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Research on Production of Cellulosic Ethanol – Development of High-Efficiency Yeast Strain using Mutagenesis and Selection and Self-Cloning, and Study of Entire Process –

Article of Honda R&D Technical Review Vol.28 No.2


A fermentation organism capable of producing a high fermentation yield in the presence of inhibitors, necessary for the realization of increased efficiency in the process of production of second-generation bioethanol from inedible plant biomass, has been developed. The construction of the organism involved a combination of mutagenesis and selection and self-cloning, and results indicated the effectiveness of this combination in the creation of fermentation organisms. Self-cloning is a form of genetic modification implemented within a scope that makes it possible to treat the organism as non-genetically-modified.
Mutagenesis and selection using biomass hydrolysates increased both the organism’s tolerance to inhibitors in hydrolysates and its ethanol fermentation yield. Using the selected strain as a base, genes for the expression of transaldolase and alcohol dehydrogenase were enhanced by self-cloning. The developed strain was able to produce a higher concentration of ethanol.
An examination of the entire process led to the discovery of a biomass pre-treatment condition that would control the production of phenols, which act as inhibitors. A process using this pre-treatment condition and the developed fermentation organism would be able to produce 213 L of ethanol from 1 dry-ton of corn stover.


(1) Liu, Z. L.: Molecular mechanisms of yeast tolerance and in situ detoxification of lignocellulose hydrolysates, Applied Microbiology and Biotechnology, Vol. 90, p. 809-825, (2011)
(2) http://faostat.fao.org/site/728/DesktopDefault.aspx?PageID=728#ancor
(3) Hasunuma, T., Sanda, T., Yamada, R., Yoshimura, K., Ishii, J., Kondo, A.: Metabolic pathway engineering based on metabolomics confers acetic and formic acid tolerance to a recombinant xylose-fermenting strain of Saccharomyces cerevisiae, Microbial Cell Factories, Vol. 10, p. 2-13, (2011)
(4) www.yeastgenome.org
(5) Foureau, E., Courdavault, V., Simkin, A. J., Pichon, O., Crèche, J., Giglioli-Guivarc’h, N., Clastre, M., Papon, N.: Optimization of the URA-blaster disruption system in Candida guilliermondii: efficient gene targeting using the URA3 marker, Journal of Microbiological Methods, Vol. 91, p. 117-120, (2012)
(6) Process Design and Economics for Biochemical Conversion of Lignocellulosic Biomass to Ethanol, http://www.nrel.gov/docs/fy11osti/47764.pdf, NREL, May 2011

Author (organization or company)

Yoshiki TSUCHIDA(Fundamental Technology Research Center)、Norihiko TSUKAGOSHI(Fundamental Technology Research Center)

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