Cited by 215Fungal and yeast genetics researchBiofuel production and bioconversionMicrobial Metabolic Engineering and Bioproduction

High-Temperature Ethanol Fermentation and Transformation with Linear DNA in the Thermotolerant Yeast Kluyveromyces marxianus DMKU3-1042

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Nonklang, Sanom, Abdel-Banat, Babiker M. A., Cha-aim, Kamonchai, Moonjai, Nareerat, Hoshida, Hisashi, Limtong, Savitree, Yamada, Mamoru, Akada, Rinji·2008·Applied and Environmental Microbiology

Abstract

We demonstrate herein the ability of Kluyveromyces marxianus to be an efficient ethanol producer and host for expressing heterologous proteins as an alternative to Saccharomyces cerevisiae . Growth and ethanol production by strains of K. marxianus and S. cerevisiae were compared under the same conditions. K. marxianus DMKU3-1042 was found to be the most suitable strain for high-temperature growth and ethanol production at 45°C. This strain, but not S. cerevisiae , utilized cellobiose, xylose, xylitol, arabinose, glycerol, and lactose. To develop a K. marxianus DMKU3-1042 derivative strain suitable for genetic engineering, a uracil auxotroph was isolated and transformed with a linear DNA of the S. cerevisiae Sc URA3 gene. Surprisingly, Ura + transformants were easily obtained. By Southern blot hybridization, the linear Sc URA3 DNA was found to have inserted randomly into the K. marxianus genome. Sequencing of one Lys − transformant confirmed the disruption of the Km LYS1 gene by the Sc URA3 insertion. A PCR-amplified linear DNA lacking K. marxianus sequences but containing an Aspergillus α-amylase gene under the control of the Sc TDH3 promoter together with an Sc URA3 marker was subsequently used to transform K. marxianus DMKU3-1042 in order to obtain transformants expressing Aspergillus α-amylase. Our results demonstrate that K. marxianus DMKU3-1042 can be an alternative cost-effective bioethanol producer and a host for transformation with linear DNA by use of S. cerevisiae -based molecular genetic tools.

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  1. [5]Banat, I. M., P. Nigam, D. Singh, R. Marchantand, and A. P. McHale. 1998. Ethanol production at elevated temperatures and alcohol concentrations. I. Yeasts in general. World J. Microbiol. Biotechnol. 14:809-821. [Google Scholar]
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  3. [42]Singh, D., P. Nigam, I. M. Banat, I. M. Manta, R. Marchant, and A. P. McHale. 1998. Ethanol production at elevated temperatures and alcohol concentrations. II. Use of Kluyveromyces marxianus IBM3. World J. Microbiol. Biotechnol. 14:823-834. [Google Scholar]

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Authors
Nonklang, S., Abdel-Banat, B. M. A., Cha-aim, K., Moonjai, N., Hoshida, H., Limtong, S., Yamada, M., & Akada, R.
Affiliations
  • Yamaguchi University (JP)
  • Department of Applied Molecular Bioscience, Yamaguchi University Graduate School of Medicine, Tokiwadai, Ube 755-8611, Japan
  • Ratchathani University (TH)
  • Kasetsart University (TH)
Corresponding author
Akada, Rinji
Venue
Applied and Environmental Microbiology
Published
2008-12-15
Volume
74
Issue
24
Pages
7514-7521
ISSN
  • 0099-2240 (ISSN-L)
  • 1098-5336
OpenAlex
W2116207915
Cited by
215
References
50
Citations by year
2026: 4 · 2025: 11 · 2024: 13 · 2023: 10 · 2022: 9 · 2021: 15 · 2020: 11 · 2019: 14 · 2018: 17 · 2017: 18 · 2016: 7 · 2015: 12 · 2014: 20 · 2013: 12 · 2012: 16
FWCI
3.42
Type
journal article
Language
en
Open access
Locations
  • Applied and Environmental Microbiology (publishedVersion)
  • Europe PMC (PubMed Central) (publishedVersion)
  • PubMed Central (publishedVersion)
  • PubMed (publishedVersion)
  • Institutional Repositories DataBase (IRDB) (submittedVersion)
Funders
  • National Research Council of Thailand
  • Japan Society for the Promotion of Science
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Fungal and yeast genetics research
  • Physical Sciences › Engineering › Biomedical Engineering › Biofuel production and bioconversion
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Microbial Metabolic Engineering and Bioproduction
Keywords
Kluyveromyces marxianus, heterologous expression, Saccharomyces cerevisiae comparison, xylose utilization, bioethanol production, thermotolerant yeast
MeSH terms
Ethanol, alpha-Amylases, Base Sequence, DNA, Fungal, Fermentation, Fungal Proteins, Hot Temperature, Kluyveromyces, Molecular Sequence Data, Recombination, Genetic, Saccharomyces cerevisiae, Transformation, Genetic, Blotting, Southern, Sequence Alignment, Carbohydrate Metabolism

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