Exploring improved endoglucanase expression in Saccharomyces cerevisiae strains

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dc.contributor.authorDu Plessis L.
dc.contributor.authorRose S.H.
dc.contributor.authorVan Zyl W.H.
dc.date.accessioned2011-05-15T15:56:35Z
dc.date.available2011-05-15T15:56:35Z
dc.date.issued2010
dc.description.abstractThe endoglucanase I and II genes (egI or Cel7B and egII or Cel5A) of Trichoderma reesei QM6a were successfully cloned and expressed in Saccharomyces cerevisiae under the transcriptional control of the yeast ENO1 promoter and terminator sequences. Random mutagenesis of the egI-bearing plasmid resulted in a twofold increase in extracellular EGI activity. Both endoglucanase genes were co-expressed with the synthetic, codon-optimised cellobiohydrolase gene (s-cbhI) from T. reesei as well as the β-glucosidase gene (bgl1) from Saccharomycopsis fibuligera in S. cerevisiae. Extracellular endoglucanase activity was lower when co-expressed with s-cbhI or bgl1. Recombinant strains were able to hydrolyse phosphoric acid swollen cellulose, generating mainly cellotriose, cellobiose and glucose. Cellobiose accumulated, suggesting the β-glucosidase activity to be the rate-limiting factor. As a consequence, the recombinant strains were unable to produce enough glucose for growth on amorphous cellulose. The results of this study provide insight into further optimisation of recombinantly expressed cellulase combinations for saccharification and fermentation of cellulose to ethanol. © 2009 Springer-Verlag.
dc.description.versionArticle
dc.identifier.citationApplied Microbiology and Biotechnology
dc.identifier.citation86
dc.identifier.citation5
dc.identifier.issn1757598
dc.identifier.other10.1007/s00253-009-2403-z
dc.identifier.urihttp://hdl.handle.net/10019.1/9933
dc.subjectAmorphous cellulose
dc.subjectCellobiohydrolases
dc.subjectCellobiose
dc.subjectCellulose degradation
dc.subjectEndoglucanase gene
dc.subjectEndoglucanase I
dc.subjectEndoglucanases
dc.subjectExtracellular
dc.subjectGlucosidase
dc.subjectGlucosidase activity
dc.subjectOptimisations
dc.subjectPhosphoric-acid swollen cellulose
dc.subjectRandom mutagenesis
dc.subjectRate limiting factors
dc.subjectRecombinant strains
dc.subjectS.cerevisiae
dc.subjectSaccharomyces cerevisiae
dc.subjectSaccharomyces cerevisiae strains
dc.subjectSaccharomycopsis
dc.subjectT. reesei
dc.subjectTerminator sequences
dc.subjectTranscriptional control
dc.subjectTrichoderma reesei
dc.subjectBioethanol
dc.subjectDegradation
dc.subjectEthanol
dc.subjectGenes
dc.subjectGlucose
dc.subjectPhosphoric acid
dc.subjectSaccharification
dc.subjectYeast
dc.subjectCellulose
dc.subjectalcohol
dc.subjectbeta glucosidase
dc.subjectcellulose
dc.subjectcellulose 1,4 beta cellobiosidase
dc.subjectglucan synthase
dc.subjectglucose
dc.subjectphosphoric acid
dc.subjectcellulose
dc.subjectenzyme activity
dc.subjectethanol
dc.subjectfermentation
dc.subjectgene expression
dc.subjecthydrolysis
dc.subjectmutation
dc.subjectplasmid
dc.subjectrecombination
dc.subjectyeast
dc.subjectarticle
dc.subjectcontrolled study
dc.subjectenzyme activity
dc.subjectEscherichia coli
dc.subjectfermentation
dc.subjectfungal strain
dc.subjecthigh performance liquid chromatography
dc.subjecthydrolysis
dc.subjectHypocrea jecorina
dc.subjectmutagenesis
dc.subjectnonhuman
dc.subjectnucleotide sequence
dc.subjectplasmid
dc.subjectprotein expression
dc.subjectsaccharification
dc.subjectSaccharomyces cerevisiae
dc.subjectSaccharomycetales
dc.subjectAmino Acid Sequence
dc.subjectCellulase
dc.subjectCellulases
dc.subjectCellulose
dc.subjectChromatography, High Pressure Liquid
dc.subjectCloning, Molecular
dc.subjectEscherichia coli
dc.subjectEthanol
dc.subjectGene Dosage
dc.subjectGlucose
dc.subjectMolecular Sequence Data
dc.subjectMutagenesis
dc.subjectPlasmids
dc.subjectSaccharomyces cerevisiae
dc.subjectSaccharomyces cerevisiae Proteins
dc.subjectSaccharomycopsis
dc.subjectTrichoderma
dc.subjectBiodegradation
dc.subjectCellulose
dc.subjectCellulose Degradation
dc.subjectEndo Enzymes
dc.subjectEthanol
dc.subjectGenes
dc.subjectGlucanase
dc.subjectGlucose
dc.subjectPhosphoric Acid
dc.subjectSaccharification
dc.subjectSaccharomyces Cerevisiae
dc.subjectStrains
dc.subjectHypocrea jecorina
dc.subjectSaccharomyces cerevisiae
dc.subjectSaccharomycopsis fibuligera
dc.titleExploring improved endoglucanase expression in Saccharomyces cerevisiae strains
dc.typeArticle
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