Abstract
The ability to grow and produce ethanol under stressful conditions is an important factor in industrial bioethanol production. Trehalose is found in many organisms including Saccharomyces cerevisiae, and has been known to play an important role in enhancing various types of stress tolerance. In this study, Streptomyces albus trehalose-6-phosphate synthase gene (salC) was expressed in Saccharomyces cerevisiae, and the recombinant strain with salC gene showed significantly improved stress resistances and ethanol production. The stress sensitivity and viability tests indicated that the recombinant had a greater resistance to ethanol than the control. At elevated temperatures, the results of flask cultures showed that the expression of salC played a positive role in protecting cells from heat stress. The recombinant strain was found to consume 100 g/L glucose and to produce 39 g/L ethanol at 40°C with an ethanol yield 6% higher than that of the control strain. In the fed-batch experiment in a bioreactor the recombinant strain produced 69 g/L ethanol with about 16% higher yield and about 13% higher productivity than the control strain. This demonstrated the enhancement of ethanol production capabilities of the recombinant strain under a high-ethanol stress condition.
Similar content being viewed by others
References
Mobini-Dehkordi, M., I. Nahvi, H. Zarkesh-Esfahani, K. Ghaedi, M. Tavassoli, and R. Akada (2008) Isolation of a novel mutant strain of Saccharomyces cerevisiae by an ethyl methane sulfonate-induced mutagenesis approach as a high producer of bioethanol. J. Biosci. Bioeng. 105: 403–408.
Kajiwara, S., K. Suga, H. Sone, and K. Nakamura (2000) Improved ethanol tolerance of Saccharomyces cerevisiae strains by increases in fatty acid unsaturation via metabolic engineering. Biotechnol. Lett. 22: 1839–1843.
Singer, M. A. and S. Lindquist (1998) Multiple effects of trehalose on protein folding in vitro and in vivo. Molecular cell. 1: 639–648.
Jain, N. K. and I. Roy (2009) Effect of trehalose on protein structure. Protein Sci. 18: 24–36.
Claudio, V., B. C. Niels, B. Walter, J. E. N. Paul, B. Thomas, and W. Andres (1993) Disruption of TPS2, the gene encoding the 100-kDa subunit of the trehalose-6-phosphate synthase/phosphatase complex in Saccharomyces cerevisiae, causes accumulation of trehalose-6-phosphate and loss of trehalose-6-phosphate phosphatase activity. European J. Biochem. 212: 315–323.
Parrou, J. L., M. -A. Teste, and J. Francois (1997) Effects of various types of stress on the metabolism of reserve carbohydrates in Saccharomyces cerevisiae: Genetic evidence for a stress-induced recycling of glycogen and trehalose. Microbiol. 143: 1891–1900.
Li, L., Y. Ye, L. Pan, Y. Zhu, S. Zheng, and Y. Lin (2009) The induction of trehalose and glycerol in Saccharomyces cerevisiae in response to various stresses. Biochem. Biophysic. Res. Communic. 387: 778–783.
De Virgilio, C., T. Hottiger, J. Dominguez, T. Boller, and A. Wiemken (1994) The role of trehalose synthesis for the acquisition of thermotolerance in yeast. I. Genetic evidence that trehalose is a thermoprotectant. Eur. J. Biochem. 219: 179–186.
Hottiger, T., P. Schmutz, and A. Wiemken (1987) Heat-induced accumulation and futile cycling of trehalose in Saccharomyces cerevisiae. J. Bacteriol. 169: 5518–5522.
Li, H., H. -L. Wang, J. Du, G. Du, J. -C. Zhan, and W. -D. Huang (2010) Trehalose protects wine yeast against oxidation under thermal stress. World J. Microbiol. Biotechnol. 26: 969–976.
Hino, A., K. Mihara, K. Nakashima, and H. Takano (1990) Trehalose levels and survival ratio of freeze-tolerant versus freeze-sensitive yeasts. Appl. Environ. Microbiol. 56: 1386–1391.
Zhang, C. -Y., D. -G. Xiao, and Y. Lv (2010) Influence of trehalose accumulation on response to freeze stress in Baker’s yeast. Proceedings of the Bioinformatics and Biomedical Engineering. June 18–20. Tianjin, China.
Mansure, J. J. C., A. D. Panek, L. M. Crowe, and J. H. Crowe (1994) Trehalose inhibits ethanol effects on intact yeast cells and liposomes. Biochimica et Biophysica Acta (BBA) — Biomembranes. 1191: 309–316.
Sharma, S. C. (1997) A possible role of trehalose in osmotolerance and ethanol tolerance in Saccharomyces cerevisiae. FEMS Microbiol Lett. 152: 11–15.
Ding, J., X. Huang, L. Zhang, N. Zhao, D. Yang, and K. Zhang (2009) Tolerance and stress response to ethanol in the yeast Saccharomyces cerevisiae. Appl.Microbiol. Biotechnol. 85: 253–263.
Arneborg, N., M. K. Moos, and M. Jakobsen (1997) Induction of acetic acid tolerance and trehalose accumulation by added and produced ethanol in Saccharomyces cerevisiae. Biotechnol. Lett. 19: 931–933.
Elbein, A. D., Y. T. Pan, I. Pastuszak, and D. Carroll (2003) New insights on trehalose: a multifunctional molecule. Glycobiol. 13: 17–27.
Choeng, Y. H., J. Y. Yang, G. Delcroix, Y. J. Kim, Y. K. Chang, and S. K. Hong (2007) Expression and characterization of trehalose biosynthetic modules in the adjacent locus of the salbostatin gene cluster. J. Microbiol. Biotechnol. 17: 1675–1681.
Kieser, T., M. J. Bibb, M. J. Buttner, K. F. Chater, and D. A. Hopwood (2000) Practical Streptomyces Genetics. John Innes Foundation, Norwich, UK.
Sambrook, J. and D. W. Russell (2001) Molecular Cloning: A Laboratory Manual. CSHL Press, NY.
Gietz, R. D., R. H. Schiestl, A. R. Willems, and R. A. Woods (1995) Studies on the transformation of intact yeast cells by the LiAc/SS-DNA/PEG procedure. Yeast 11: 355–360.
Lohr, D., P. Venkov, and J. Zlatanova (1995) Transcriptional regulation in the yeast GAL gene family: a complex genetic network. FASEB J. 9: 777–787.
Whang, J., J. Ahn, C. -S. Chun, Y. -J. Son, H. Lee, and E. -S. Choi (2009) Efficient, galactose-free production of Candida antarctica lipase B by GAL10 promoter in Δgal80 mutant of Saccharomyces cerevisiae. Proc. Biochem. 44: 1190–1192.
Vianna, C. R., C. L. Silva, M. J. Neves, and C. A. Rosa (2008) Saccharomyces cerevisiae strains from traditional fermentations of Brazilian cachaca: Trehalose metabolism, heat and ethanol resistance. Antonie Van Leeuwenhoek. 93: 205–217.
Takahashi, T., H. Shimoi, and K. Ito (2001) Identification of genes required for growth under ethanol stress using transposon mutagenesis in Saccharomyces cerevisiae. Mol. Gen. Genom. 265: 1112–1119.
Taylor, M. P., K. L. Eley, S. Martin, M. I. Tuffin, S. G. Burton, and D. A. Cowan (2009) Thermophilic ethanologenesis: Future prospects for second-generation bioethanol production. Trends in Biotechnol. 27: 398–405.
Kiransree, N., M. Sridhar, and L. V. Rao (2000) Characterisation of thermotolerant, ethanol tolerant fermentative Saccharomyces cerevisiae for ethanol production. Bioproc. Biosys. Eng. 22: 243–246.
Balakumar, S., V. Arasaratnam, and K. Balasubramaniam (2001) Isolation and improvement of a thermotolerant Saccharomyces cerevisiae strain. World J. Microbiol. Biotechnol. 17: 739–746.
Nevoigt, E. (2008) Progress in Metabolic Engineering of Saccharomyces cerevisiae. Microbiol. Mol. Biol. Rev. 72: 379–412.
Ansanay-Geleote, V., B. Blondin, S. Dequin, and J. M. Sablayrolles (2001) Stress effect of ethanol on fermentation kinetics by stationary-phase cells of Saccharomyces cerevisiae. Biotechnol. Lett. 23: 677–681.
Patel, P. (2006) Engineered microbes boost ethanol.
Lin, Y. and S. Tanaka (2006) Ethanol fermentation from biomass resources: Current state and prospects. Appl. Microbiol. Biotechnol. 69: 627–642.
Shi, D. -J., C. -L. Wang, and K. -M. Wang (2009) Genome shuffling to improve thermotolerance, ethanol tolerance and ethanol productivity of Saccharomyces cerevisiae. J. Industrial Microbiol. Biotechnol. 36: 139–147.
van Voorst, F., J. Houghton-Larsen, L. Jønson, M. C. Kielland-Brandt, and A. Brandt (2006) Genome-wide identification of genes required for growth of Saccharomyces cerevisiae under ethanol stress. Yeast 23: 351–359.
Wang, Y. J., Y. J. Hao, Z. G. Zhang, T. Chen, J. S. Zhang, and S. Y. Chen (2005) Isolation of trehalose-6-phosphate phosphatase gene from tobacco and its functional analysis in yeast cells. J. Plant Physiol. 162: 215–223.
Bonini, B. M., C. Van Vaeck, C. Larsson, L. Gustafsson, P. Ma, J. Winderickx, P. Van Dijck, and J. M. Thevelein (2000) Expression of Escherichia coli otsA in a Saccharomyces cerevisiae tps1 mutant restores trehalose 6-phosphate levels and partly restores growth and fermentation with glucose and control of glucose influx into glycolysis. Biochem. J. 350: 261–268.
László, V., H. -W. Fehlhaber, and S. Arno (1994) The Trehalase Inhibitor Salbostatin, a Novel Metabolite from Streptomyces albus ATCC21838. Angewandte Chemie International Edition in English 33: 1844–1846.
Author information
Authors and Affiliations
Corresponding authors
Rights and permissions
About this article
Cite this article
Moon, M.H., Ryu, J., Choeng, YH. et al. Enhancement of stress tolerance and ethanol production in Saccharomyces cerevisiae by heterologous expression of a trehalose biosynthetic gene from Streptomyces albus . Biotechnol Bioproc E 17, 986–996 (2012). https://doi.org/10.1007/s12257-012-0148-5
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s12257-012-0148-5