Construction of flocculent Kluyveromyces marxianus strains suitable for high-temperature ethanol fermentation

Flocculating yeasts are highly useful in fermentation processes because these cells can be separated easily from the fermentation mash. However, native yeasts are usually non-flocculating, including Kluyveromyces marxianus, which exhibits a potent high-temperature ethanol fermentation ability. We de...

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Bibliographic Details
Published in:Bioscience, biotechnology, and biochemistry biotechnology, and biochemistry, 2009-05, Vol.73 (5), p.1090-1095
Main Authors: Nonklang, S.(Yamaguchi Univ. (Japan)), Ano, A, Abdel-Banat, B.M.A, Saito, Y, Hoshida, H, Akada, R
Format: Article
Language:English
Subjects:
ALCOHOLIC FERMENTATION
Biological and medical sciences
Biotechnology
CALOR
CHALEUR
Ethanol - metabolism
ethanol fermentation
FERMENTACION ALCOHOLICA
Fermentation
FERMENTATION ALCOOLIQUE
FLOCCULATION
FLOCULACION
FLOCULATION
Fundamental and applied biological sciences. Psychology
Genetic Engineering - methods
GENETIC TRANSFORMATION
HEAT
Hot Temperature
Kluyveromyces - genetics
Kluyveromyces - metabolism
KLUYVEROMYCES MARXIANUS
Methods. Procedures. Technologies
Microbial engineering. Fermentation and microbial culture technology
Molasses
SACCHAROMYCES CEREVISIAE
Saccharomyces cerevisiae - genetics
Saccharomyces cerevisiae Proteins - genetics
TRANSFORMACION GENETICA
TRANSFORMATION GENETIQUE
yeast
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Summary:Flocculating yeasts are highly useful in fermentation processes because these cells can be separated easily from the fermentation mash. However, native yeasts are usually non-flocculating, including Kluyveromyces marxianus, which exhibits a potent high-temperature ethanol fermentation ability. We describe here the construction of flocculent K. marxianus strains via the introduction of the FLO1, FLO5, FLO9, and FLO10 genes from Saccharomyces cerevisiae. The S. cerevisiae FLO genes were overexpressed by upstream insertion of the constitutive TDH3 promoter, resulting in flocculent S. cerevisiae strains. These TDH3p-FLO sequences were then amplified by PCR and introduced directly into a K. marxianus strain. These K. marxianus strains showed a flocculation phenotype, indicating that the introduced S. cerevisiae TDH3 promoter and all FLO genes were functional in this strain. Moreover, a flocculating K. marxianus strain showed the same ethanol production profile as that of its wild-type parent. The K. marxianus flocculating strains we generated should be useful in the future development of cost-effective fed-batch and continuous fermentation systems at high temperatures.
ISSN:0916-8451
1347-6947
DOI:10.1271/bbb.80853