Influence of a family 29 carbohydrate binding module on the activity of galactose oxidase from Fusarium graminearum

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dc.contributorAalto-yliopistofi
dc.contributorAalto Universityen
dc.contributor.authorMollerup, Filip
dc.contributor.authorParikka, Kirsti
dc.contributor.authorVuong, Thu V.
dc.contributor.authorTenkanen, Maija
dc.contributor.authorMaster, Emma
dc.contributor.departmentDepartment of Biotechnology and Chemical Technology
dc.contributor.departmentUniversity of Helsinki
dc.contributor.departmentUniversity of Toronto
dc.contributor.departmentDepartment of Bioproducts and Biosystemsen
dc.date.accessioned2019-01-14T09:19:12Z
dc.date.available2019-01-14T09:19:12Z
dc.date.embargoinfo:eu-repo/date/embargoEnd/2018-02-02
dc.date.issued2016-02-01
dc.description| openaire: EC/H2020/33947/EU//BHIVE
dc.description.abstractBackground Galactose oxidase (GaO) selectively oxidizes the primary hydroxyl of galactose to a carbonyl, facilitating targeted chemical derivatization of galactose-containing polysaccharides, leading to renewable polymers with tailored physical and chemical properties. Here we investigate the impact of a family 29 glucomannan binding module on the activity and binding of GaO towards various polysaccharides. Specifically, CBM29-1-2 from Piromyces equi was separately linked to the N- and C-termini of GaO. Results Both GaO-CBM29 and CBM29-GaO were successfully expressed in Pichia pastoris, and demonstrated enhanced binding to galactomannan, galactoglucomannan and galactoxyloglucan. The position of the CBM29 fusion affected the enzyme function. Particularly, C-terminal fusion led to greatest increases in galactomannan binding and catalytic efficiency, where relative to wild-type GaO, kcat/Km values increased by 7.5 and 19.8 times on guar galactomannan and locust bean galactomannan, respectively. The fusion of CBM29 also induced oligomerization of GaO-CBM29. Major conclusions Similar to impacts of cellulose-binding modules associated with cellulolytic enzymes, increased substrate binding impeded the action of GaO fusions on more concentrated preparations of galactomannan, galactoglucomannan and galactoxyloglucan; this was especially true for GaO-CBM29. Given the N-terminal positioning of the native galactose-binding CBM32 in GaO, the varying impacts of N-terminal versus C-terminal fusion of CBM29-1-2 may reflect competing action of neighboring CBMs. General significance This study thoroughly examines and discusses the effects of CBM fusion to non-lignocellulytic enzymes on soluble polysaccharides. Herein kinetics of GaO on galactose containing polysaccharides is presented for the first time.en
dc.description.versionPeer revieweden
dc.format.extent9
dc.format.extent354-362
dc.format.mimetypeapplication/pdf
dc.identifier.citationMollerup , F , Parikka , K , Vuong , T V , Tenkanen , M & Master , E 2016 , ' Influence of a family 29 carbohydrate binding module on the activity of galactose oxidase from Fusarium graminearum ' , Biochimica et Biophysica Acta: General Subjects , vol. 1860 , no. 2 , pp. 354-362 . https://doi.org/10.1016/j.bbagen.2015.10.023en
dc.identifier.doi10.1016/j.bbagen.2015.10.023
dc.identifier.issn0304-4165
dc.identifier.otherPURE UUID: 18a93add-dc2a-4018-832c-ca4579a4511a
dc.identifier.otherPURE ITEMURL: https://research.aalto.fi/en/publications/18a93add-dc2a-4018-832c-ca4579a4511a
dc.identifier.otherPURE LINK: http://www.scopus.com/inward/record.url?scp=84948469936&partnerID=8YFLogxK
dc.identifier.otherPURE FILEURL: https://research.aalto.fi/files/31095561/CHEM_ERC_Mollerup_Master_et_al_Influence_of_a_family_29_2016_BIOCHIMICA.pdf
dc.identifier.urihttps://aaltodoc.aalto.fi/handle/123456789/35914
dc.identifier.urnURN:NBN:fi:aalto-201901141097
dc.language.isoenen
dc.relationinfo:eu-repo/grantAgreement/EC/H2020/33947/EU//BHIVE
dc.relation.ispartofseriesBIOCHIMICA ET BIOPHYSICA ACTA: GENERAL SUBJECTSen
dc.relation.ispartofseriesVolume 1860, issue 2en
dc.rightsopenAccessen
dc.subject.keywordCarbohydrate binding module
dc.subject.keywordFusion proteins
dc.subject.keywordGalactoglucomannan
dc.subject.keywordGalactomannan
dc.subject.keywordGalactose oxidase
dc.subject.keywordGalactoxyloglucan
dc.titleInfluence of a family 29 carbohydrate binding module on the activity of galactose oxidase from Fusarium graminearumen
dc.typeA1 Alkuperäisartikkeli tieteellisessä aikakauslehdessäfi
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