The expression in Saccharomyces cerevisiae of a glucose/xylose symporter from Candida intermedia is affected by the presence of a glucose/xylose facilitator

Leandro, Maria; Spencer‐Martins, I.; Gonçalves, Paula

doi:10.1099/mic.0.2007/015511-0

Cited by 36 publications

(20 citation statements)

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“…Of the heterologous transport proteins, the two C. intermedia transporters are probably the most extensively studied (28,29,34). Previous reports focus on only the glucose and xylose transport capabilities of these proteins.…”

Section: Vol 77 2011 Survey For Heterologous Sugar Transport Proteimentioning

confidence: 99%

Functional Survey for Heterologous Sugar Transport Proteins, Using Saccharomyces cerevisiae as a Host

Young

Poucher

Comer

et al. 2011

Appl Environ Microbiol

146

136

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Molecular transport is a key process in cellular metabolism. This step is often limiting when using a nonnative carbon source, as exemplified by xylose catabolism in Saccharomyces cerevisiae. As a step toward addressing this limitation, this study seeks to characterize monosaccharide transport preference and efficiency. A group of 26 known and putative monosaccharide transport proteins was expressed in a recombinant Saccharomyces cerevisiae host unable to transport several monosaccharides. A growth-based assay was used to detect transport capacity across six different carbon sources (glucose, xylose, galactose, fructose, mannose, and ribose). A mixed glucose-and-xylose cofermentation was performed to determine substrate preference. These experiments identified 10 transporter proteins that function as transporters of one or more of these sugars. Most of these proteins exhibited broad substrate ranges, and glucose was preferred in all cases. The broadest transporters confer the highest growth rates and strongly prefer glucose. This study reports the first molecular characterization of the annotated XUT genes of Scheffersomyces stipitis and open reading frames from the yeasts Yarrowia lipolytica and Debaryomyces hansenii. Finally, a phylogenetic analysis demonstrates that transporter function clusters into three distinct groups. One particular group comprised of D. hansenii XylHP and S. stipitis XUT1 and XUT3 demonstrated moderate transport efficiency and higher xylose preferences.

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Section: Vol 77 2011 Survey For Heterologous Sugar Transport Proteimentioning

confidence: 99%

Functional Survey for Heterologous Sugar Transport Proteins, Using Saccharomyces cerevisiae as a Host

Young

Poucher

Comer

et al. 2011

Appl Environ Microbiol

146

136

View full text Add to dashboard Cite

show abstract

“…Moreover, most of the known pentose transporters were shown to be able to differentiate between D-xylose and L-arabinose. For example, Gxf1 and Gxs1 can transport D-xylose but not L-arabinose, while the D-xylose/L-arabinose transporter (XAT-1) transports Larabinose much better than D-xylose (16,20,24). To date, only a few L-arabinose-specific transporters have been reported, including two transporters for L-arabinose cloned from Ambrosiozyma monospora in 2011 (21).…”

mentioning

confidence: 99%

“…Recently, multiple novel transporters from pentose-assimilating microorganisms have been discovered (19)(20)(21)(22)(23). In addition, it was shown that pentose utilization could be improved by increasing its uptake in S. cerevisiae via heterologous expression of glucose/xylose facilitator 1 (Gxf1) and glucose/xylose symporter 1 (Gxs1) from Candida intermedia (24) or by overexpression of the endogenous Gal2 (25). Alternative approaches using transporter engineering and directed evolution found that their substrate specificity could be rewired, and Dglucose inhibition-free D-xylose transporters were engineered through modification of protein motifs (26,27).…”

mentioning

confidence: 99%

Functional Analysis of Two l -Arabinose Transporters from Filamentous Fungi Reveals Promising Characteristics for Improved Pentose Utilization in Saccharomyces cerevisiae

Cai

et al. 2015

Appl Environ Microbiol

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cLimited uptake is one of the bottlenecks for L-arabinose fermentation from lignocellulosic hydrolysates in engineered Saccharomyces cerevisiae. This study characterized two novel L-arabinose transporters, LAT-1 from Neurospora crassa and MtLAT-1 from Myceliophthora thermophila. Although the two proteins share high identity (about 83%), they display different substrate specificities. Sugar transport assays using the S. cerevisiae strain EBY.VW4000 indicated that LAT-1 accepts a broad substrate spectrum. In contrast, MtLAT-1 appeared much more specific for L-arabinose. Determination of the kinetic properties of both transporters revealed that the K m values of LAT-1 and MtLAT-1 for L-arabinose were 58.12 ؎ 4.06 mM and 29.39 ؎ 3.60 mM, respectively, with corresponding V max values of 116.7 ؎ 3.0 mmol/h/g dry cell weight (DCW) and 10.29 ؎ 0.35 mmol/h/g DCW, respectively. In addition, both transporters were found to use a proton-coupled symport mechanism and showed only partial inhibition by D-glucose during L-arabinose uptake. Moreover, LAT-1 and MtLAT-1 were expressed in the S. cerevisiae strain BSW2AP containing an L-arabinose metabolic pathway. Both recombinant strains exhibited much faster L-arabinose utilization, greater biomass accumulation, and higher ethanol production than the control strain. In conclusion, because of higher maximum velocities and reduced inhibition by D-glucose, the genes for the two characterized transporters are promising targets for improved L-arabinose utilization and fermentation in S. cerevisiae. Biorefining of lignocellulosic biomass has attracted considerable attention in recent years because of its abundance, sustainability and potential environmental benefits (1, 2). The main sugars in hydrolysates from currently used feedstocks are a mixture of D-glucose, D-xylose, and L-arabinose (3). For a cost-effective conversion of biomass into fuels or chemicals, the fermentative organisms are required to utilize all three sugars efficiently. Even though many organisms are able to natively convert these substrates, the most commonly selected microbe is Saccharomyces cerevisiae (baker's yeast) because of its high ethanol productivity and high tolerance for inhibitors present in biomass hydrolysates, as well as the fact that it is amenable to genetic engineering (4-6).Wild-type S. cerevisiae cannot utilize the two pentose sugars D-xylose and L-arabinose. Thus, improving import and the intracellular pentose utilization efficiency is very critical, and intensive efforts have been made to do so by yeast metabolic engineering (7-13). Sugar uptake is the initial step for its utilization, and therefore, efficient molecular transport is a prerequisite to achieve enhanced fermentation rates in the presence of a working pentose metabolism pathway. Baker's yeast is able to absorb pentoses through its native hexose transporters, such as Hxt5 and Hxt7, and the galactose transporter Gal2 (14-16). However, these transporters show higher affinity for hexoses, and thus, it was suggested that D-glucose impa...

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“…GXF1 and GXS1 are transporters that play an important role in enhancing the utilization of glucose 10-fold higher than xylose. The expression of Gxs1and Gxf1 is directly proportional to the concentration of glucose [112]. At the high concentrations of xylose, Gxf1 had no distinguishable influence on a xylose engineered S. cerevisiae strain, but it boosted growth remarkably at low concentrations [148].…”

Section: Xylose Transportmentioning

confidence: 99%

“…Likewise, the overexpression of P. stipitis and other fungal sugar transporters can enhance the performance of engineered S. cerevisiae to grow and uptake xylose [109][110][111][112]. However, additional regulatory steps are required to engineer an efficient ethanol producer S. cerevisiae strain, because the mechanism for production of ethanol in response to xylose in S. cerevisiae is missing [113].…”

Section: New Yeast For Lignocelluloses Bioconversionmentioning

confidence: 99%

Bioethanol a Microbial Biofuel Metabolite; New Insights of Yeasts Metabolic Engineering

et al. 2018

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Scarcity of the non-renewable energy sources, global warming, environmental pollution, and raising the cost of petroleum are the motive for the development of renewable, eco-friendly fuels production with low costs. Bioethanol production is one of the promising materials that can subrogate the petroleum oil, and it is considered recently as a clean liquid fuel or a neutral carbon. Diverse microorganisms such as yeasts and bacteria are able to produce bioethanol on a large scale, which can satisfy our daily needs with cheap and applicable methods. Saccharomyces cerevisiae and Pichia stipitis are two of the pioneer yeasts in ethanol production due to their abilities to produce a high amount of ethanol. The recent focus is directed towards lignocellulosic biomass that contains 30-50% cellulose and 20-40% hemicellulose, and can be transformed into glucose and fundamentally xylose after enzymatic hydrolysis. For this purpose, a number of various approaches have been used to engineer different pathways for improving the bioethanol production with simultaneous fermentation of pentose and hexoses sugars in the yeasts. These approaches include metabolic and flux analysis, modeling and expression analysis, followed by targeted deletions or the overexpression of key genes. In this review, we highlight and discuss the current status of yeasts genetic engineering for enhancing bioethanol production, and the conditions that influence bioethanol production.

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The expression in Saccharomyces cerevisiae of a glucose/xylose symporter from Candida intermedia is affected by the presence of a glucose/xylose facilitator

Cited by 36 publications

References 50 publications

Functional Survey for Heterologous Sugar Transport Proteins, Using Saccharomyces cerevisiae as a Host

Functional Survey for Heterologous Sugar Transport Proteins, Using Saccharomyces cerevisiae as a Host

Functional Analysis of Two l -Arabinose Transporters from Filamentous Fungi Reveals Promising Characteristics for Improved Pentose Utilization in Saccharomyces cerevisiae

Bioethanol a Microbial Biofuel Metabolite; New Insights of Yeasts Metabolic Engineering

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