Xylose isomerase activity influences xylose fermentation with recombinant Saccharomyces cerevisiae strains expressing mutated xylA from Thermus thermophilus

Lönn, Anna; Bjerre, Karin; Otero, Ricardo R. Cordero; Zyl, Willem H. van; Hahn‐Hägerdal, Bärbel

doi:10.1016/s0141-0229(03)00024-3

Cited by 43 publications

(24 citation statements)

References 38 publications

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“…The integrating vector YIpXR/XDH/XK (Eliasson et al, 2000) was linearized with the restriction enzyme PstI to target the integration in the HIS3 locus in the chromosome of TMB3105 (Lönn et al, 2003), generating TMB3120 after selection for histidine prototrophy.…”

Section: Construction Of S Cerevisiae Tmb3120mentioning

confidence: 99%

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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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Section: Construction Of S Cerevisiae Tmb3120mentioning

confidence: 99%

“…The GRE3 gene was first deleted in the CEN.PK113-7A strain, generating TMB3105( gre3 ) (Lönn et al, 2003). No detectable XR activity and xylitol as the only product was observed (Tables 2, 3).…”

Section: Effect Of Gre3 Deletion and Overexpression In Recombinant Xymentioning

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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“…The first step is to introduce xylose-utilizing pathways such as xylose reductase/xylitol dehydrogenase (XR/XDH) or xylose isomerase (XI) from yeast or bacteria/fungi, respectively. Even though bacterial/fungal xylA genes, coding for XI, have been successfully expressed in S. cerevisiae (Brat et al, 2009;Ha et al, 2011;Karhumaa et al, 2005;Kuyper et al, 2003;Kuyper et al, 2005;Lönn et al, 2003;Walfridsson et al, 1996), it has been reported that the XR/XDH pathway can offer higher metabolic fluxes than the XI pathway (Karhumaa et al, 2007b). In terms of ethanol yield, however, the XR/XDH pathway has been considered inferior to the XI pathway because the dual enzyme pathway is accompanied by oxidation/ reduction of different cofactors and accumulates xylitol as an intermediate product (Akinterinwa and Cirino, 2009;Ho et al, 1998;Kotter and Ciriacy, 1993;Tantirungkij et al, 1993;Walfridsson et al, 1997).…”

Section: Introductionmentioning

confidence: 99%

High expression of XYL2 coding for xylitol dehydrogenase is necessary for efficient xylose fermentation by engineered Saccharomyces cerevisiae

Kim

Kong

et al. 2012

Metabolic Engineering

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“…By respiring NADH and supplying NADPH through the pentose phosphate pathway, which is rather active in the xylose-utilizing yeast P. stipitis (8), yeast metabolism can efficiently drive these coupled redox reactions for respirative growth but not for fermentative growth. Functional expression of xylose isomerase in recombinant xylose-utilizing S. cerevisiae, however, does not enable anaerobic growth on xylose (13,21,33,50), which indicates that there must be limitations beyond redox balancing.…”

mentioning

confidence: 99%

Molecular Basis for Anaerobic Growth of Saccharomyces cerevisiae on Xylose, Investigated by Global Gene Expression and Metabolic Flux Analysis

Sonderegger

Jeppsson

Hahn‐Hägerdal

et al. 2004

Appl Environ Microbiol

120

109

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Yeast xylose metabolism is generally considered to be restricted to respirative conditions because the two-step oxidoreductase reactions from xylose to xylulose impose an anaerobic redox imbalance. We have recently developed, however, a Saccharomyces cerevisiae strain that is at present the only known yeast capable of anaerobic growth on xylose alone. Using transcriptome analysis of aerobic chemostat cultures grown on xylose-glucose mixtures and xylose alone, as well as a combination of global gene expression and metabolic flux analysis of anaerobic chemostat cultures grown on xylose-glucose mixtures, we identified the distinguishing characteristics of this unique phenotype. First, the transcript levels and metabolic fluxes throughout central carbon metabolism were significantly higher than those in the parent strain, and they were most pronounced in the xylose-specific, pentose phosphate, and glycerol pathways. Second, differential expression of many genes involved in redox metabolism indicates that increased cytosolic NADPH formation and NADH consumption enable a higher flux through the two-step oxidoreductase reaction of xylose to xylulose in the mutant. Redox balancing is apparently still a problem in this strain, since anaerobic growth on xylose could be improved further by providing acetoin as an external NADH sink. This improved growth was accompanied by an increased ATP production rate and was not accompanied by higher rates of xylose uptake or cytosolic NADPH production. We concluded that anaerobic growth of the yeast on xylose is ultimately limited by the rate of ATP production and not by the redox balance per se, although the redox imbalance, in turn, limits ATP production.

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Xylose isomerase activity influences xylose fermentation with recombinant Saccharomyces cerevisiae strains expressing mutated xylA from Thermus thermophilus

Cited by 43 publications

References 38 publications

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

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

High expression of XYL2 coding for xylitol dehydrogenase is necessary for efficient xylose fermentation by engineered Saccharomyces cerevisiae

Molecular Basis for Anaerobic Growth of Saccharomyces cerevisiae on Xylose, Investigated by Global Gene Expression and Metabolic Flux Analysis

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