High-level heterologous expression of active Chaetomium thermophilum FDH in Pichia pastoris

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dc.contributor.authorDuman, Zeynep Efsun
dc.contributor.authorDuraksoy, Bedri Burak
dc.contributor.authorAktaş, Fatih
dc.contributor.authorWoodley, John M.
dc.contributor.authorBinay, Barış
dc.date.accessioned2020-04-30T13:32:33Z
dc.date.available2020-04-30T13:32:33Z
dc.date.issued2020
dc.departmentDÜ, Mühendislik Fakültesi, Endüstri Mühendisliği Bölümüen_US
dc.description.abstractNowadays, the use of formate dehydrogenase (FDH, EC 1.17.1.9) is well established as a means of NADH regeneration from NAD+ via the coupled conversion of formate into carbon dioxide. Recent studies have been reported that specifically Chaetomium thermophilum FDH (CtFDH) is the most efficient FDH catalyzing this reaction in reverse (i.e. using CO2 as a substrate to produce formate, and thereby regenerating NAD+). However, to date the production of active CtFDH at high protein expression levels has received relatively little attention. In this study, we have tested the effect of batch and high cell density fermentation (HCDF) strategies in a small stirred fermenter, as well as the effect of supplementing the medium with casamino acids, on the expressed level of secreted CtFDH using P. pastoris. We have established that the amount of expressed CtFDH was indeed enhanced via a HCDF strategy and that extracellular protease activity was eliminated via the addition of casamino acids into the fermentation medium. On this basis, secreted CtFDH in an active form can be easily separated from the fermentation and can be used for subsequent biotechnological applications. © 2020 Elsevier Inc.en_US
dc.description.sponsorshipDüzce Üniversitesien_US
dc.description.sponsorshipThis work was supported by Düzce University Scientific Research Project Department with 2017.06.02.578 project number and special thanks to BERC Lab for providing p PICZ? A vector with 6xHis-tagged Ct FDH. The authors also gratefully acknowledge the help and support of Rowan M Lindeque (Department of Chemical and Biochemical Engineering, DTU, Denmark).en_US
dc.identifier.doi10.1016/j.enzmictec.2020.109552en_US
dc.identifier.issn0141-0229
dc.identifier.scopusqualityN/Aen_US
dc.identifier.urihttps://dx.doi.org/10.1016/j.enzmictec.2020.109552
dc.identifier.urihttps://hdl.handle.net/20.500.12684/346
dc.identifier.volume137en_US
dc.indekslendigikaynakScopusen_US
dc.language.isoenen_US
dc.publisherElsevier Inc.en_US
dc.relation.ispartofEnzyme and Microbial Technologyen_US
dc.relation.publicationcategoryMakale - Uluslararası Hakemli Dergi - Kurum Öğretim Elemanıen_US
dc.rightsinfo:eu-repo/semantics/closedAccessen_US
dc.subjectCasamino acid addition; CtFDH; HCDF strategy; P. pastoris expression systemen_US
dc.titleHigh-level heterologous expression of active Chaetomium thermophilum FDH in Pichia pastorisen_US
dc.typeArticleen_US

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