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Isolation and characterization of thermotolerant yeasts for the production of second-generation bioethanol

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Abstract

The purpose of this study was to isolate, identify, and characterize the thermotolerant yeasts for use in high-temperature ethanol fermentation. Thermotolerant yeasts were isolated and screened from soil samples collected from the Mekong Delta, Vietnam, using the enrichment method. Classification and identification of the selected thermotolerant yeasts were performed using matrix-assisted laser desorption ionization/time-of-fight mass spectrometry (MALDI-TOF/MS) and nucleotide sequencing of the D1/D2 domain of the 26S rDNA and the internal transcribed spacer (ITS) 1 and 2 regions. The ethanol production by the selected thermotolerant yeast was carried out using pineapple waste hydrolysate (PWH) as feedstock. A total of 174 yeast isolates were obtained from 80 soil samples collected from 13 provinces in the Mekong Delta, Vietnam. Using MALDI-TOF/MS and nucleotide sequencing of the D1/D2 domain and the ITS 1 and 2 regions, six different yeast species were identified, including Meyerozyma caribbica, Saccharomyces cerevisiae, Candida tropicalis, Torulaspora globosa, Pichia manshurica, and Pichia kudriavzevii. Among the isolated thermotolerant yeasts, P. kudriavzevii CM4.2 displayed great potential for high-temperature ethanol fermentation. The maximum ethanol concentration (36.91 g/L) and volumetric ethanol productivity (4.10 g/L h) produced at 45 °C by P. kudriavzevii CM4.2 were achieved using PWH containing 103.08 g/L of total sugars as a feedstock. These findings clearly demonstrate that the newly isolated thermotolerant yeast P. kudriavzevii CM4.2 has a high potential for second-generation bioethanol production at high temperature.

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Acknowledgments

The authors thank the Faculty of Technology, Khon Kaen University, for providing the Scholarship to Phong HX. A portion of this work was supported by the Graduate School, Khon Kaen University, Ministry of Science and Technology of Vietnam (the project contract no. 09/2014/HĐ-NĐT), Japan Society for the Promotion of Science (JSPS), and the international collaboration research project Core-to-Core Program (CCP). The authors thank Prof. Akio Tani for assistance with the MALDI-TOF/MS analysis.

Funding

This study was funded by the Ministry of Science and Technology of Vietnam (the project contract no. 09/2014/HĐ-NĐT).

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Authors and Affiliations

  1. Department of Biotechnology, Faculty of Technology, Khon Kaen University, Khon Kaen, 40002, Thailand

    Huynh Xuan Phong, Preekamol Klanrit & Pornthap Thanonkeo

  2. Graduate School, Khon Kaen University, Khon Kaen, 40002, Thailand

    Huynh Xuan Phong & Ngo Thi Phuong Dung

  3. Department of Microbial Biotechnology, Biotechnology Research and Development Institute (BiRDi), Can Tho University, Can Tho, 900000, Vietnam

    Huynh Xuan Phong

  4. Fermentation Research Center for Value Added Agricultural Products (FerVAAPs), Khon Kaen University, Khon Kaen, 40002, Thailand

    Preekamol Klanrit & Pornthap Thanonkeo

  5. Department of Biological Chemistry, Faculty of Agriculture, Yamaguchi University, Yamaguchi, 753-8515, Japan

    Mamoru Yamada

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  1. Huynh Xuan Phong
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Correspondence to Pornthap Thanonkeo.

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Phong, H.X., Klanrit, P., Dung, N.T.P. et al. Isolation and characterization of thermotolerant yeasts for the production of second-generation bioethanol. Ann Microbiol 69, 765–776 (2019). https://doi.org/10.1007/s13213-019-01468-5

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