Overexpression of NADH-dependent fumarate reductase improves d-xylose fermentation in recombinant Saccharomyces cerevisiae

Laura Salusjärvi, Sanna Kaunisto, Sami Holmström, Maija-Leena Vehkomäki, Kari Koivuranta, Juha-Pekka Pitkänen, Laura Ruohonen (Corresponding Author)

Research output: Contribution to journalArticleScientificpeer-review

10 Citations (Scopus)

Abstract

Deviation from optimal levels and ratios of redox cofactors NAD(H) and NADP(H) is common when microbes are metabolically engineered. The resulting redox imbalance often reduces the rate of substrate utilization as well as biomass and product formation. An example is the metabolism of d-xylose by recombinant Saccharomyces cerevisiae strains expressing xylose reductase and xylitol dehydrogenase encoding genes from Scheffersomyces stipitis. This pathway requires both NADPH and NAD+. The effect of overexpressing the glycosomal NADH-dependent fumarate reductase (FRD) of Trypanosoma brucei in d-xylose-utilizing S. cerevisiae alone and together with an endogenous, cytosol directed NADH-kinase (POS5?17) was studied as one possible solution to overcome this imbalance. Expression of FRD and FRD + POS5?17 resulted in 60 and 23 % increase in ethanol yield, respectively, on d-xylose under anaerobic conditions. At the same time, xylitol yield decreased in the FRD strain suggesting an improvement in redox balance. We show that fumarate reductase of T. brucei can provide an important source of NAD+ in yeast under anaerobic conditions, and can be useful for metabolic engineering strategies where the redox cofactors need to be balanced. The effects of FRD and NADH-kinase on aerobic and anaerobic d-xylose and d-glucose metabolism are discussed.
Original languageEnglish
Pages (from-to)1383-1392
Number of pages9
JournalJournal of industrial microbiology and biotechnology
Volume40
Issue number12
DOIs
Publication statusPublished - 2013
MoE publication typeA1 Journal article-refereed

Fingerprint

Xylose
Succinate Dehydrogenase
NAD
Yeast
Fermentation
Oxidation-Reduction
Saccharomyces cerevisiae
Trypanosoma brucei brucei
NADP
D-Xylulose Reductase
Phosphotransferases
Metabolism
Metabolic Engineering
Xylitol
Aldehyde Reductase
Metabolic engineering
Gene encoding
Cytosol
Biomass
Ethanol

Keywords

  • Fumarate reductase
  • NADH-kinase
  • Saccharomyces cerevisiae
  • NADH
  • NADPH
  • d-Xylose
  • metabolic engineering
  • d-Xylose fermentation

Cite this

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title = "Overexpression of NADH-dependent fumarate reductase improves d-xylose fermentation in recombinant Saccharomyces cerevisiae",
abstract = "Deviation from optimal levels and ratios of redox cofactors NAD(H) and NADP(H) is common when microbes are metabolically engineered. The resulting redox imbalance often reduces the rate of substrate utilization as well as biomass and product formation. An example is the metabolism of d-xylose by recombinant Saccharomyces cerevisiae strains expressing xylose reductase and xylitol dehydrogenase encoding genes from Scheffersomyces stipitis. This pathway requires both NADPH and NAD+. The effect of overexpressing the glycosomal NADH-dependent fumarate reductase (FRD) of Trypanosoma brucei in d-xylose-utilizing S. cerevisiae alone and together with an endogenous, cytosol directed NADH-kinase (POS5?17) was studied as one possible solution to overcome this imbalance. Expression of FRD and FRD + POS5?17 resulted in 60 and 23 {\%} increase in ethanol yield, respectively, on d-xylose under anaerobic conditions. At the same time, xylitol yield decreased in the FRD strain suggesting an improvement in redox balance. We show that fumarate reductase of T. brucei can provide an important source of NAD+ in yeast under anaerobic conditions, and can be useful for metabolic engineering strategies where the redox cofactors need to be balanced. The effects of FRD and NADH-kinase on aerobic and anaerobic d-xylose and d-glucose metabolism are discussed.",
keywords = "Fumarate reductase, NADH-kinase, Saccharomyces cerevisiae, NADH, NADPH, d-Xylose, metabolic engineering, d-Xylose fermentation",
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Overexpression of NADH-dependent fumarate reductase improves d-xylose fermentation in recombinant Saccharomyces cerevisiae. / Salusjärvi, Laura; Kaunisto, Sanna; Holmström, Sami; Vehkomäki, Maija-Leena; Koivuranta, Kari; Pitkänen, Juha-Pekka; Ruohonen, Laura (Corresponding Author).

In: Journal of industrial microbiology and biotechnology, Vol. 40, No. 12, 2013, p. 1383-1392.

Research output: Contribution to journalArticleScientificpeer-review

TY - JOUR

T1 - Overexpression of NADH-dependent fumarate reductase improves d-xylose fermentation in recombinant Saccharomyces cerevisiae

AU - Salusjärvi, Laura

AU - Kaunisto, Sanna

AU - Holmström, Sami

AU - Vehkomäki, Maija-Leena

AU - Koivuranta, Kari

AU - Pitkänen, Juha-Pekka

AU - Ruohonen, Laura

PY - 2013

Y1 - 2013

N2 - Deviation from optimal levels and ratios of redox cofactors NAD(H) and NADP(H) is common when microbes are metabolically engineered. The resulting redox imbalance often reduces the rate of substrate utilization as well as biomass and product formation. An example is the metabolism of d-xylose by recombinant Saccharomyces cerevisiae strains expressing xylose reductase and xylitol dehydrogenase encoding genes from Scheffersomyces stipitis. This pathway requires both NADPH and NAD+. The effect of overexpressing the glycosomal NADH-dependent fumarate reductase (FRD) of Trypanosoma brucei in d-xylose-utilizing S. cerevisiae alone and together with an endogenous, cytosol directed NADH-kinase (POS5?17) was studied as one possible solution to overcome this imbalance. Expression of FRD and FRD + POS5?17 resulted in 60 and 23 % increase in ethanol yield, respectively, on d-xylose under anaerobic conditions. At the same time, xylitol yield decreased in the FRD strain suggesting an improvement in redox balance. We show that fumarate reductase of T. brucei can provide an important source of NAD+ in yeast under anaerobic conditions, and can be useful for metabolic engineering strategies where the redox cofactors need to be balanced. The effects of FRD and NADH-kinase on aerobic and anaerobic d-xylose and d-glucose metabolism are discussed.

AB - Deviation from optimal levels and ratios of redox cofactors NAD(H) and NADP(H) is common when microbes are metabolically engineered. The resulting redox imbalance often reduces the rate of substrate utilization as well as biomass and product formation. An example is the metabolism of d-xylose by recombinant Saccharomyces cerevisiae strains expressing xylose reductase and xylitol dehydrogenase encoding genes from Scheffersomyces stipitis. This pathway requires both NADPH and NAD+. The effect of overexpressing the glycosomal NADH-dependent fumarate reductase (FRD) of Trypanosoma brucei in d-xylose-utilizing S. cerevisiae alone and together with an endogenous, cytosol directed NADH-kinase (POS5?17) was studied as one possible solution to overcome this imbalance. Expression of FRD and FRD + POS5?17 resulted in 60 and 23 % increase in ethanol yield, respectively, on d-xylose under anaerobic conditions. At the same time, xylitol yield decreased in the FRD strain suggesting an improvement in redox balance. We show that fumarate reductase of T. brucei can provide an important source of NAD+ in yeast under anaerobic conditions, and can be useful for metabolic engineering strategies where the redox cofactors need to be balanced. The effects of FRD and NADH-kinase on aerobic and anaerobic d-xylose and d-glucose metabolism are discussed.

KW - Fumarate reductase

KW - NADH-kinase

KW - Saccharomyces cerevisiae

KW - NADH

KW - NADPH

KW - d-Xylose

KW - metabolic engineering

KW - d-Xylose fermentation

U2 - 10.1007/s10295-013-1344-9

DO - 10.1007/s10295-013-1344-9

M3 - Article

VL - 40

SP - 1383

EP - 1392

JO - Journal of industrial microbiology and biotechnology

JF - Journal of industrial microbiology and biotechnology

SN - 1367-5435

IS - 12

ER -