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dc.contributor.authorvan Eerde, André
dc.contributor.authorVarnai, Aniko
dc.contributor.authorJameson, John-Kristian
dc.contributor.authorParuch, Lisa
dc.contributor.authorMoen, Anders
dc.contributor.authorAnonsen, Jan Haug
dc.contributor.authorChylenski, Piotr
dc.contributor.authorSteen, Hege
dc.contributor.authorHeldal, Inger
dc.contributor.authorBock, Ralph
dc.contributor.authorEijsink, Vincent
dc.contributor.authorClarke, Jihong Liu
dc.identifier.citationPlant Biotechnology Journal, 2019.nb_NO
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.nb_NO
dc.rightsAttribution-NonCommercial-NoDerivatives 4.0 Internasjonal*
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 modificationsnb_NO
dc.typeJournal articlenb_NO
dc.typePeer reviewednb_NO
dc.source.journalPlant Biotechnology Journalnb_NO
dc.relation.projectNorges forskningsråd: 257622nb_NO
dc.relation.projectNorges forskningsråd: 256766nb_NO
dc.relation.projectNorges forskningsråd: 243974nb_NO
cristin.unitnameKjemi, bioteknologi og matvitenskap

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Attribution-NonCommercial-NoDerivatives 4.0 Internasjonal
Med mindre annet er angitt, så er denne innførselen lisensiert som Attribution-NonCommercial-NoDerivatives 4.0 Internasjonal