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Cell Rep ; 36(3): 109413, 2021 07 20.
Artículo en Inglés | MEDLINE | ID: mdl-34289355

RESUMEN

Metabolic regulation strategies have been developed to redirect metabolic fluxes to production pathways. However, it is difficult to screen out target genes that, when repressed, improve yield without affecting cell growth. Here, we report a strategy using a quorum-sensing system to control small RNA transcription, allowing cell-density-dependent repression of target genes. This strategy is shown with convenient operation, dynamic repression, and availability for simultaneous regulation of multiple genes. The parameters Ai, Am, and RA (3-oxohexanoyl-homoserine lactone [AHL] concentrations at which half of the maximum repression and the maximum repression were reached and value of the maximum repression when AHL was added manually, respectively) are defined and introduced to characterize repression curves, and the variant LuxRI58N is identified as the most suitable tuning factor for shake flask culture. Moreover, it is shown that dynamic overexpression of the Hfq chaperone is the key to combinatorial repression without disruptions on cell growth. To show a broad applicability, the production titers of pinene, pentalenene, and psilocybin are improved by 365.3%, 79.5%, and 302.9%, respectively, by applying combinatorial dynamic repression.


Asunto(s)
Escherichia coli/genética , Sitios Genéticos , Percepción de Quorum/genética , ARN Bacteriano/metabolismo , Monoterpenos Bicíclicos/metabolismo , Vías Biosintéticas/genética , Ciclopentanos/metabolismo , Regulación Bacteriana de la Expresión Génica , Glucólisis , Psilocibina/metabolismo
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