Selection of thermotolerant Saccharomyces cerevisiae for high temperature ethanol production from molasses and increasing ethanol production by strain improvement
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A thermotolerant ethanol fermenting yeast strain is a key requirement for effective ethanol production at high temperature. This work aimed to select a thermotolerant yeast producing a high ethanol concentration from molasses and increasing its ethanol production by mutagenesis. Saccharomyces cerevisiae DMKU 3-S087 was selected from 168 ethanol producing strains because it produced the highest ethanol concentration from molasses at 40 °C. Optimization of molasses broth composition was performed by the response surface method using Box–Behnken design. In molasses broth containing optimal total fermentable sugars (TFS) of 200 g/L and optimal (NH4)2SO4 of 1 g/L, with an initial pH of 5.5 by shaking flask cultivation at 40 °C ethanol, productivity and yield were 58.4 ± 0.24 g/L, 1.39 g/L/h and 0.29 g/g, respectively. Batch fermentation in a 5 L stirred-tank fermenter with 3 L optimized molasses broth adjusted to an initial pH of 5.5 and fermentation controlled at 40 °C and 300 rpm agitation resulted in 72.4 g/L ethanol, 1.21 g/L/h productivity and 0.36 g/g yield at 60 h. Strain DMKU 3-S087 improvement was performed by mutagenesis using ultraviolet radiation and ethyl methane sulfonate (EMS). Six EMS mutants produced higher ethanol (65.2 ± 0.48–73.0 ± 0.54 g/L) in molasses broth containing 200 g/L TFS and 1 g/L (NH4)2SO4 by shake flask fermentation at 37 °C than the wild type (59.8 ± 0.25 g/L). Among these mutants, only mutant S087E100-265 produced higher ethanol (62.5 ± 0.26 g/L) than the wild type (59.5 ± 0.02 g/L) at 40 °C. In addition, mutant S087E100-265 showed better tolerance to high sugar concentration, furfural, hydroxymethylfurfural and acetic acid than the wild type.
KeywordsEthanol fermentation Thermotolerant yeast Molasses Mutagenesis Ethyl methane sulfonate
SP: Performed research and wrote the paper, NL: Data discussion and checking the paper, MY: Data discussion and checking the paper, SL: Designed study, data discussion and wrote the paper.
This work was supported by the research grant from the Faculty of Science, Kasetsart University, Bangkok, Thailand and the Thailand Research Fund through the TRF Research-Team Promotion Grant RTA6080004.
Compliance with ethical standards
Conflict of interest
The authors declare that there are no conflicts of interest.
This article does not contain any studies with human participants or animals performed by any of the authors.
- Brooks AA (2008) Ethanol production potential of local yeast strains isolated from ripe banana peels. Afr J Biotechnol 7:3749–3752Google Scholar
- Gasmalla MAA, Yang R, Nikoo M, Man S (2012) Production of ethanol from Sudanese sugar cane molasses and evaluation of its quality. J Food Process Technol 3:1–3Google Scholar
- Hamouda HI, Nassar HN, Madian HR, Abu Amr SS, EI-Gendy NS (2015a) Statistical optimization of batch ethanol fermentation of sugarcane molasses by Candida tropicalis strain HSC-24. Int J Chemtech Res 8:878–889Google Scholar
- Hughes SR, Gibbons WR, Bang SS, Pinkelman R, Bischoff KM, Slininger PJ, Qureshi N, Kurtzman CP, Liu SQ, Saha BC, Jackson JS, Catta MA, Rich JO, Javers JE (2012) Random UV-C mutagenesis of Scheffersomyces (formerly Pichia) stipitis NRRL Y-7124 to improve anaerobic growth on lignocellulosic sugars. J Ind Microbiol Biotechnol 39:163–173CrossRefGoogle Scholar
- Taherzadeh MJ, Karimi K (2011) Fermentation inhibitors in ethanol processes and different strategies to reduce their effects. In: Pandey A, Larroche C, Ricke SC, Dussap CG, Gnansounou E (eds) Receptor localization. Elsevier, Burlington, pp 287–311Google Scholar
- Watanabe T, Srichuwong S, Arakane M, Tamiya S, Yoshinaga M, Watanabe I, Yamamoto M, Ando A, Tokuyasu K, Nakamura T (2010) Selection of stress-tolerant yeasts for simultaneous saccharification and fermentation (SSF) of very high gravity (VHG) potato mash to ethanol. Bioresour Technol 101:9710–9714CrossRefGoogle Scholar