Thermostable adenosine 5′-monophosphate phosphorylase from Thermococcus kodakarensis forms catalytically active inclusion bodies

dc.contributor.authorKamel, Sarah
dc.contributor.authorWalczak, Miriam C.
dc.contributor.authorKaspar, Felix
dc.contributor.authorWestarp, Sarah
dc.contributor.authorNeubauer, Peter
dc.contributor.authorKurreck, Anke
dc.date.accessioned2021-12-09T19:45:17Z
dc.date.available2021-12-09T19:45:17Z
dc.date.issued2021-08-19
dc.description.abstractCatalytically active inclusion bodies (CatIBs) produced in Escherichia coli are an interesting but currently underexplored strategy for enzyme immobilization. They can be purified easily and used directly as stable and reusable heterogenous catalysts. However, very few examples of CatIBs that are naturally formed during heterologous expression have been reported so far. Previous studies have revealed that the adenosine 5′-monophosphate phosphorylase of Thermococcus kodakarensis ( Tk AMPpase) forms large soluble multimers with high thermal stability. Herein, we show that heat treatment of soluble protein from crude extract induces aggregation of active protein which phosphorolyse all natural 5′-mononucleotides. Additionally, inclusion bodies formed during the expression in E. coli were found to be similarly active with 2–6 folds higher specific activity compared to these heat-induced aggregates. Interestingly, differences in the substrate preference were observed. These results show that the recombinant thermostable Tk AMPpase is one of rare examples of naturally formed CatIBs.en
dc.description.sponsorshipTU Berlin, Open-Access-Mittel – 2021en
dc.identifier.eissn2045-2322
dc.identifier.pmid34413335
dc.identifier.urihttps://depositonce.tu-berlin.de/handle/11303/14017
dc.identifier.urihttp://dx.doi.org/10.14279/depositonce-12790
dc.language.isoenen
dc.rights.urihttps://creativecommons.org/licenses/by/4.0/en
dc.subject.ddc500 Naturwissenschaften und Mathematikde
dc.subject.otherbiochemistryen
dc.subject.otherbiotechnologyen
dc.subject.othermicrobiologyen
dc.subject.otherE. colien
dc.subject.otherTkAMPpaseen
dc.subject.otherCatIBsen
dc.subject.otherenzymeen
dc.titleThermostable adenosine 5′-monophosphate phosphorylase from Thermococcus kodakarensis forms catalytically active inclusion bodiesen
dc.typeArticleen
dc.type.versionpublishedVersionen
dcterms.bibliographicCitation.articlenumber16880en
dcterms.bibliographicCitation.doi10.1038/s41598-021-96073-5en
dcterms.bibliographicCitation.issue1en
dcterms.bibliographicCitation.journaltitleScientific Reportsen
dcterms.bibliographicCitation.originalpublishernameSpringer Natureen
dcterms.bibliographicCitation.originalpublisherplaceLondonen
dcterms.bibliographicCitation.volume11en
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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