Antonie van Leeuwenhoek

, Volume 112, Issue 7, pp 975–990 | Cite as

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

  • Sornsiri Pattanakittivorakul
  • Noppon Lertwattanasakul
  • Mamoru Yamada
  • Savitree LimtongEmail author
Original Paper


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.


Ethanol fermentation Thermotolerant yeast Molasses Mutagenesis Ethyl methane sulfonate 


Authors 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.


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.

Ethical approval

This article does not contain any studies with human participants or animals performed by any of the authors.


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Copyright information

© Springer Nature Switzerland AG 2019

Authors and Affiliations

  • Sornsiri Pattanakittivorakul
    • 1
  • Noppon Lertwattanasakul
    • 1
  • Mamoru Yamada
    • 2
  • Savitree Limtong
    • 1
    • 3
    Email author
  1. 1.Department of Microbiology, Faculty of ScienceKasetsart UniversityChatuchak, BangkokThailand
  2. 2.Applied Molecular Bioscience, Graduate School of MedicineYamaguchi UniversityUbeJapan
  3. 3.Academy of ScienceThe Royal Society of ThailandBangkokThailand

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