Production of soluble regulatory hydrogenase from Ralstonia eutropha in Escherichia coli using a fed-batch-based autoinduction system

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dc.contributor.authorFan, Qi
dc.contributor.authorNeubauer, Peter
dc.contributor.authorGimpel, Matthias
dc.date.accessioned2021-12-13T07:42:13Z
dc.date.available2021-12-13T07:42:13Z
dc.date.issued2021-10-18
dc.description.abstractBackground: Autoinduction systems can regulate protein production in Escherichia coli without the need to monitor cell growth or add inducer at the proper time following culture growth. Compared to classical IPTG induction, autoinduction provides a simple and fast way to obtain high protein yields. In the present study, we report on the optimization process for the enhanced heterologous production of the Ralstonia eutropha regulatory hydrogenase (RH) in E. coli using autoinduction. These autoinduction methods were combined with the EnPresso B fed-batch like growth system, which applies slow in situ enzymatic glucose release from a polymer to control cell growth and protein synthesis rate. Results: We were able to produce 125 mg L−1 RH corresponding to a productivity averaged over the whole process time of 3 mg (L h)−1 in shake flasks using classic single-shot IPTG induction. IPTG autoinduction resulted in a comparable volumetric RH yield of 112 mg L−1 and due to the shorter overall process time in a 1.6-fold higher productivity of 5 mg (L h)−1. In contrast, lactose autoinduction increased the volumetric yield more than 2.5-fold and the space time yield fourfold reaching 280 mg L−1 and 11.5 mg (L h)−1, respectively. Furthermore, repeated addition of booster increased RH production to 370 mg L−1, which to our knowledge is the highest RH concentration produced in E. coli to date. Conclusions: The findings of this study confirm the general feasibility of the developed fed-batch based autoinduction system and provide an alternative to conventional induction systems for efficient recombinant protein production. We believe that the fed-batch based autoinduction system developed herein will favor the heterologous production of larger quantities of difficult-to-express complex enzymes to enable economical production of these kinds of proteins.en
dc.description.sponsorshipDFG, 390540038, EXC 2008: Unifying Systems in Catalysis "UniSysCat"en
dc.description.sponsorshipTU Berlin, Open-Access-Mittel – 2021en
dc.identifier.eissn1475-2859
dc.identifier.pmid34663324
dc.identifier.urihttps://depositonce.tu-berlin.de/handle/11303/14037
dc.identifier.urihttp://dx.doi.org/10.14279/depositonce-12810
dc.language.isoenen
dc.relation.ispartof10.14279/depositonce-16438
dc.rights.urihttps://creativecommons.org/licenses/by/4.0/en
dc.subject.ddc540 Chemie und zugeordnete Wissenschaftende
dc.subject.otherEscherichia colien
dc.subject.otherralstonia eutrophaen
dc.subject.otherdifficult-to-express proteinen
dc.subject.otherEnPresso Ben
dc.subject.otherfed-batch like cultivationen
dc.subject.otherIPTG autoinductionen
dc.subject.otherlactose autoinductionen
dc.subject.otherregulatory hydrogenaseen
dc.titleProduction of soluble regulatory hydrogenase from Ralstonia eutropha in Escherichia coli using a fed-batch-based autoinduction systemen
dc.typeArticleen
dc.type.versionpublishedVersionen
dcterms.bibliographicCitation.articlenumber201en
dcterms.bibliographicCitation.doi10.1186/s12934-021-01690-4en
dcterms.bibliographicCitation.journaltitleMicrobial Cell Factoriesen
dcterms.bibliographicCitation.originalpublishernameSpringer Natureen
dcterms.bibliographicCitation.originalpublisherplaceLondonen
dcterms.bibliographicCitation.volume20en
tub.accessrights.dnbfreeen
tub.affiliationFak. 3 Prozesswissenschaften::Inst. Biotechnologie::FG Bioverfahrenstechnikde
tub.affiliation.facultyFak. 3 Prozesswissenschaften
tub.affiliation.groupFG Bioverfahrenstechnik
tub.affiliation.instituteInst. Biotechnologie
tub.publisher.universityorinstitutionTechnische Universität Berlinen

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