Heterologous co-production of Thermobifida fusca Cel9A with other cellulases in Saccharomyces cerevisiae

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dc.contributor.authorVan Wyk N.
dc.contributor.authorDen Haan R.
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 processive endoglucanase Cel9A of the moderately thermophilic actinomycete Thermobifida fusca was functionally produced in Saccharomyces cerevisiae. Recombinant Cel9A displayed activity on both soluble (carboxymethylcellulose) and insoluble (Avicel) cellulose substrates confirming its processive endoglucanase activity. High-performance anionic exchange chromatography analyses of soluble sugars released from Avicel revealed a cellobiose/glucose ratio of 2.5∈±∈0.1. Growth by the recombinant strain on amorphous cellulose was possible due to the sufficient amount of glucose cleaved from the cellulose chain. This is the first confirmed report of S. cerevisiae growing on a cellulosic substrate as sole carbohydrate source while only expressing one recombinant gene. To improve the cellulolytic capability of S. cerevisiae and to investigate the level of synergy among cellulases produced by a recombinant host, the cel9A gene was co-expressed with four cellulase-coding genes of Trichoderma reesei: two endoglucanases cel5A (egII) and cel7B (egI), and two cellobiohydrolases cel6A (cbhII) and cel7A (cbhI). Synergy, especially between the Cel9A and the two cellobiohydrolases, resulted in a higher cellulolytic capability of the recombinant host. © 2010 Springer-Verlag.
dc.description.versionArticle
dc.identifier.citationApplied Microbiology and Biotechnology
dc.identifier.citation87
dc.identifier.citation5
dc.identifier.issn1757598
dc.identifier.other10.1007/s00253-010-2618-z
dc.identifier.urihttp://hdl.handle.net/10019.1/9931
dc.subjectAmorphous cellulose
dc.subjectAnionic exchange
dc.subjectCarbohydrate sources
dc.subjectCarboxy methylcellulose
dc.subjectCellobiohydrolases
dc.subjectCellulose chain
dc.subjectCellulose substrates
dc.subjectCellulosic substrates
dc.subjectCo-expression
dc.subjectCo-production
dc.subjectCoding genes
dc.subjectConsolidated bioprocessing
dc.subjectEndoglucanases
dc.subjectModerately thermophilic
dc.subjectPhosphoric-acid swollen cellulose
dc.subjectRecombinant hosts
dc.subjectRecombinant strains
dc.subjectS.cerevisiae
dc.subjectSaccharomyces cerevisiae
dc.subjectSoluble sugars
dc.subjectThermobifida fusca
dc.subjectTrichoderma reesei
dc.subjectAcids
dc.subjectCarbohydrates
dc.subjectCellulose
dc.subjectGlucose
dc.subjectPhosphoric acid
dc.subjectSubstrates
dc.subjectSugar (sucrose)
dc.subjectYeast
dc.subjectcarboxymethylcellulose
dc.subjectcellobiose
dc.subjectcellulase
dc.subjectglucose
dc.subjectbacterium
dc.subjectcellulose
dc.subjectchromatography
dc.subjectenzyme activity
dc.subjectgene expression
dc.subjectgrowth rate
dc.subjectrecombination
dc.subjectsugar
dc.subjectyeast
dc.subjectarticle
dc.subjectbacterial gene
dc.subjectbacterial growth
dc.subjectbacterial strain
dc.subjectcel5a gene
dc.subjectcel6a gene
dc.subjectcel7a gene
dc.subjectCel9a gene
dc.subjectenzyme activity
dc.subjectgene expression
dc.subjectHypocrea jecorina
dc.subjectnonhuman
dc.subjectprotein cleavage
dc.subjectprotein glycosylation
dc.subjectSaccharomyces cerevisiae
dc.subjectsoil microflora
dc.subjectThermobifida fusca
dc.subjectActinomycetales
dc.subjectBacterial Proteins
dc.subjectCellulases
dc.subjectCellulose
dc.subjectChromatography, Ion Exchange
dc.subjectFungal Proteins
dc.subjectGlucose
dc.subjectRecombinant Proteins
dc.subjectSaccharomyces cerevisiae
dc.subjectTrichoderma
dc.subjectAcids
dc.subjectCarbohydrates
dc.subjectCellulase
dc.subjectCellulose
dc.subjectGlucose
dc.subjectPhosphoric Acid
dc.subjectSaccharomyces Cerevisiae
dc.subjectSubstrates
dc.subjectSugar
dc.subjectHypocrea jecorina
dc.subjectSaccharomyces cerevisiae
dc.subjectThermobifida fusca
dc.titleHeterologous co-production of Thermobifida fusca Cel9A with other cellulases in Saccharomyces cerevisiae
dc.typeArticle
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