Expression of the major bean proteins from Theobroma cacao (cocoa) in the yeasts Hansenula polymorpha and Saccharomyces cerevisiae

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dc.authorscopusid55845419400
dc.authorscopusid9041842400
dc.authorscopusid6701537863
dc.authorscopusid57213482691
dc.authorscopusid7003616838
dc.contributor.authorYavuz M.Ö.
dc.contributor.authorAshton S.M.V.
dc.contributor.authorDeakin E.D.
dc.contributor.authorSpencer M.E.
dc.contributor.authorSudbery P.E.
dc.date.accessioned2024-06-12T10:24:58Z
dc.date.available2024-06-12T10:24:58Z
dc.date.issued1996
dc.description.abstractThe production in two yeast expression systems of recombinant forms of the major proteins from the cocoa bean is described. Three major protein species are found in the cocoa bean: an albumin of molecular mass 21 kDa (p21) and two insoluble vicilin-like proteins of molecular mass 31 kDa and 47 kDa (p31 and p47, respectively). The p31 and p47 species are known to be derived from a common 67-kDa precursor (p67) by post-translational processing that includes the deletion of a hydrophilic domain located immediately after an N-terminal signal sequence. All three proteins appear to be targeted to membrane-bound storage organelles by N-terminal signal sequences. The p21 and p67 coding sequences were expressed in Hansenula polymorpha using the powerful methanol oxidase (MOX) promoter and in Saccharomyces cerevisiae using the promoter of the pyruvate kinase (PYK) gene. The expression constructs contained the native plant signal sequence, or various yeast signals. The p21 protein was successfully expressed and secreted from both yeasts. The insoluble p67 protein proved more difficult. Species of the correct molecular mass were recovered internally and small amounts of a p47 species were secreted using a yeast leader sequence. However, proteolytic cleavage, probably due to Kex2p-like processing, led to the appearance of other protein species.The production in two yeast expression systems of recombinant forms of the major proteins from the cocoa bean is described. Three major protein species are found in the cocoa bean: an albumin of molecular mass 21 kDa (p21) and two insoluble vicilin-like proteins of molecular mass 31 kDa and 47 kDa (p31 and p47, respectively). The p31 and p47 species me known to be derived from a common 67-kDa precursor (p67) by post-translational processing that includes the deletion of a hydrophilic domain located immediately after an N-terminal signal sequence. All three proteins appear to be targeted to membrane-bound storage organelles by N-terminal signal sequences. The p21 and p67 coding sequences were expressed in Hansenula polymorpha using the powerful methanol oxidase (MOX) promoter and in Saccharomyces cerevisiae using the promoter of the pyruvate kinase (PYK) gene. The expression constructs contained the native plant signal sequence, or various yeast signals. The p21 protein was successfully expressed and secreted from both yeasts. The insoluble p67 protein proved more difficult. Species of the correct molecular mass were recovered internally and small amounts of a p47 species were secreted using a yeast leader sequence. However, proteolytic cleavage, probably due to Kex2p-like processing, led to the appearance of other protein species.en_US
dc.description.sponsorshipM.E. Spencer, E.A. Deakin and S.M.V. Ashton are very grateful for financial support from Mars Confectionery (Slough, UK) covering part of this project.en_US
dc.identifier.doi10.1016/0168-1656(95)00181-6
dc.identifier.endpage54en_US
dc.identifier.issn0168-1656
dc.identifier.issue1en_US
dc.identifier.pmid8672284en_US
dc.identifier.scopus2-s2.0-0343113395en_US
dc.identifier.scopusqualityQ2en_US
dc.identifier.startpage43en_US
dc.identifier.urihttps://doi.org/10.1016/0168-1656(95)00181-6
dc.identifier.urihttps://hdl.handle.net/20.500.14551/16128
dc.identifier.volume46en_US
dc.indekslendigikaynakScopusen_US
dc.indekslendigikaynakPubMeden_US
dc.language.isoenen_US
dc.publisherElsevier Science B.V., Amsterdamen_US
dc.relation.ispartofJournal of Biotechnologyen_US
dc.relation.publicationcategoryMakale - Uluslararası Hakemli Dergi - Kurum Öğretim Elemanıen_US
dc.rightsinfo:eu-repo/semantics/closedAccessen_US
dc.subjectHansenula Polymorpha; Mox Promoter; Pyk Promoter; Recombinant Cocoa Protein; Saccharomyces Cerevisiae; Theobroma Cacaoen_US
dc.subjectBiotechnology; Enzymes; Genes; Plants (Botany); Yeast; Albumin; Bean Proteins; Hansenula Polymorpha; Saccharomyces Cerevisiae; Theobroma Cacao; Viciline Like Protein; Yeast Expression; Proteins; Recombinant Protein; Article; Cacao; Controlled Study; Gene Expression Regulation; Hansenula Polymorpha; Molecular Cloning; Nonhuman; Priority Journal; Promoter Region; Saccharomyces Cerevisiae; Yeast; Phaseolus (Angiosperm); Pichia Angusta; Saccharomyces Cerevisiae; Theobroma Cacao; Theobroma Cacaoen_US
dc.titleExpression of the major bean proteins from Theobroma cacao (cocoa) in the yeasts Hansenula polymorpha and Saccharomyces cerevisiaeen_US
dc.typeArticleen_US

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