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dc.contributor.authorWestereng, Bjørge
dc.contributor.authorLoose, Jennifer Sarah Maria
dc.contributor.authorVaaje-Kolstad, Gustav
dc.contributor.authorAachmann, Finn Lillelund
dc.contributor.authorSørlie, Morten
dc.contributor.authorEijsink, Vincent
dc.date.accessioned2019-04-02T12:39:29Z
dc.date.available2019-04-02T12:39:29Z
dc.date.created2018-09-24T07:28:58Z
dc.date.issued2018
dc.identifier.citationMethods in molecular biology. 2018, 1796 219-246.nb_NO
dc.identifier.issn1064-3745
dc.identifier.urihttp://hdl.handle.net/11250/2592960
dc.description.abstractLytic polysaccharide monooxygenases are copper-dependent enzymes that perform oxidative cleavage of glycosidic bonds in cellulose and various other polysaccharides. LPMOs acting on cellulose use a reactive oxygen species to abstract a hydrogen from the C1 or C4, followed by hydroxylation of the resulting substrate radical. The resulting hydroxylated species is unstable, resulting in glycoside bond scission and formation of an oxidized new chain end. These oxidized chain ends are spontaneously hydrated at neutral pH, leading to formation of an aldonic acid or a gemdiol, respectively. LPMO activity may be characterized using a variety of analytic tools, the most common of which are high-performance anion exchange chromatography system with pulsed amperometric detection (HPAEC-PAD) and MALDI-TOF mass spectrometry (MALDI-MS). NMR may be used to increase the certainty of product identifications, in particular the site of oxidation. Kinetic studies of LPMOs have several pitfalls and to avoid these, it is important to secure copper saturation, avoid the presence of free transition metals in solution, and control the amount of reductant (i.e., electron supply to the LPMO). Further insight into LPMO properties may be obtained by determining the redox potential and by determining the affinity for copper. In some cases, substrate affinity can be assessed using isothermal titration calorimetry. These methods are described in this chapter.nb_NO
dc.language.isoengnb_NO
dc.publisherHumana Pressnb_NO
dc.titleAnalytical Tools for Characterizing Cellulose-Active Lytic Polysaccharide Monooxygenases (LPMOs)nb_NO
dc.typeJournal articlenb_NO
dc.typePeer reviewednb_NO
dc.description.versionacceptedVersionnb_NO
dc.source.pagenumber219-246nb_NO
dc.source.volume1796nb_NO
dc.source.journalMethods in molecular biologynb_NO
dc.identifier.doi10.1007/978-1-4939-7877-9_16
dc.identifier.cristin1612711
dc.relation.projectNorges forskningsråd: 249865nb_NO
dc.relation.projectNorges forskningsråd: 226247nb_NO
dc.relation.projectNorges forskningsråd: 214613nb_NO
dc.relation.projectNorges forskningsråd: 244259nb_NO
dc.relation.projectNorges forskningsråd: 270038nb_NO
dc.description.localcodeThis is a post-peer-review, pre-copyedit version of an article published in Methods in molecular biology Locked until 1.6.2019 due to copyright restrictions. The final authenticated version is available online at: https://doi.org/10.1007/978-1-4939-7877-9_16nb_NO
cristin.unitcode194,66,15,0
cristin.unitnameInstitutt for bioteknologi og matvitenskap
cristin.ispublishedtrue
cristin.fulltextpostprint
cristin.qualitycode1


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