One-step enzymatic hydrolysis of starch using a recombinant strain of Saccharomyces cerevisiae producing α-amylase, glucoamylase and pullulanase

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dc.contributor.authorJanse B.J.H.
dc.contributor.authorPretorius I.S.
dc.date.accessioned2011-05-15T15:56:38Z
dc.date.available2011-05-15T15:56:38Z
dc.date.issued1995
dc.description.abstractA recombinant strain of Saccharomyces cerevisiae was constructed that contained the genes encoding a bacterial α-amylase (AMY1), a yeast glucoamylase (STA2) and a bacterial pullulanase (pulA). The Bacillus amyloliquefaciens α-amylase and S. cerevisiae var. diastaticus glucoamylase genes were expressed in S. cerevisiae using their native promoters and the encoded enzymes secreted under direction of their native leader sequences. In contrast, the Klebsiella pneumoniae pullulanase gene was placed under the control of the Least alcohol dehydrogenase gene promoter (ADC1(P)) and secreted using the yeast mating pheromone α-factor secretion signal (MFα1(S)). Transcription termination of the pullulanase gene was effected by the yeast tryptophan synthase gene terminator (TRP5(T)), whereas termination of the glucoamylase and α-amylase genes was directed by their native terminators. Pullulanase (PUL1) produced by recombinant yeasts containing ADC1(P) MFα1(S) pulA TRP5(T) (designated PUL1) was further characterized and compared to its bacterial counterpart (PulA). The different genes were introduced into S. cerevisiae in different combinations and the various amylolytic Saccharomyces transformants compared to Schwanniomyces occidentalis. Introduction of PUL1 into a S. cerevisiae strain containing both STA2 and AMY1, resulted in 99% assimilation of starch.
dc.description.versionArticle
dc.identifier.citationApplied Microbiology and Biotechnology
dc.identifier.citation42
dc.identifier.citation6
dc.identifier.issn1757598
dc.identifier.other10.1007/s002530050346
dc.identifier.urihttp://hdl.handle.net/10019.1/9955
dc.subjectamylase
dc.subjectglucan 1,4 alpha glucosidase
dc.subjectpullulanase
dc.subjectstarch
dc.subjectarticle
dc.subjectbiodegradation
dc.subjectgene control
dc.subjectgene expression
dc.subjecthydrolysis
dc.subjectmolecular cloning
dc.subjectnonhuman
dc.subjectsaccharomyces cerevisiae
dc.subjectalpha-Amylase
dc.subjectBacillus
dc.subjectGene Expression Regulation, Fungal
dc.subjectGenes, Bacterial
dc.subjectGenes, Fungal
dc.subjectGlucan 1,4-alpha-Glucosidase
dc.subjectGlycoside Hydrolases
dc.subjectHydrolysis
dc.subjectKlebsiella pneumoniae
dc.subjectPlasmids
dc.subjectRecombination, Genetic
dc.subjectRestriction Mapping
dc.subjectSaccharomyces cerevisiae
dc.subjectStarch
dc.titleOne-step enzymatic hydrolysis of starch using a recombinant strain of Saccharomyces cerevisiae producing α-amylase, glucoamylase and pullulanase
dc.typeArticle
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