Understanding and Efficiently Manipulating Environmental Stress Caused by Metal Ions to Improve Ethanol Fermentation

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Boontiwa Ninchan
Chollada Sirisatesuwon
Kittipong Rattanaporn
Klanarong Sriroth


The inconsistent quality of molasses directly influences ethanol production, particularly due to contamination by metal ions that causes severe problems and reduces production efficiency. This research focused on calcium (Ca2+), potassium (K+), and magnesium (Mg2+) ions that are common in molasses. The key objective was to understand clearly the effect of ions on ethanol fermentation and Saccharomyces cerevisiae performance. Individual ions and ion mixtures were studied in sucrose solution and in molasses. The results showed that severe stress could be ordered as Ca2+>K+>Mg2+, respectively, and the adverse effect was greater when the ion concentration increased. Ca2+ was a strong inhibitor while trace amounts of Mg2+ produced a positive effect. To achieve the greatest efficiency in ethanol production using molasses in the substrate preparation, Ca2+ should not exceed 0.18% (w/w) prior to fermentation and the final sugar concentration should be 20–25% (w/v), as adjusting the addition of sucrose will result in a suitable yeast medium. Pretreatment and dilution were the best practices for ion removal, with Ca2+ being clearly decreased. Furthermore, determination of the composition and ion concentration in molasses is essential to initial steps that must be routinely applied to ensure that the knowledge gained and the efficient techniques investigated can be used to improve ethanol production.

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Ninchan, B., Sirisatesuwon, C., Rattanaporn, K., & Sriroth, K. (2022). Understanding and Efficiently Manipulating Environmental Stress Caused by Metal Ions to Improve Ethanol Fermentation. Applied Science and Engineering Progress, 15(4), 4717. https://doi.org/10.14416/j.asep.2021.06.004
Research Articles


S. Amornvivat, P. Hoontrakul, A. Tamprasirt, L. Larpchevasit, T. Homchampa, S. Khantachavana, and C. Charoenlad, Insight: Bioenergy and the Road to Sustainable Growth. Bangkok, Thailand: Siam Commercial Bank Public Company Limited, 2016.

W. Hicks, “Thailand among ASEAN stars,” 2019. [Online]. Available: https://www.bangkokpost. com/business/1713796/thailand-among-aseanstars

ASEAN UP, “Overview of business in Thailand,” 2017. [Online]. Available: https://aseanup.com/ business-thailand/

N. Tunpaiboon, “Thailand industry outlook 2019–2021: Ethanol industry,” 2019. [Online]. Available: https://www.krungsri.com/bank/get media/0c42d6fd-18d7-41c193696dded234800/ IO_Ethanol_190710_EN_ EX.aspx

Office of Cane and Sugar Board, “Report of ethanol production,” Office of Cane and Sugar Board, Bangkok, Thailand, 2020.

P. Saothong, B. Ninchan, K. Sriroth, K. Rattanaporn, and W. Vanichsriratana, “Kinetics of Saccharomyces cerevisiae fermentation under metal ions stress during ethanol production,” Walailak Journal of Science and Technology (WJST), vol. 18, no. 6, 2021, doi: 10.48048/wjst.2021.9133.

Department of Alternative Energy Development and Efficiency, Ministry of Energy, “Ethanol factories,” 2020. [Online]. Available: https://www.dede.go.th/more_news.php?cid=82 &filename=index

M. A. Clarke, “Syrups,” in Encyclopedia of Food Sciences and Nutrition, 2nd ed. 2003, pp. 5711–5717.

G. N. Abdel-Rahman, N. R. A. Nassar, Y. A. Heikal, M. A. M. Abou-Donia, M. M. Naguib, and M. Fadel, “Effect of different treatments on heavy metal concentration in sugar cane molasses,” International Journal of Agricultural and Biosystems Engineering, vol. 10, no. 1, pp. 43–48, 2016, doi: 10.5281/zenodo. 1125003.

D. Teclua, G. Tivcheva, M. Laingb, and M. Wallis, “Determination of the elemental composition of molasses and its suitability as carbon source for growth of sulphate-reducing bacteria,” Journal of Hazardous Materials, vol. 161, no. 2–3, pp. 1157–1165, 2009, doi: 10.1016/j.jhazmat. 2008.04.120.

Y. Yilmaz, I. Celik, and F. Isik, “Mineral composition and total phenolic content of pomegranate molasses,” Journal of Food, Agriculture & Environment, vol. 5, no. 3–4, pp. 102–104, 2017, doi: 10.1234/ 4.2007.1048.

K. Takeshige and K. Ouchi, “Factors affecting the ethanol productivity of yeast in molasses,” Journal of Fermentation and Bioengineering, vol. 79, no. 5, pp. 449–452, 1995, doi: 10.1016/0922- 338X (95)91260-C.

S. A. Mohamed, A. A. A. Said, A. N. A. Zohri, H. A. Tawfek, and A.A. El-Samman, “Organic and inorganic constituent analysis of cane molasses and its affect on microbial fermentation industries,” in International Conference on: World Perspective for Sugar Crops as Food and Energy Supplies, 2009, pp. 1–16.

S. S. Munsamy, “Investigation into the high ash content in molasses at Nakambala, Zambia,” in the 86th Annual Congress of the South African Sugar Technologists' Association (SASTA 2013), 2013, pp. 67–73.

S. Chotineeranat, R. Wansuksri, K. Piyachomkwan, P. Chatakanonda, P. Weerathaworn, and K. Sriroth, “Effect of calcium ions on ethanol production from molasses by Saccharomyces cerevisiae,” Sugar Tech, vol.12, no. 2, pp. 120–124, 2010.

H. Alexandre, V. Ansanay-Galeote, S. Dequin, and B. Blondin, “Global gene expression during short-term ethanol stress in Saccharomyces cerevisiae,” FEBS Letters, vol. 498, no. 1, pp. 98– 103, 2001, doi:10.1016/s0014-5793(01)02503-0.

J. Bose, O. Babourina, and Z. Rengel, “Role of magnesium in alleviation of aluminium toxicity in plants,” Journal of Experimental Botany, vol. 62, no. 7, pp. 2251–2264, 2011, doi: 10.1093/jxb/ erq456.

C. Sirisatesuwon, B. Ninchan, and K. Sriroth, “Effects of inhibitors on kinetic properties of invertase from Saccharomyces cerevisiae,” Sugar Tech, vol. 22, no. 2, pp. 274–283, 2020, doi: 10.1007/s12355-019-00757-2.

S. Kulshrestha, P. Tyagi, V. Sindhi, and K. S. Yadavilli, “Invertase and its applications – A brie review,” Journal of Pharmacy Research, vol. 7, no. 9, pp. 792–797, 2013, doi:10.1016/j.jopr.2013.07.014.

Vertical Chromatography Co. Ltd., “VertiSepTM sugar HPLC columns,” 2013, [Online]. Available: http://www.vertichrom.com/pdf/hplc_sugar.pdf

Bio-rad Laboratory, “Chromatograpgy: Aminex® HPLC Columns,” 2012. [Online]. Available: https://www.bio-rad.com/webroot/web/pdf/lsr/ literature/Bulletin_6333.pdf

H. O. Udeh and T. E. Kgatla, “Role of magnesium ions on yeast performance during very high gravity fermentation,” Journal of Brewing and Distilling, vol. 4, no. 2, pp. 19–45, 2013, doi:10.5897/JBD 2013.0041.

G. M. Walker, “The roles of magnesium in biotechnology,” Critical Reviews in Biotechnology, vol. 14, no. 4, pp. 311–354, 1994, doi: 10.3109/ 07388559409063643.

G. M. Walker, R. D. Nicola, S. Anthony, and R. Learmonth, “Yeast-metal interactions: impact on brewing and distilling fermentations,” presented at Institute of Brewing & Distilling Asia Pacific Section 2006 Convention, Hobart, Australia, Mar. 19–24, 2006.