Browse > Article
http://dx.doi.org/10.5352/JLS.2019.29.3.376

Construction of Yeast Strain Suitable for Bioethanol Production by Using Fusion Method  

Kim, Yeon-Hee (Biomedical Engineering and Biotechnology Major, Divison of Applied Bioengineering, Dong-Eui University)
Publication Information
Journal of Life Science / v.29, no.3, 2019 , pp. 376-381 More about this Journal
Abstract
To construct useful yeast strain for bioethanol production, we improved yeast harboring various phenotypes by using yeast protoplast fusion method. In this study, S. cerevisiae BYK-F11 strain which have ethanol tolerance, thermotolerance and ${\beta}-glucanase$ activity and P. $stipitis{\Delta}ura$ strain which has xylose metabolism pathway were fused by genome shuffling. P. $stipitis{\Delta}ura$ strain was constructed for protoplast fusion by URA3 gene disruption, resulting in uracil auxotroph. By protoplast fusion, several fused cells were selected and BYKPS-F8 strain (fused cell) showing both karyotypes from two parent strains (S. cerevisiae BYK-F11 and P. $stipitis{\Delta}ura$ strain) among 22 fused cells was finally selected. Sequentially, various phenotypes such as ${\beta}-glucanase$ activity, xylose utility, ethanol tolerance, thermotolerance and ethanol productivity were analyzed. The BYKPS-F8 strain obtained ${\beta}-glucanase$ activity from BYK-F11 strain and 1.2 fold increased xylose utility from P. $stipitis{\Delta}ura$ strain. Also, the BYKPS-F8 strain showed thermotolerance at $40^{\circ}C$ and increased ethanol tolerance in medium containing 8% ethanol. In this fused cell, 7.5 g/l ethanol from 20 g/l xylose was produced and the multiple phenotypes were stably remained during long term cultivation (260 hr). It was proved that novel biological system (yeast strains) is easily and efficiently bred by protoplast fusion among yeasts having different genus.
Keywords
Ethanol tolerance; karyotype; protoplast fusion; thermotolerance; xylose utility;
Citations & Related Records
Times Cited By KSCI : 1  (Citation Analysis)
연도 인용수 순위
1 Jung H. M. and Kim, Y. H. 2018. Simultaneous Overexpression of Integrated Genes by Copy Number Amplification of a Mini-Yeast Artificial Chromosome. J. Microbiol. Biotechnol. 28, 821-825.   DOI
2 Kim, M. J., Nam, S. W., Tamano, K., Machida, M., Kim, S. K. and Kim, Y. H. 2011. Optimazation for production of exo-${\beta}$-1,3-glucanase (laminarase) from Aspergillus oryzae in Saccharomyces cerevisiae. Kor. Soc. Biotech. Bioeng. 26, 427-432.
3 Park, A. H. and Kim, Y. H. 2013. Breeding of ethanol producing and tolerant Saccharomyces cerevisiae by using genome shuffling. J. Life Sci. 23, 1192-1198.   DOI
4 Sakamoto, T., Hasunuma, T., Hori, Y., Yamada, R. and Kondo, A. 2012. Direct ethanol production from hemicellulosic materials of rice straw by use of an engineered yeast strain codisplaying three types of hemicellulolytic enzymes on the surface of xylose-utilizing Saccharomyces cerevisiae cells. J. Biotechnol. 158, 203-210.   DOI
5 Seok, J. H., Kim, H. S., Hatada, Y., Nam, S. W. and Kim, Y. H. 2012. Construction of an expression system for the secretory production of recombinant ${\alpha}$-agarase in yeast. Biotechnol. Lett. 34, 1041-1049.   DOI
6 Seok, J. H., Park, H. G., Lee, S. H., Nam, S. W., Jeon, S. J., Kim, J. H. and Kim, Y. H. 2010. High-level secretory expression of recombinant ${\beta}$-agarase from Zobellia galactanivarans in Pichia pastoris. Kor. J. Microbiol. Biotechnol. 38, 40-45.
7 Yanagisawa, M., Kawai, S. and Murata, K. 2013. Strategies for the production of high concentrations of bioethanol from seaweeds: production of high concentrations of bioethanol from seaweeds. Bioengineered 4, 224-235.   DOI
8 Sheehan, C. and Weiss, A. S. 1990. Yeast artificial chromosome: rapid extraction for high resolution analysis. Nucleic Acids Res. 18, 2193.   DOI
9 Shi, D. J., Wang, C. L. and Wang, K. M. 2009. Genome shuffling to improve thermotolerance, ethanol tolerance and ethanol productivity of Saccharomyces cerevisiae. J. Mircobiol. Biotechnol. 36, 139-147.
10 Spencer, J. F. T. and Spencer, D. M. 1983. Genetic improvement of industrial yeast. Ann. Rev. Microbiol. 37, 121-142.   DOI
11 Zhu, Y., Wu, L., Zhu, J., Xu, Y. and Yu, S. 2018. Quantitative proteomic analysis of xylose fermentation strain Pichia stipitis CBS 5776 to lignocellulosic inhibitors acetic acid, vanillin and 5-hydroxymethylfurfural. FEMS Microbiol. Lett. 365, doi: 10.1093/femsle/fny245.   DOI
12 Jeon, H. T., Park, U. M. and Kim, K. 2011. The use of aureobasidin A resistant gene as the dominant selectable marker for the selection of industrial yeast hybrid. Kor. J. Microbiol. Biotechnol. 39, 111-118.
13 Attfield, P. V. 1997. Stress tolerance: the key to effective strains of industrial baker's yeast. Nat. Biotechnol. 15, 1351-1357.   DOI
14 Bae, Y. W., Seong, P. J., Cho, D. H., Shin, S. J., Kim, S. W., Han, S. O., Kim, Y. H. and Park, C. H. 2010. Bioethanol Production Based on Lignocellulosic Biomass with Pichia stipitis. KSBBJ 25, 533-538.
15 Harashima, S., Takagi, A. and Oshima, Y. 1984. Transformation of protoplasted yeast cells is directly associated with cell fusion. Mol. Cell Biol. 4, 771-778.   DOI
16 Hou, L. 2010. Improved production of ethanol by novel genome shuffling in Saccharomyces cerevisiae. Appl. Biochem. Biotechnol. 160, 1084-1093.   DOI