Engineered ethanol-driven biosynthetic system for improving production of acetyl-CoA derived drugs in Crabtree-negative yeast
| Autor: | Qi Liu, Mingqiang Xu, Xiangshan Zhou, Menghao Cai, Zhilan Qian, Yuanxing Zhang, Jiahui Yu, Chenxiao Bai, Yiqi Liu, Qin Xu, Qiangqiang Peng |
|---|---|
| Rok vydání: | 2019 |
| Předmět: |
0106 biological sciences
Bioengineering Naphthalenes 01 natural sciences Applied Microbiology and Biotechnology Metabolic engineering 03 medical and health sciences chemistry.chemical_compound Biosynthesis Acetyl Coenzyme A Yeasts 010608 biotechnology Transcriptional regulation Monacolin J 030304 developmental biology 0303 health sciences Ethanol Acetyl-CoA Metabolism Yeast Biosynthetic Pathways Cytosol Metabolic Engineering chemistry Biochemistry Biotechnology |
| Zdroj: | Metabolic Engineering. 54:275-284 |
| ISSN: | 1096-7176 |
| DOI: | 10.1016/j.ymben.2019.05.001 |
| Popis: | Many natural drugs use acetyl-CoA as the key biosynthetic precursor. While in eukaryotic chassis host like yeast, efficient biosynthesis of these drugs is often hampered by insufficient acetyl-CoA supply because of its compartmentalized metabolism. Reported acetyl-CoA engineering commonly modifies central carbon metabolism to pull and push acetyl-CoA into cytosol from sugars or redirects biosynthetic pathways in organelles, involving complicated metabolic engineering strategies. We constructed a new biosynthetic system based on a Crabtree-negative yeast, which grew exceptionally on ethanol and assimilated ethanol directly in cytosol to acetyl-CoA (3 steps). A glucose-repressed and ethanol-induced transcriptional signal amplification device (ESAD) with 20-fold signal increase was constructed by rewiring native transcriptional regulation circuits. This made ethanol the sole and fast-growing substrate, acetyl-CoA precursor, and strong biosynthetic pathway inducer simultaneously. The ESAD was used for biosynthesis of a commercial hypolipidemic drug intermediate, monacolin J. A strain producing dihydromonacolin L was firstly constructed and systematically engineered. We further developed a coculture system equipped with this upstream strain and a downstream strain with dihydromonacolin L–to–monacolin J module controlled by a synthetic constitutive transcriptional signal amplification device (CSAD). It produced a high monacolin J titre of 2.2 g/L on ethanol in bioreactor. Engineering glucose-supported and ethanol-repressed fatty acids biosynthesis in the upstream strain contributed more acetyl-CoA for monacolin J and improved its titre to 3.2 g/L, far surpassing other reported productions in yeasts. This study provides a new paradigm for facilitating the high-yield production of acetyl-CoA derived pharmaceuticals and value-added molecules. |
| Databáze: | OpenAIRE |
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