Coupling metabolic addiction with negative autoregulation to improve strain stability and pathway yield
| dc.contributor.author | Lv, Yongkun | |
| dc.contributor.author | Gu, Yang | |
| dc.contributor.author | Xu, Jingliang | |
| dc.contributor.author | Zhou, Jingwen | |
| dc.contributor.author | Xu, Peng | |
| dc.date.accessioned | 2020-08-17T17:32:24Z | |
| dc.date.available | 2020-08-17T17:32:24Z | |
| dc.date.issued | 2020-05-20 | |
| dc.description.abstract | Metabolic addiction, an organism that is metabolically addicted with a compound to maintain its growth fitness, is an underexplored area in metabolic engineering. Microbes with heavily engineered pathways or genetic circuits tend to experience metabolic burden leading to degenerated or abortive production phenotype during long-term cultivation or scale-up. A promising solution to combat metabolic instability is to tie up the end-product with an intermediary metabolite that is essential to the growth of the producing host. Here we present a simple strategy to improve both metabolic stability and pathway yield by coupling chemical addiction with negative autoregulatory genetic circuits. Naringenin and lipids compete for the same precursor malonyl-CoA with inversed pathway yield in oleaginous yeast. Negative autoregulation of the lipogenic pathways, enabled by CRISPRi and fatty acid-inducible promoters, repartitions malonyl-CoA to favor flavonoid synthesis and increased naringenin production by 74.8%. With flavonoid-sensing transcriptional activator FdeR and yeast hybrid promoters to control leucine synthesis and cell grwoth fitness, this amino acid feedforward metabolic circuit confers a flavonoid addiction phenotype that selectively enrich the naringenin-producing pupulation in the leucine auxotrophic yeast. The engineered yeast persisted 90.9% of naringenin titer up to 324 generations. Cells without flavonoid addiction regained growth fitness but lost 94.5% of the naringenin titer after cell passage beyond 300 generations. Metabolic addiction and negative autoregulation may be generalized as basic tools to eliminate metabolic heterogeneity, improve strain stability and pathway yield in long-term and large-scale bioproduction. | en_US |
| dc.description.sponsorship | This work was supported by the Cellular & Biochem EngineeringProgram of the National Science Foundation under grant no.1805139and the Bill and Melinda Gates Foundation (OPP1188443). YL wouldlike to thank the China Scholarship Council, China Postdoctoral ScienceFoundation (Grant No. 2019M662535), and Science and TechnologyProject of Henan Province (Grant No. 202102310019) for funding support. | en_US |
| dc.description.uri | https://www.sciencedirect.com/science/article/pii/S1096717620300938?via%3Dihub | en_US |
| dc.format.extent | 10 pages | en_US |
| dc.genre | journal articles | en_US |
| dc.identifier | doi:10.13016/m2kkt2-edj9 | |
| dc.identifier.citation | Yongkun Lv et al., Coupling metabolic addiction with negative autoregulation to improve strain stability and pathway yield, Metabolic Engineering Volume 61, Pages 79-88 (2020), https://doi.org/10.1016/j.ymben.2020.05.005 | en_US |
| dc.identifier.uri | https://doi.org/10.1016/j.ymben.2020.05.005 | |
| dc.identifier.uri | http://hdl.handle.net/11603/19443 | |
| dc.language.iso | en_US | en_US |
| dc.publisher | Elsevier | en_US |
| dc.relation.isAvailableAt | The University of Maryland, Baltimore County (UMBC) | |
| dc.relation.ispartof | UMBC Chemical, Biochemical & Environmental Engineering Department Collection | |
| dc.relation.ispartof | UMBC Faculty Collection | |
| dc.rights | This item is likely protected under Title 17 of the U.S. Copyright Law. Unless on a Creative Commons license, for uses protected by Copyright Law, contact the copyright holder or the author. | |
| dc.rights | Attribution 4.0 International | * |
| dc.rights.uri | https://creativecommons.org/licenses/by/4.0/ | * |
| dc.title | Coupling metabolic addiction with negative autoregulation to improve strain stability and pathway yield | en_US |
| dc.type | Text | en_US |