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dc.date.accessioned2020-04-06T18:23:48Z
dc.date.available2020-04-06T18:23:48Z
dc.date.created2019-09-24T07:49:36Z
dc.date.issued2020
dc.identifier.citationvan Eerde, André Varnai, Aniko Jameson, John-Kristian Paruch, Lisa Moen, Anders Anonsen, Jan Haug Chylenski, Piotr Steen, Hege Heldal, Inger Bock, Ralph Eijsink, Vincent Clarke, Jihong Liu . In-depth characterization of Trichoderma reesei cellobiohydrolase TrCel7A produced in Nicotiana benthamiana reveals limitations of cellulase production in plants by host-specific post-translational modifications. Plant Biotechnology Journal. 2019
dc.identifier.urihttp://hdl.handle.net/10852/74392
dc.description.abstractSustainable production of biofuels from lignocellulose feedstocks depends on cheap enzymes for degradation of such biomass. Plants offer a safe and cost‐effective production platform for biopharmaceuticals, vaccines and industrial enzymes boosting biomass conversion to biofuels. Production of intact and functional protein is a prerequisite for large‐scale protein production, and extensive host‐specific post‐translational modifications (PTMs) often affect the catalytic properties and stability of recombinant enzymes. Here we investigated the impact of plant PTMs on enzyme performance and stability of the major cellobiohydrolase TrCel7A from Trichoderma reesei, an industrially relevant enzyme. TrCel7A was produced in Nicotiana benthamiana using a vacuum‐based transient expression technology, and this recombinant enzyme (TrCel7Arec) was compared with the native fungal enzyme (TrCel7Anat) in terms of PTMs and catalytic activity on commercial and industrial substrates. We show that the N‐terminal glutamate of TrCel7Arec was correctly processed by N. benthamiana to a pyroglutamate, critical for protein structure, while the linker region of TrCel7Arec was vulnerable to proteolytic digestion during protein production due to the absence of O‐mannosylation in the plant host as compared with the native protein. In general, the purified full‐length TrCel7Arec had 25% lower catalytic activity than TrCel7Anat and impaired substrate‐binding properties, which can be attributed to larger N‐glycans and lack of O‐glycans in TrCel7Arec. All in all, our study reveals that the glycosylation machinery of N. benthamiana needs tailoring to optimize the production of efficient cellulases.
dc.languageEN
dc.rightsAttribution-NonCommercial-NoDerivatives 4.0 International
dc.rights.urihttps://creativecommons.org/licenses/by-nc-nd/4.0/
dc.titleIn-depth characterization of Trichoderma reesei cellobiohydrolase TrCel7A produced in Nicotiana benthamiana reveals limitations of cellulase production in plants by host-specific post-translational modifications
dc.typeJournal article
dc.creator.authorvan Eerde, André
dc.creator.authorVarnai, Aniko
dc.creator.authorJameson, John-Kristian
dc.creator.authorParuch, Lisa
dc.creator.authorMoen, Anders
dc.creator.authorAnonsen, Jan Haug
dc.creator.authorChylenski, Piotr
dc.creator.authorSteen, Hege
dc.creator.authorHeldal, Inger
dc.creator.authorBock, Ralph
dc.creator.authorEijsink, Vincent
dc.creator.authorClarke, Jihong Liu
cristin.unitcode185,15,29,40
cristin.unitnameSeksjon for biokjemi og molekylærbiologi
cristin.ispublishedtrue
cristin.fulltextoriginal
cristin.qualitycode1
dc.identifier.cristin1728096
dc.identifier.bibliographiccitationinfo:ofi/fmt:kev:mtx:ctx&ctx_ver=Z39.88-2004&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.jtitle=Plant Biotechnology Journal&rft.volume=&rft.spage=&rft.date=2019
dc.identifier.jtitlePlant Biotechnology Journal
dc.identifier.volume18
dc.identifier.issue3
dc.identifier.startpage631
dc.identifier.endpage643
dc.identifier.pagecount13
dc.identifier.doihttps://doi.org/10.1111/pbi.13227
dc.identifier.urnURN:NBN:no-77500
dc.type.documentTidsskriftartikkel
dc.type.peerreviewedPeer reviewed
dc.source.issn1467-7644
dc.identifier.fulltextFulltext https://www.duo.uio.no/bitstream/handle/10852/74392/2/2019_10.1111-pbi.13227.pdf
dc.type.versionPublishedVersion
dc.relation.projectNFR/257622
dc.relation.projectNFR/256766
dc.relation.projectNFR/243974


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