Direct and Efficient Production of Ethanol from Cellulosic Material with a Yeast Strain Displaying Cellulolytic Enzymes
0106 biological sciences
2. Zero hunger
Ethanol
Recombinant Fusion Proteins
Genetic Vectors
Saccharomyces cerevisiae
Enzymes, Immobilized
01 natural sciences
Peptide Fragments
Culture Media
Cellulase
Microscopy, Fluorescence
Fermentation
Genes, Synthetic
Cellulose
Genetic Engineering
Chromatography, High Pressure Liquid
DOI:
10.1128/aem.68.10.5136-5141.2002
Publication Date:
2002-09-25T18:02:42Z
AUTHORS (10)
ABSTRACT
ABSTRACT
For direct and efficient ethanol production from cellulosic materials, we constructed a novel cellulose-degrading yeast strain by genetically codisplaying two cellulolytic enzymes on the cell surface of
Saccharomyces cerevisiae
. By using a cell surface engineering system based on α-agglutinin, endoglucanase II (EGII) from the filamentous fungus
Trichoderma reesei
QM9414 was displayed on the cell surface as a fusion protein containing an RGSHis6 (Arg-Gly-Ser-His
6
) peptide tag in the N-terminal region. EGII activity was detected in the cell pellet fraction but not in the culture supernatant. Localization of the RGSHis6-EGII-α-agglutinin fusion protein on the cell surface was confirmed by immunofluorescence microscopy. The yeast strain displaying EGII showed significantly elevated hydrolytic activity toward barley β-glucan, a linear polysaccharide composed of an average of 1,200 glucose residues. In a further step, EGII and β-glucosidase 1 from
Aspergillus aculeatus
No. F-50 were codisplayed on the cell surface. The resulting yeast cells could grow in synthetic medium containing β-glucan as the sole carbon source and could directly ferment 45 g of β-glucan per liter to produce 16.5 g of ethanol per liter within about 50 h. The yield in terms of grams of ethanol produced per gram of carbohydrate utilized was 0.48 g/g, which corresponds to 93.3% of the theoretical yield. This result indicates that efficient simultaneous saccharification and fermentation of cellulose to ethanol are carried out by a recombinant yeast cells displaying cellulolytic enzymes.
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