Construction of xylose-assimilating Saccharomyces cerevisiae

Tantirungkij, Manee; Nakashima, Noriyuki; Seki, Tatsuji; Yoshida, Takemi

doi:10.1016/0922-338x(93)90214-s

Cited by 154 publications

(82 citation statements)

References 26 publications

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“…For an economically feasible industrial process for ethanol production from plant biomass in lignocellulose hydrolysates, it is necessary to ferment all sugars present (von Sivers & Zacchi, 1995). Efforts to establish a xylose-utilizing pathway in S. cerevisiae by insertion of the genes encoding xylose reductase and xylitol dehydrogenase from Pichia stipitis or other organisms have resulted in only poor ethanol production from xylose (Ko$ tter & Ciriacy, 1993 ;Tantirungkij et al, 1993 ;Walfridsson et al, 1995). Various steps, including the uptake of xylose, have been suggested to limit the metabolism of xylose in metabolically engineered S. cerevisiae (Ko$ tter & Ciriacy, 1993 ;Eliasson et al, 2000).…”

Section: Introductionmentioning

confidence: 99%

Characterization of the xylose-transporting properties of yeast hexose transporters and their influence on xylose utilization

et al. 2002

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For an economically feasible production of ethanol from plant biomass by microbial cells, the fermentation of xylose is important. As xylose uptake might be a limiting step for xylose fermentation by recombinant xyloseutilizing Saccharomyces cerevisiae cells a study of xylose uptake was performed. After deletion of all of the 18 hexose-transporter genes, the ability of the cells to take up and to grow on xylose was lost. Reintroduction of individual hexose-transporter genes in this strain revealed that at intermediate xylose concentrations the yeast high-and intermediate-affinity transporters Hxt4, Hxt5, Hxt7 and Gal2 are important xylose-transporting proteins. Several heterologous monosaccharide transporters from bacteria and plant cells did not confer sufficient uptake activity to restore growth on xylose. Overexpression of the xylose-transporting proteins in a xylose-utilizing PUA yeast strain did not result in faster growth on xylose under aerobic conditions nor did it enhance the xylose fermentation rate under anaerobic conditions. The results of this study suggest that xylose uptake does not determine the xylose flux under the conditions and in the yeast strains investigated.

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Section: Introductionmentioning

confidence: 99%

Characterization of the xylose-transporting properties of yeast hexose transporters and their influence on xylose utilization

et al. 2002

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“…Recombinant xylose utilizing S. cerevisiae strains producing XR and XDH from Pichia stipitis have been constructed (Kötter and Ciriacy, 1993;Tantirungkij et al, 1993;Walfridsson et al, 1997). However, in these strains a major product has been xylitol, caused by a co-factor imbalance between the NAD(P)H consuming XR (Rizzi et al, 1988) and the NADH forming XDH (Rizzi et al, 1989) reactions (Bruinenberg et al, 1983;Kötter and Ciriacy, 1993).…”

Section: Introductionmentioning

confidence: 99%

Endogenous NADPH‐dependent aldose reductase activity influences product formation during xylose consumption in recombinant Saccharomyces cerevisiae

Bjerre

Jeppsson

Hahn‐Hägerdal

et al. 2003

Yeast

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Introduction of the xylose pathway from Pichia stipitis into Saccharomyces cerevisiae enables xylose utilization in recombinant S. cerevisiae. However, xylitol is a major by-product. An endogenous aldo-keto reductase, encoded by the GRE3 gene, was expressed at different levels in recombinant S. cerevisiae strains to investigate its effect on xylose utilization. In a recombinant S. cerevisiae strain producing only xylitol dehydrogenase (XDH) from P. stipitis and an extra copy of the endogenous xylulokinase (XK), ethanol formation from xylose was mediated by Gre3p, capable of reducing xylose to xylitol. When the GRE3 gene was overexpressed in this strain, the xylose consumption and ethanol formation increased by 29% and 116%, respectively. When the GRE3 gene was deleted in the recombinant xylose-fermenting S. cerevisiae strain TMB3001 (which possesses xylose reductase and XDH from P. stipitis, and an extra copy of endogenous XK), the xylitol yield decreased by 49% and the ethanol yield increased by 19% in anaerobic continuous culture with a glucose/xylose mixture. Biomass was reduced by 31% in strains where GRE3 was deleted, suggesting that fine-tuning of GRE3 expression is the preferred choice rather than deletion.

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“…1). However, this pathway introduces a problematic redox cofactor imbalance because XR can use both NADPH and nicotinamide adenine dinucleotide (NADH), whereas XDH uses NAD þ only, leading to high xylitol accumulation and low ethanol yields [17][18][19][20] . Xylitol formation could be reduced by providing sufficient oxygen 21 .…”

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confidence: 99%

Enhanced biofuel production through coupled acetic acid and xylose consumption by engineered yeast

et al. 2013

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The anticipation for substituting conventional fossil fuels with cellulosic biofuels is growing in the face of increasing demand for energy and rising concerns of greenhouse gas emissions. However, commercial production of cellulosic biofuel has been hampered by inefficient fermentation of xylose and the toxicity of acetic acid, which constitute substantial portions of cellulosic biomass. Here we use a redox balancing strategy to enable efficient xylose fermentation and simultaneous in situ detoxification of cellulosic feedstocks. By combining a nicotinamide adenine dinucleotide (NADH)-consuming acetate consumption pathway and an NADH-producing xylose utilization pathway, engineered yeast converts cellulosic sugars and toxic levels of acetate together into ethanol under anaerobic conditions. The results demonstrate a breakthrough in making efficient use of carbon compounds in cellulosic biomass and present an innovative strategy for metabolic engineering whereby an undesirable redox state can be exploited to drive desirable metabolic reactions, even improving productivity and yield.

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Construction of xylose-assimilating Saccharomyces cerevisiae

Cited by 154 publications

References 26 publications

Characterization of the xylose-transporting properties of yeast hexose transporters and their influence on xylose utilization

Characterization of the xylose-transporting properties of yeast hexose transporters and their influence on xylose utilization

Endogenous NADPH‐dependent aldose reductase activity influences product formation during xylose consumption in recombinant Saccharomyces cerevisiae

Enhanced biofuel production through coupled acetic acid and xylose consumption by engineered yeast

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