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Construction of Yeast Recombinant (Saccharomyces cerevisiae) having Glucoamylase Activity for Direct Alcohol Fermentation of Starch
Researcher : Purkan
Published : 2008
Group :Other Areas
Full Report : Special request
Abstract :
Insertion of glucoamylase gene in yeast cell of Saccharomyces cerevisiae is required to increase yeast function in ethanol fermentation from starch. Recombinant yeast Saccharomyces cerevisiae which enable producing ethanol from starch in the research had been constructed by amplifying of glucoamylase gene (GLO1), cloning of GLO1 gene in yeast expression vector yEP195-supex2, characterization of recombinant DNA yEP-GLO1, transformation of yeast of S. cerevisiae with recombinant plasmid yEP-GLO1, assaying of recombinant yeast in degradation of starch, and optimizing of bioconversion process of starch into ethanol by  recombinant yeast of Saccharomyces cerevisiae [yEP-GLO1] in the end step. Amplifying of  GLO1 gene of S. fibuligera Rcc64 with FW dan RV primers using PCR technique which was done resulting DNA fragment as much as ± 1.560 bp. Cloning of GLO1 gene which obtained from PCR to vector yEP195-supex2 (± 6.200 bp) had resulted recombinant plasmid yEP-GLO1 (± 7.760 bp). In silico analysis of nucleotides of GLO1 gene in recombinant DNA yEP-GLO1 showed that the gene had 519 residues of amino acid. Transformation of S. cerevisiae with recombinant plasmid yEP-GLO1 could result yeast recombinant having amylolytic activity or enable to digest starch. Starch concentration which could be digested by recombinant yeast of S. cerevisiae [yEP-GLO1] was 10% (w/v). Bioconversion of starch having concentration 10% (b/v) using recombinant yeast  S. cerevisiae BY5207 [yEP-GLO1] could result ethanol as 20% (v/v) to alcoholmeter and 19,5% (v/v) to gas of chromatography. Otherwise, using recombinant yeast  S. cerevisiae S. cerevisiae AS3324 [yEP-GLO1] resulted ethanol as 17% (v/v) to alcoholmeter and 17,5% (v/v) to gas of  chromatography. The highest ethanol in starch bioconversion using both recombinant yeasts BY5207 and AS3324 could be resulted on 144 hours of fermentation time as well as in pH 5.

Key word : Glucoamylase, S. fibuligera Rcc64,   Saccharomyces cereviseae, ethanol fermentation, recombinant yeast