Expression of the Butyrivibrio fibrisolvens endo-β-1,4-glucanase gene together with the Erwinia pectate lyase and polygalacturonase genes in Saccharomyces cerevisiae

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dc.contributor.authorVan Rensburg P.
dc.contributor.authorVan Zyl W.H.
dc.contributor.authorPretorius I.S.
dc.date.accessioned2011-05-15T16:01:30Z
dc.date.available2011-05-15T16:01:30Z
dc.date.issued1994
dc.description.abstractRecombinant Saccharomyces cerevisiae strains capable of simultaneous secretion of bacterial glucanase and pectinase enzymes have been developed. The Butyrivibrio fibrisolvens endo-β-1,4-glucanase gene (end1), the Erwinia chrysanthemi pectate lyase gene (pe1E) and E. carotovora polygalacturonase gene (peh1) were each inserted between a yeast expression-secretion cassette and yeast gene terminator, and cloned into yeast-centromeric shuttle vectors. Transcription initiation signals present in the expression-secretion cassette were derived from the yeast alcohol dehydrogenase gene promoter (ADC1(p)), whereas the transcription termination signals were derived from the yeast tryptophan synthase gene terminator (TRP5(T)). Secretion of glucanase and pectinases was directed by the signal sequence of the yeast mating pheromone α-factor (MFα1(s)) These YCplac111-based constructs, designated END1, PEL5, and PEH1, respectively, were transformed into S. cerevisiae. The END1, PEL5 and PEH1 constructs were co-expressed in laboratory strains of S. cerevisiae as well as in wine and distillers' yeasts. DNA-RNA hybridization analysis showed the presence of END1, PEL5 and PEH1 transcripts. Carboxymethylcellulose and polypectate agarose assays revealed the production of biologically active endo-β-1,4-glucanase, pectate lyase and polygalacturonase by the S. cerevisiae transformants. Interestingly, although the same expression-secretion cassette was used in all three constructs, time-course assays indicated that the pectinases were secreted before the glucanase. It is tempting to speculate that the bulkiness of the END1-encoded protein and the five alternating repeats of Pro-Asp-Pro-Thr(Gln)-Pro-Val-Asp within the glucanase moiety could be involved in the delayed secretion of the glucanase.
dc.description.versionArticle
dc.identifier.citationCurrent Genetics
dc.identifier.citation27
dc.identifier.citation1
dc.identifier.issn1728083
dc.identifier.other10.1007/BF00326573
dc.identifier.urihttp://hdl.handle.net/10019.1/12008
dc.subjectcellulase
dc.subjectglucan glucosidase
dc.subjectpectate lyase
dc.subjectpolygalacturonase
dc.subjectarticle
dc.subjectbiotechnology
dc.subjectbiotransformation
dc.subjectdna rna hybridization
dc.subjecterwinia
dc.subjectfermentation
dc.subjectgene expression
dc.subjectgenetic engineering
dc.subjectnonhuman
dc.subjectpriority journal
dc.subjectpromoter region
dc.subjectrecombinant plasmid
dc.subjectsaccharomyces cerevisiae
dc.subjectshuttle vector
dc.subjecttranscription initiation
dc.subjecttranscription termination
dc.subjectyeast
dc.subjectAmino Acid Sequence
dc.subjectBacterial Proteins
dc.subjectBacteroidaceae
dc.subjectBase Sequence
dc.subjectCellulase
dc.subjectCellulose
dc.subjectErwinia
dc.subjectGene Expression
dc.subjectGenetic Vectors
dc.subjectIndustrial Microbiology
dc.subjectMolecular Sequence Data
dc.subjectPectins
dc.subjectPolygalacturonase
dc.subjectPolysaccharide-Lyases
dc.subjectRecombinant Fusion Proteins
dc.subjectSaccharomyces cerevisiae
dc.subjectSupport, Non-U.S. Gov't
dc.subjectBacteria (microorganisms)
dc.subjectButyrivibrio fibrisolvens
dc.subjectErwinia
dc.subjectErwinia chrysanthemi
dc.subjectSaccharomyces cerevisiae
dc.titleExpression of the Butyrivibrio fibrisolvens endo-β-1,4-glucanase gene together with the Erwinia pectate lyase and polygalacturonase genes in Saccharomyces cerevisiae
dc.typeArticle
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