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Selection of thermotolerant Saccharomyces cerevisiae for high temperature ethanol production from molasses and increasing ethanol production by strain improvement

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Abstract

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.

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Funding

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.

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Authors and Affiliations

  1. Department of Microbiology, Faculty of Science, Kasetsart University, Chatuchak, Bangkok, 10900, Thailand

    Sornsiri Pattanakittivorakul, Noppon Lertwattanasakul & Savitree Limtong

  2. Applied Molecular Bioscience, Graduate School of Medicine, Yamaguchi University, Ube, Yamaguchi, 755-8505, Japan

    Mamoru Yamada

  3. Academy of Science, The Royal Society of Thailand, Bangkok, 10300, Thailand

    Savitree Limtong

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  1. Sornsiri Pattanakittivorakul
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  2. Noppon Lertwattanasakul
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  3. Mamoru Yamada
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  4. Savitree Limtong
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Contributions

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.

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Correspondence to Savitree Limtong.

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Pattanakittivorakul, S., Lertwattanasakul, N., Yamada, M. et al. Selection of thermotolerant Saccharomyces cerevisiae for high temperature ethanol production from molasses and increasing ethanol production by strain improvement. Antonie van Leeuwenhoek 112, 975–990 (2019). https://doi.org/10.1007/s10482-019-01230-6

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