High-Temperature Ethanol Fermentation and Transformation with Linear DNA in the Thermotolerant Yeast Kluyveromyces marxianus DMKU3-1042
Open accessAbstract
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.
The claims and statistics of this paper haven’t been checked yet.
Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.
Integrity signals
Registry
Cites
Statistics
Claims the paper makes
The claims and statistics of this paper haven’t been checked yet.
Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.
Claims other papers make about this one
Each is checked against this paper’s own text: does it say what the citing paper says it does?
Several types of microorganisms such as bacteria, yeasts, and fungi have been identified for their potential to ferment sugars to bioethanol.
“Several types of microorganisms, such as bacteria, yeasts, and fungi, have been identified for their potential for fermenting sugars to bioethanol [123–125].”
From The promising future of microalgae: Current status, challenges, and optimization of a sustainable and renewable industry for biofuels, feed, and other products (2018) · cites this paper as [124]
Self-check
Percentages, sums and reported differences re-computed from the paper’s own numbers.
The self-check needs the paper’s full text.
Cites
Molecular Cloning: A Laboratory Manual
Sambrook, J., Fritsch, E., & Maniatis, T. (1989) · UCLA - Biblioteca de Ciencias de la Salud · Cited by 133,589 · see citing works
Molecular Cloning. A Laboratory Manual
Wood, E. (1983) · Biochemical Education · doi:10.1016/0307-4412(83)90068-7 · Cited by 29,534 · see citing works
Sikorski, R. S., & Hieter, P. (1989) · Genetics · doi:10.1093/genetics/122.1.19 · Cited by 8,685 · see citing works
Sherman, F., Fink, G. R., & Hicks, J. B. (1979) · Cold Spring Harbor Laboratory eBooks · Cited by 7,543 · see citing works
Brachmann, C. B., Davies, A., Cost, G. J., Caputo, E. M., Li, J., Hieter, P., & Boeke, J. D. (1998) · Yeast · doi:10.1002/(sici)1097-0061(19980130)14:2 · Cited by 3,411 · see citing works
Transformation of yeast by lithium acetate/single-stranded carrier DNA/polyethylene glycol method
Gietz, R. D., & Woods, R. A. (2002) · Methods in enzymology on CD-ROM/Methods in enzymology · doi:10.1016/s0076-6879(02)50957-5 · Cited by 2,890 · see citing works
Dujon, B., Sherman, D., Fischer, G., Durrens, P., Casaregola, S., Lafontaine, I., de Montigny, J., Marck, C., Neuvéglise, C., Talla, E., et al. (2004) · Nature · doi:10.1038/nature02579 · Cited by 1,623 · see citing works
Liu, Y., Mitsukawa, N., Oosumi, T., & Whittier, R. F. (1995) · The Plant Journal · doi:10.1046/j.1365-313x.1995.08030457.x · Cited by 1,387 · see citing works
[10] 5-Fluoroorotic acid as a selective agent in yeast molecular genetics
Boeke, J. D., Trueheart, J., Natsoulis, G., & Fink, G. R. (1987) · Methods in enzymology on CD-ROM/Methods in enzymology · doi:10.1016/0076-6879(87)54076-9 · Cited by 1,368 · see citing works
The yeast Kluyveromyces marxianus and its biotechnological potential
Fonseca, G. G., Heinzle, E., Wittmann, C., & Gombert, A. K. (2008) · Applied Microbiology and Biotechnology · doi:10.1007/s00253-008-1458-6 · Cited by 563 · see citing works
The yeast genome project: what did we learn?
Dujon, B. (1996) · Trends in Genetics · doi:10.1016/0168-9525(96)10027-5 · Cited by 551 · see citing works
Large-scale analysis of the yeast genome by transposon tagging and gene disruption
Ross‐Macdonald, P., Coelho, P. S. R., Roemer, T., Agarwal, S., Kumar, A., Jansen, R., Cheung, K., Sheehan, A. E., Symoniatis, D., Umansky, L., et al. (1999) · Nature · doi:10.1038/46558 · Cited by 530 · see citing works
Limtong, S., Sringiew, C., & Yongmanitchai, W. (2007) · Bioresource Technology · doi:10.1016/j.biortech.2006.10.044 · Cited by 287 · see citing works
Self-citationMetabolic Engineering for Pentose Utilization in Saccharomyces cerevisiae
Hahn‐Hägerdal, B., Karhumaa, K., Jeppsson, M., & Gorwa‐Grauslund, M. F. (2007) · Springer eBooks · doi:10.1007/10_2007_062 · Cited by 280 · see citing works
A Modified Saccharomyces cerevisiae Strain That Consumes l -Arabinose and Produces Ethanol
Becker, J., & Boles, E. (2003) · Applied and Environmental Microbiology · doi:10.1128/aem.69.7.4144-4150.2003 · Cited by 253 · see citing works
van Maris, A. J. A., Winkler, A. A., Kuyper, M., de Laat, W. T. A. M., van Dijken, J. P., & Pronk, J. T. (2007) · Advances in biochemical engineering, biotechnology · doi:10.1007/10_2007_057 · Cited by 245 · see citing works
Banat, I. M., Nigam, P. S. N., & Marchant, R. (1992) · World Journal of Microbiology and Biotechnology · doi:10.1007/bf01201874 · Cited by 223 · see citing works
Wisselink, H. W., Toirkens, M. J., del Rosario Franco Berriel, M., Winkler, A. A., van Dijken, J. P., Pronk, J. T., & van Maris, A. J. A. (2007) · Applied and Environmental Microbiology · doi:10.1128/aem.00177-07 · Cited by 209 · see citing works
Ethanol production from crude whey by Kluyveromyces marxianus
Zafar, S., & Owais, M. (2005) · Biochemical Engineering Journal · doi:10.1016/j.bej.2005.05.009 · Cited by 156 · see citing works
Construction of thermotolerant yeast expressing thermostable cellulase genes
Hong, J., Wang, Y., Kumagai, H., et al. (2007) · Journal of Biotechnology · doi:10.1016/j.jbiotec.2007.03.008 · Cited by 150 · see citing works
PCR‐mediated seamless gene deletion and marker recycling in Saccharomyces cerevisiae
Akada, R., Kitagawa, T., Kaneko, S., Toyonaga, D., Ito, S., Kakihara, Y., Hoshida, H., Morimura, S., Kondo, A., & Kida, K. (2006) · Yeast · doi:10.1002/yea.1365 · Cited by 123 · see citing works
Self-citationDirect Fermentation of d -Xylose to Ethanol by Kluyveromyces marxianus Strains
Margaritis, A., & Bajpai, P. (1982) · Applied and Environmental Microbiology · doi:10.1128/aem.44.5.1039-1041.1982 · Cited by 110 · see citing works
Cloning and sequencing of the inulinase gene of Kluyveromyces marxianus var. marxianus ATCC 12424
Laloux, O., Cassart, J.-P., Delcour, J., et al. (1991) · FEBS Letters · doi:10.1016/0014-5793(91)80909-m · Cited by 96 · see citing works
Isolation of Auxotrophic Mutants of Diploid Industrial Yeast Strains after UV Mutagenesis
Hashimoto, S., Ogura, M., Aritomi, K., et al. (2005) · Applied and Environmental Microbiology · doi:10.1128/aem.71.1.312-319.2005 · Cited by 88 · see citing works
Anderson, P. J., McNeil, K., & Watson, K. (1986) · Applied and Environmental Microbiology · doi:10.1128/aem.51.6.1314-1320.1986 · Cited by 84 · see citing works
Hughes, D. B., Tudroszen, N. J., & Moye, C. J. (1984) · Biotechnology Letters · doi:10.1007/bf00128221 · Cited by 71 · see citing works
Sedlák, M., & Ho, N. W. Y. (2001) · Enzyme and Microbial Technology · doi:10.1016/s0141-0229(00)00282-9 · Cited by 64 · see citing works
Steensma, H. Y., Jongh, F. C. M. D., & Linnekamp, M. (1988) · Current Genetics · doi:10.1007/bf00419987 · Cited by 64 · see citing works
Large-Scale Mutagenesis of the Yeast Genome Using a Tn7-Derived Multipurpose TransposonPreprint
Kumar, A., Seringhaus, M. R., Biery, M. C., et al. (2004) · Genome Research · doi:10.1101/gr.2875304 · Cited by 61 · see citing works
Application of the Cre-loxP system for multiple gene disruption in the yeast Kluyveromyces marxianus
Ribeiro, O., Gombert, A. K., Teixeira, J. A., & Domingues, L. (2007) · Journal of Biotechnology · doi:10.1016/j.jbiotec.2007.05.027 · Cited by 55 · see citing works
Wilkins, M. R., Mueller, M., Eichling, S., et al. (2008) · Process Biochemistry · doi:10.1016/j.procbio.2007.12.011 · Cited by 52 · see citing works
Sequential gene integration for the engineering of Kluyveromyces marxianus
Pecota, D. C., Rajgarhia, V. B., & Silva, N. A. D. (2006) · Journal of Biotechnology · doi:10.1016/j.jbiotec.2006.07.031 · Cited by 50 · see citing works
Rajoka, M. I., Ferhan, M., & Khalid, A. M. (2005) · Letters in Applied Microbiology · doi:10.1111/j.1472-765x.2005.01663.x · Cited by 49 · see citing works
Purification and properties of the β‐fructofuranosidase from Kluyveromyces fragilis
Workman, W. E., & Day, D. F. (1983) · FEBS Letters · doi:10.1016/0014-5793(83)80927-2 · Cited by 42 · see citing works
Shimoda, C., Itadani, A., Sugino, A., et al. (2006) · Genes & Genetic Systems · doi:10.1266/ggs.81.391 · Cited by 41 · see citing works
Comparison of yeast strains for batch ethanol fermentation of cheese?whey powder (CWP) solution
Özmıhçı, S., & Kargı, F. (2007) · Letters in Applied Microbiology · doi:10.1111/j.1472-765x.2007.02132.x · Cited by 39 · see citing works
Yeasts Occurring in Souring Figs
Mrak, E. M., Phaff, H. J., Vaughn, R. H., et al. (1942) · Journal of Bacteriology · doi:10.1128/jb.44.4.441-450.1942 · Cited by 38 · see citing works
Akada, R., Shimizu, Y., Matsushita, Y., et al. (2001) · Yeast · doi:10.1002/yea.797 · Cited by 35 · see citing works
Self-citationIborra, F., & Ball, M. M. (1994) · Yeast · doi:10.1002/yea.320101211 · Cited by 34 · see citing works
Utilization of corn silage juice by Klyuveromyces marxianus
Hang, Y. D., Woodams, E. E., & Hang, L. E. (2002) · Bioresource Technology · doi:10.1016/s0960-8524(02)00170-0 · Cited by 31 · see citing works
Lertwattanasakul, N., Sootsuwan, K., Limtong, S., et al. (2007) · Bioscience Biotechnology and Biochemistry · doi:10.1271/bbb.60622 · Cited by 31 · see citing works
Self-citationBartkevic̆iūt≐, D., S̆ieks̆tel≐, R., & Sasnauskas, K. (2000) · Enzyme and Microbial Technology · doi:10.1016/s0141-0229(00)00155-1 · Cited by 30 · see citing works
High efficiency transformation of Kluyveromyces marxianus by a replicative plasmid
Iborra, F. (1993) · Current Genetics · doi:10.1007/bf00324685 · Cited by 28 · see citing works
Dellaporta, S. L., & Moreno, M. A. (1994) · Springer eBooks · doi:10.1007/978-1-4612-2694-9_99 · Cited by 26 · see citing works
Martini, A., Phaff, H. J., & Douglass, S. A. (1972) · Journal of Bacteriology · doi:10.1128/jb.111.2.481-487.1972 · Cited by 21 · see citing works
Application of a microtitre reader system to the screening of inulinase nulinase-producing yeasts
Passador-Gurgel, G. C., Furlan, S. Â., Melier, J. K., et al. (1996) · Applied Microbiology and Biotechnology · doi:10.1007/s002530050664 · Cited by 16 · see citing works
Ball, M. M., Raynal, A., Guérineau, M., et al. (1999) · PubMed · Cited by 16 · see citing works
Martini, A. V., Rosini, G., & Martini, A. V. (1988) · Biotechnology Techniques · doi:10.1007/bf01875545 · Cited by 14 · see citing works
Effect of pH on the growth of Kluyveromyces fragilis on deproteinized whey.
de Sánchez, S. B., & Castillo, F. J. (1980) · PubMed · Cited by 7 · see citing works
Faculty and students: academic and disciplinary decisions.
Northrop, C. (1984) · PubMed · Cited by 1 · see citing works
Flagged first, then by citations
Research works printed but not linked: 3
- [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]
- [23]Kaiser, C., S. Michaelis, and A. Mitchell. 1994. Methods in yeast genetics. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY.
- [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]
Cited by
Sorted by citations
Khan, M. I., Shin, J. H., & Kim, J. D. (2018) · Microbial Cell Factories · doi:10.1186/s12934-018-0879-x · Cited by 2,082 · see citing works
Comparative Review on the Production and Purification of Bioethanol from Biomass: A Focus on Corn
Assaf, J. C., Mortada, Z., Rezzoug, S.-A., Maache-Rezzoug, Z., Debs, E., & Louka, N. (2024) · Processes · doi:10.3390/pr12051001 · Cited by 51 · see citing works
- 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
- Publisher
- American Society for Microbiology
- Published
- 2008-12-15
- Volume
- 74
- Issue
- 24
- Pages
- 7514-7521
- ISSN
- 0099-2240 (ISSN-L)
- 1098-5336
- PMC
- PMC2607150
- 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
- green
- www.ncbi.nlm.nih.gov
- 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
About these checks
Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.
Statistics and arithmetic are recomputed, not judged.
Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.
What counts as a claim ›How we measure confidence ›How we check this ›
Are you an author, or do you see a problem with this page? Request a review