Transport-controlled growth decoupling for self-induced protein expression with a glycerol-repressible genetic circuit

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dc.authoridLara, Alvaro/0000-0003-3535-7619|Delvigne, Frank/0000-0002-1679-1914
dc.authorwosidLara, Alvaro/A-8397-2012
dc.contributor.authorLara, Alvaro R.
dc.contributor.authorKunert, Flavio
dc.contributor.authorVandenbroucke, Vincent
dc.contributor.authorTaymaz-Nikerel, Hilal
dc.contributor.authorMartinez, Luz Maria
dc.contributor.authorSigala, Juan-Carlos
dc.contributor.authorDelvigne, Frank
dc.date.accessioned2024-07-18T20:40:01Z
dc.date.available2024-07-18T20:40:01Z
dc.date.issued2024
dc.departmentİstanbul Bilgi Üniversitesien_US
dc.description.abstractDecoupling cell formation from recombinant protein synthesis is a potent strategy to intensify bioprocesses. Escherichia coli strains with mutations in the glucose uptake components lack catabolite repression, display low growth rate, no overflow metabolism, and high recombinant protein yields. Fast growth rates were promoted by the simultaneous consumption of glucose and glycerol, and this was followed by a phase of slow growth, when only glucose remained in the medium. A glycerol-repressible genetic circuit was designed to autonomously induce recombinant protein expression. The engineered strain bearing the genetic circuit was cultured in 3.9 g L-1 glycerol + 18 g L-1 glucose in microbioreactors with online oxygen transfer rate monitoring. The growth was fast during the simultaneous consumption of both carbon sources (C-sources), while expression of the recombinant protein was low. When glycerol was depleted, the growth rate decreased, and the specific fluorescence reached values 17% higher than those obtained with a strong constitutive promoter. Despite the relatively high amount of C-source used, no oxygen limitation was observed. The proposed approach eliminates the need for the substrate feeding or inducers addition and is set as a simple batch culture while mimicking fed-batch performance. The authors propose a strategy that combines cell engineering and genetic circuit design to decouple growth rate from recombinant protein expression based on the carbon source transport. A simple batch scheme allows autoinduction and growth rate shift, mimicking a fed-batch mode. The results are potentially useful for product and process development. imageen_US
dc.description.sponsorshipConsejo Nacional de Humanidades, Ciencias y Tecnologas [A1-S-8646]; Exploratory Research Space, RWTH Aachen Universityen_US
dc.description.sponsorshipThis work was supported by CONAHCyT grant A1-S-8646 and the Exploratory Research Space, RWTH Aachen University.en_US
dc.identifier.doi10.1002/bit.28697
dc.identifier.issn0006-3592
dc.identifier.issn1097-0290
dc.identifier.pmid38470342en_US
dc.identifier.scopus2-s2.0-85187488856en_US
dc.identifier.scopusqualityQ1en_US
dc.identifier.urihttps://doi.org/10.1002/bit.28697
dc.identifier.urihttps://hdl.handle.net/11411/6930
dc.identifier.wosWOS:001182209300001en_US
dc.identifier.wosqualityN/Aen_US
dc.indekslendigikaynakWeb of Scienceen_US
dc.indekslendigikaynakScopusen_US
dc.indekslendigikaynakPubMeden_US
dc.language.isoenen_US
dc.publisherWileyen_US
dc.relation.ispartofBiotechnology and Bioengineeringen_US
dc.relation.publicationcategoryMakale - Uluslararası Hakemli Dergi - Kurum Öğretim Elemanıen_US
dc.rightsinfo:eu-repo/semantics/openAccessen_US
dc.subjectBioprocess İntensificationen_US
dc.subjectC-Source Mixturesen_US
dc.subjectGenetic Circuiten_US
dc.subjectGrowth Decouplingen_US
dc.subjectMicrobial Engineeringen_US
dc.subjectToggle Switchen_US
dc.subjectGlucoseen_US
dc.subjectMetabolismen_US
dc.subjectSystemen_US
dc.titleTransport-controlled growth decoupling for self-induced protein expression with a glycerol-repressible genetic circuiten_US
dc.typeArticleen_US

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