Control of xylose consumption by xylose transport in recombinant <i>Saccharomyces cerevisiae</i>

Gárdonyi, Márk; Jeppsson, Marie; Lidén, Gunnar; Gorwa-Grauslund, Marie-Françoise; Hahn‐Hägerdal, Bärbel

doi:10.1002/bit.10631

Cited by 79 publications

(65 citation statements)

References 33 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…To investigate whether the improved fermentation characteristics were indeed due to changes in sugar transport, zero trans-influx assays were performed using both the strain that was only metabolically engineered and the subsequently evolved strain [44]. The d-xylose uptake kinetics obtained for the metabolically engineered strain (K m 132 mM, V max 15.8 mmol (g dry weight) -1 h -1 ) were in agreement with other studies [22,39]. Strikingly, the d-xylose uptake kinetics of the evolved strain had changed drastically, with a 25% reduction in the K m (to 99 mM) and a twofold increase of V max to 32 mmol (g dry weight) -1 h -1 .…”

Section: Evolutionary Engineering Of D-xylose-consuming S Cerevisiaesupporting

confidence: 80%

Development of Efficient Xylose Fermentation in Saccharomyces cerevisiae: Xylose Isomerase as a Key Component

Maris

Winkler

Kuyper

et al. 2007

Advances in Biochemical Engineering/Biotechnology

168

160

View full text Add to dashboard Cite

Section: Evolutionary Engineering Of D-xylose-consuming S Cerevisiaesupporting

confidence: 80%

Development of Efficient Xylose Fermentation in Saccharomyces cerevisiae: Xylose Isomerase as a Key Component

Maris

Winkler

Kuyper

et al. 2007

Advances in Biochemical Engineering/Biotechnology

168

160

View full text Add to dashboard Cite

“…Transport is a bottleneck in xylose fermentation, at least at low concentrations, and certainly is exacerbated with improvements of intracellular xylose metabolism (15,16,21). Some heterologous xylose transporters have been identified (11,(31)(32)(33), and increased transporter expression has proven to be beneficial for xylose-fermentation performance (17,19,20).…”

Section: Discussionmentioning

confidence: 99%

“…This intrinsic transport is enough to enable growth on xylose and has not limited xylose consumption in early experiments (12). However, with improvements of intracellular xylose conversion being made (e.g., by overexpression of xylulokinase XKS1 and the PPP enzymes) (13,14), transport becomes a bottleneck, especially at low xylose concentrations (15)(16)(17)(18). In several studies, enhanced transport could be used to improve engineered strains further or could be seen as a result of evolutionary engineering (19)(20)(21)(22).…”

mentioning

confidence: 99%

Engineering of yeast hexose transporters to transport d -xylose without inhibition by d -glucose

Farwick

Bruder

Schadeweg

et al. 2014

Proc. Natl. Acad. Sci. U.S.A.

268

347

View full text Add to dashboard Cite

All known D-xylose transporters are competitively inhibited by D-glucose, which is one of the major reasons hampering simultaneous fermentation of D-glucose and D-xylose, two primary sugars present in lignocellulosic biomass. We have set up a yeast growthbased screening system for mutant D-xylose transporters that are insensitive to the presence of D-glucose. All of the identified variants had a mutation at either a conserved asparagine residue in transmembrane helix 8 or a threonine residue in transmembrane helix 5. According to a homology model of the yeast hexose transporter Gal2 deduced from the crystal structure of the D-xylose transporter XylE from Escherichia coli, both residues are found in the same region of the protein and are positioned slightly to the extracellular side of the central sugar-binding pocket. Therefore, it is likely that alterations sterically prevent D-glucose but not D-xylose from entering the pocket. In contrast, changing amino acids that are supposed to directly interact with the C6 hydroxymethyl group of D-glucose negatively affected transport of both D-glucose and D-xylose. Determination of kinetic properties of the mutant transporters revealed that Gal2-N376F had the highest affinity for D-xylose, along with a moderate transport velocity, and had completely lost the ability to transport hexoses. These transporter versions should prove valuable for glucose-xylose cofermentation in lignocellulosic hydrolysates by Saccharomyces cerevisiae and other biotechnologically relevant organisms. Moreover, our data contribute to the mechanistic understanding of sugar transport because the decisive role of the conserved asparagine residue for determining sugar specificity has not been recognized before.xylose transport | major facilitator superfamily | transporter engineering | pentose metabolism | HXT

show abstract

“…The Hxt4p, Hxt5p, Hxt7p and Gal2p have been shown to transport xylose, but overexpression of the individual transporter-encoding genes did not enhance the specific growth rate on xylose in recombinant S. cerevisiae (Hamacher et al, 2002). It has also been shown that transport only limits xylose consumption rate at low xylose concentrations (Gardonyi et al, 2003).…”

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

View full text Add to dashboard Cite

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.

show abstract

Control of xylose consumption by xylose transport in recombinant Saccharomyces cerevisiae

Abstract: Abstract:Saccharomyces cerevisiae TMB3001 has previ-

Cited by 79 publications

References 33 publications

Development of Efficient Xylose Fermentation in Saccharomyces cerevisiae: Xylose Isomerase as a Key Component

Development of Efficient Xylose Fermentation in Saccharomyces cerevisiae: Xylose Isomerase as a Key Component

Engineering of yeast hexose transporters to transport d -xylose without inhibition by d -glucose

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

Contact Info

Product

Resources

About