产量的新策略。

更新日期：2018-02-13

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