Alcohol dehydrogenase gene ADH3 activates glucose alcoholic fermentation in genetically engineered Dekkera bruxellensis yeast

Anna Judith Schifferdecker; Juozas Siurkus; Mikael Rørdam Andersen; Dorte Jørck-Ramberg; Zhihao Ling; Nerve Zhou; James E. Blevins; Andriy A. Sibirny; Jure Piškur; Olena P Ishchuk

doi:10.1007/s00253-015-7266-x

Alcohol dehydrogenase gene ADH3 activates glucose alcoholic fermentation in genetically engineered Dekkera bruxellensis yeast

Anna Judith Schifferdecker, Juozas Siurkus, Mikael Rørdam Andersen, Dorte Jørck-Ramberg, Zhihao Ling, Nerve Zhou, James E. Blevins, Andriy A. Sibirny, Jure Piškur, Olena P Ishchuk

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Abstract

Dekkera bruxellensis is a non-conventional Crabtree-positive yeast with a good ethanol production capability. Compared to Saccharomyces cerevisiae, its tolerance to acidic pH and its utilization of alternative carbon sources make it a promising organism for producing biofuel. In this study, we developed an auxotrophic transformation system and an expression vector, which enabled the manipulation of D. bruxellensis, thereby improving its fermentative performance. Its gene ADH3, coding for alcohol dehydrogenase, was cloned and overexpressed under the control of the strong and constitutive promoter TEF1. Our recombinant D. bruxellensis strain displayed 1.4 and 1.7 times faster specific glucose consumption rate during aerobic and anaerobic glucose fermentations, respectively; it yielded 1.2 times and 1.5 times more ethanol than did the parental strain under aerobic and anaerobic conditions, respectively. The overexpression of ADH3 in D. bruxellensis also reduced the inhibition of fermentation by anaerobiosis, the "Custer effect". Thus, the fermentative capacity of D. bruxellensis could be further improved by metabolic engineering.

Original language	English
Journal	Applied Microbiology and Biotechnology
Volume	100
Issue number	7
Pages (from-to)	3219-3231
Number of pages	13
ISSN	0175-7598
DOIs	https://doi.org/10.1007/s00253-015-7266-x
Publication status	Published - 2016

Bibliographical note

This article is published with open access at Springerlink.com

An erratum to this article can be found at http://dx.doi.org/10.1007/s00253-016-7349-3.

Keywords

ADH3
Auxotrophic mutants
Dekkera bruxellensis
Gene expression
Glucose fermentation
Promoters
TEF1

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Alcohol dehydrogenase gene ADH3 activates glucose alcoholic fermentation in genetically engineered Dekkera bruxellensis yeastFinal published version, 2.35 MB
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Schifferdecker, A. J., Siurkus, J., Andersen, M. R., Jørck-Ramberg, D., Ling, Z., Zhou, N., Blevins, J. E., Sibirny, A. A., Piškur, J., & Ishchuk, O. P. (2016). Alcohol dehydrogenase gene ADH3 activates glucose alcoholic fermentation in genetically engineered Dekkera bruxellensis yeast. Applied Microbiology and Biotechnology, 100(7), 3219-3231. https://doi.org/10.1007/s00253-015-7266-x

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title = "Alcohol dehydrogenase gene ADH3 activates glucose alcoholic fermentation in genetically engineered Dekkera bruxellensis yeast",

abstract = "Dekkera bruxellensis is a non-conventional Crabtree-positive yeast with a good ethanol production capability. Compared to Saccharomyces cerevisiae, its tolerance to acidic pH and its utilization of alternative carbon sources make it a promising organism for producing biofuel. In this study, we developed an auxotrophic transformation system and an expression vector, which enabled the manipulation of D. bruxellensis, thereby improving its fermentative performance. Its gene ADH3, coding for alcohol dehydrogenase, was cloned and overexpressed under the control of the strong and constitutive promoter TEF1. Our recombinant D. bruxellensis strain displayed 1.4 and 1.7 times faster specific glucose consumption rate during aerobic and anaerobic glucose fermentations, respectively; it yielded 1.2 times and 1.5 times more ethanol than did the parental strain under aerobic and anaerobic conditions, respectively. The overexpression of ADH3 in D. bruxellensis also reduced the inhibition of fermentation by anaerobiosis, the {"}Custer effect{"}. Thus, the fermentative capacity of D. bruxellensis could be further improved by metabolic engineering. ",

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note = "This article is published with open access at Springerlink.com An erratum to this article can be found at http://dx.doi.org/10.1007/s00253-016-7349-3.",

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Schifferdecker, AJ, Siurkus, J, Andersen, MR, Jørck-Ramberg, D, Ling, Z, Zhou, N, Blevins, JE, Sibirny, AA, Piškur, J & Ishchuk, OP 2016, 'Alcohol dehydrogenase gene ADH3 activates glucose alcoholic fermentation in genetically engineered Dekkera bruxellensis yeast', Applied Microbiology and Biotechnology, vol. 100, no. 7, pp. 3219-3231. https://doi.org/10.1007/s00253-015-7266-x

Alcohol dehydrogenase gene ADH3 activates glucose alcoholic fermentation in genetically engineered Dekkera bruxellensis yeast. / Schifferdecker, Anna Judith; Siurkus, Juozas; Andersen, Mikael Rørdam et al.
In: Applied Microbiology and Biotechnology, Vol. 100, No. 7, 2016, p. 3219-3231.

Research output: Contribution to journal › Journal article › Research › peer-review

TY - JOUR

T1 - Alcohol dehydrogenase gene ADH3 activates glucose alcoholic fermentation in genetically engineered Dekkera bruxellensis yeast

AU - Schifferdecker, Anna Judith

AU - Siurkus, Juozas

AU - Andersen, Mikael Rørdam

AU - Jørck-Ramberg, Dorte

AU - Ling, Zhihao

AU - Zhou, Nerve

AU - Blevins, James E.

AU - Sibirny, Andriy A.

AU - Piškur, Jure

AU - Ishchuk, Olena P

N1 - This article is published with open access at Springerlink.com An erratum to this article can be found at http://dx.doi.org/10.1007/s00253-016-7349-3.

PY - 2016

Y1 - 2016

N2 - Dekkera bruxellensis is a non-conventional Crabtree-positive yeast with a good ethanol production capability. Compared to Saccharomyces cerevisiae, its tolerance to acidic pH and its utilization of alternative carbon sources make it a promising organism for producing biofuel. In this study, we developed an auxotrophic transformation system and an expression vector, which enabled the manipulation of D. bruxellensis, thereby improving its fermentative performance. Its gene ADH3, coding for alcohol dehydrogenase, was cloned and overexpressed under the control of the strong and constitutive promoter TEF1. Our recombinant D. bruxellensis strain displayed 1.4 and 1.7 times faster specific glucose consumption rate during aerobic and anaerobic glucose fermentations, respectively; it yielded 1.2 times and 1.5 times more ethanol than did the parental strain under aerobic and anaerobic conditions, respectively. The overexpression of ADH3 in D. bruxellensis also reduced the inhibition of fermentation by anaerobiosis, the "Custer effect". Thus, the fermentative capacity of D. bruxellensis could be further improved by metabolic engineering.

AB - Dekkera bruxellensis is a non-conventional Crabtree-positive yeast with a good ethanol production capability. Compared to Saccharomyces cerevisiae, its tolerance to acidic pH and its utilization of alternative carbon sources make it a promising organism for producing biofuel. In this study, we developed an auxotrophic transformation system and an expression vector, which enabled the manipulation of D. bruxellensis, thereby improving its fermentative performance. Its gene ADH3, coding for alcohol dehydrogenase, was cloned and overexpressed under the control of the strong and constitutive promoter TEF1. Our recombinant D. bruxellensis strain displayed 1.4 and 1.7 times faster specific glucose consumption rate during aerobic and anaerobic glucose fermentations, respectively; it yielded 1.2 times and 1.5 times more ethanol than did the parental strain under aerobic and anaerobic conditions, respectively. The overexpression of ADH3 in D. bruxellensis also reduced the inhibition of fermentation by anaerobiosis, the "Custer effect". Thus, the fermentative capacity of D. bruxellensis could be further improved by metabolic engineering.

KW - ADH3

KW - Auxotrophic mutants

KW - Dekkera bruxellensis

KW - Gene expression

KW - Glucose fermentation

KW - Promoters

KW - TEF1

U2 - 10.1007/s00253-015-7266-x

DO - 10.1007/s00253-015-7266-x

M3 - Journal article

C2 - 26743658

SN - 0175-7598

VL - 100

SP - 3219

EP - 3231

JO - Applied Microbiology and Biotechnology

JF - Applied Microbiology and Biotechnology

IS - 7

ER -

Alcohol dehydrogenase gene ADH3 activates glucose alcoholic fermentation in genetically engineered Dekkera bruxellensis yeast

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