Transformant for enhancing bioethanol production, and method for producing ethanol by using said strain
AssigneeKorea University Research and Business Foundation
InventorsSung Ok Han, Kyung Ok Yu, Seung Wook Kim
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Abstract
The present invention relates to a transformant for inhibiting glycerol production through deletion of glycerol producing genes of Saccharomyces cerevisiae modified so as to use glycerol as a fermentation source, or enhancing bioethanol production through overexpression of TATA-binding proteins, SPT3 and SPT15, and a method for producing ethanol by using the transformant.
Claims23
A genetically engineered microbial cell, wherein said cell comprises:
- a disruption of a gene encoding glycerol-3-phosphate dehydrogenase 2 and a disruption of a FPS1 (glycerol facilitator channel) gene encoding yeast glycerol channel FPS1.
A method for preparing a genetically engineered microbial cell which comprises a disruption of a gene encoding a glycerol-3-phosphate dehydrogenase 2 and a disruption of a FPS1 (glycerol facilitator channel) gene encoding yeast glycerol channel FPS1, the method comprising the steps of:
- a) performing PCR using pPICZ vector DNA having a cleavage map of FIG. 8 as a template, a forward primer comprising the sequence from start codon to 40-mer of the gene encoding yeast glycerol channel FPS1 and a reverse primer comprising the sequence from residue 1971 to stop codon of the gene encoding yeast glycerol channel FPS1, thereby obtaining a PCR product comprising the start and stop codons of Fps1 gene;
- b) performing PCR using pET28a vector DNA having a cleavage of FIG. 9 as a template, a forward primer comprising the sequence from start codon to 40-mer of glycerol-3-phosphate dehydrogenase 2 and a reverse primer comprising the sequence from residue 1324 to stop codon of glycerol-3-phosphate dehydrogenase 2, thereby obtaining a PCR product;
- and
- c) performing homologous recombination of the PCR products of steps a) and b) in yeast.
An expression vector pGupSpt3.15 containing a gene encoding Saccharomyces cerevisiae Suppressor of Ty (SPT) 3 and a gene encoding SPT 15.
(canceled)
Dependencies are inferred from the claim text.