Heterologous expression of a Clostridium minicellulosome in Saccharomyces cerevisiae

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dc.contributor.authorLilly M.
dc.contributor.authorFierobe H.-P.
dc.contributor.authorVan Zyl W.H.
dc.contributor.authorVolschenk H.
dc.date.accessioned2011-05-15T16:03:26Z
dc.date.available2011-05-15T16:03:26Z
dc.date.issued2009
dc.description.abstractThe yeast Saccharomyces cerevisiae was genetically modified to assemble a minicellulosome on its cell surface by heterologous expression of a chimeric scaffoldin protein from Clostridium cellulolyticum under the regulation of the phosphoglycerate kinase 1 (PGK1) promoter and terminator regulatory elements, together with the β-xylanase 2 secretion signal of Trichoderma reesei and cell wall protein 2 (Cwp2) of S. cerevisiae. Fluorescent microscopy and Far Western blot analysis confirmed that the Scaf3p is targeted to the yeast cell surface and that the Clostridium thermocellum cohesin domain is functional in yeast. Similarly, functionality of the C. thermocellum dockerin domain in yeast is shown by binding to the Scaf3 protein in Far Western blot analysis. Phenotypic evidence for cohesin-dockerin interaction was also established with the detection of a twofold increase in tethered endoglucanase enzyme activity in S. cerevisiae cells expressing the Scaf3 protein compared with the parent strain. This study highlights the feasibility to future design of enhanced cellulolytic strains of S. cerevisiae through emulation of the cellulosome concept. Potentially, Scaf3p-armed yeast could also be developed into an alternative cell surface display strategy with various tailor-made applications. © 2009 Federation of European Microbiological Societies.
dc.description.versionArticle
dc.identifier.citationFEMS Yeast Research
dc.identifier.citation9
dc.identifier.citation8
dc.identifier.issn15671356
dc.identifier.other10.1111/j.1567-1364.2009.00564.x
dc.identifier.urihttp://hdl.handle.net/10019.1/12621
dc.subjectcell membrane protein
dc.subjectcell membrane protein 2
dc.subjectcellulosome
dc.subjectcohesin
dc.subjectendo 1,4 beta xylanase
dc.subjectfungal protein
dc.subjectglucan synthase
dc.subjectphosphoglycerate kinase
dc.subjectphosphoglycerate kinase 1
dc.subjectprotein dockerin
dc.subjectprotein scaf3
dc.subjectunclassified drug
dc.subjectarticle
dc.subjectClostridium
dc.subjectClostridium cellulolyticum
dc.subjectClostridium thermocellum
dc.subjectcontrolled study
dc.subjectenzyme activity
dc.subjectenzyme regulation
dc.subjectheterologous expression
dc.subjectnonhuman
dc.subjectprotein assembly
dc.subjectprotein binding
dc.subjectprotein domain
dc.subjectprotein function
dc.subjectprotein interaction
dc.subjectprotein targeting
dc.subjectSaccharomyces cerevisiae
dc.subjectsignal transduction
dc.subjectBacterial Proteins
dc.subjectBlotting, Far-Western
dc.subjectCellulase
dc.subjectCellulose
dc.subjectCellulosomes
dc.subjectClostridium thermocellum
dc.subjectMembrane Proteins
dc.subjectMicroscopy, Fluorescence
dc.subjectPromoter Regions, Genetic
dc.subjectProtein Binding
dc.subjectProtein Sorting Signals
dc.subjectRecombinant Proteins
dc.subjectSaccharomyces cerevisiae
dc.subjectTrichoderma
dc.subjectClostridium
dc.subjectClostridium cellulolyticum
dc.subjectClostridium thermocellum
dc.subjectHypocrea jecorina
dc.subjectSaccharomyces cerevisiae
dc.titleHeterologous expression of a Clostridium minicellulosome in Saccharomyces cerevisiae
dc.typeArticle
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