ACS Synthetic Biology
ACS Synthetic Biology
5 years ago

Co-Compartmentation of Terpene Biosynthesis and Storage via Synthetic Droplet

Co-Compartmentation of Terpene Biosynthesis and Storage via Synthetic Droplet
Man Li, Connor Gorman, Cheng Zhao, Susie Y. Dai, Cheng Hu, Xin Wang, Joshua S. Yuan, Yining Zeng, YongKyoung Kim, Shi-You Ding
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.

Publisher URL: http://dx.doi.org/10.1021/acssynbio.7b00368

DOI: 10.1021/acssynbio.7b00368

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