Stereochemistry of Furfural Reduction by a
            <i>Saccharomyces cerevisiae</i>
            Aldehyde Reductase That Contributes to
            <i>In Situ</i>
            Furfural Detoxification

Bowman, Michael J.; Jordan, Douglas B.; Vermillion, Karl E.; Braker, Jay D.; Moon, Jon; Liu, Z. Lewis

doi:10.1128/aem.00542-10

Cited by 29 publications

(14 citation statements)

References 40 publications

(37 reference statements)

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“…In addition, OYE3 is a conserved NADPH oxidoreductase and is involved in metabolism and oxidative stress response [42]. For the fructose growth condition, ARI1 - an NADPH-dependent aldehyde reductase - is involved in regulation of fermentation [43]. Similar to glucose, the relative concentration of hexose transporter HXT3 [37] was also considered greatly altered in relative abundance in the fructose culture (Supplementary Figure 3D).…”

Section: Resultsmentioning

confidence: 99%

Quantitative mass spectrometry-based multiplexing compares the abundance of 5000 S. cerevisiae proteins across 10 carbon sources

Paulo

O’Connell

Everley

et al. 2016

Journal of Proteomics

188

197

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The budding yeast Saccharomyces cerevisiae is a model system for investigating biological processes. Cellular processes are known to be dysregulated because of shifts in carbon sources. However, the comprehensive proteomic alterations thereof have not been fully investigated. Here we examined proteomic alterations in S. cerevisiae due to the adaptation of yeast from glucose to nine different carbon sources – maltose, trehalose, fructose, sucrose, glycerol, acetate, pyruvate, lactic acid, and oleate. Isobaric tag-based mass spectrometry techniques are at the forefront of global proteomic investigations. As such, we used a TMT10-plex strategy to study multiple growth conditions in a single experiment. The SPS-MS3 method on an Orbitrap Fusion Lumos mass spectrometer enabled the quantification of over 5000 yeast proteins across ten carbon sources at a 1% protein-level FDR. On average, the proteomes of yeast cultured in fructose and sucrose deviated the least from those cultured in glucose. As expected, gene ontology classification revealed the major alteration in protein abundances occurred in metabolic pathways and mitochondrial proteins. Our protocol lays the groundwork for further investigation of carbon source-induced protein alterations. Additionally, these data offer a hypothesis-generating resource for future studies aiming to investigate both characterized and uncharacterized genes.

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Section: Resultsmentioning

confidence: 99%

Quantitative mass spectrometry-based multiplexing compares the abundance of 5000 S. cerevisiae proteins across 10 carbon sources

Paulo

O’Connell

Everley

et al. 2016

Journal of Proteomics

188

197

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“…NADPH generated via the oxidative branch of the PPP has been shown to be required for growth in the presence of either furfural or HMF (Gorsich et al , 2006). Additionally, NADPH‐dependent furfural reduction has been observed in S. cerevisiae (Bowman et al , 2010; Liu and Moon, 2009). We hypothesized that when cells are provided xylose as the only carbon source, sensitivity to these compounds will be increased.…”

Section: Resultsmentioning

confidence: 99%

Saccharomyces cerevisiae engineered for xylose metabolism requires gluconeogenesis and the oxidative branch of the pentose phosphate pathway for aerobic xylose assimilation

Hector¹,

Mertens²,

Bowman³

et al. 2011

Yeast

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Saccharomyces strains engineered to ferment xylose using Scheffersomyces stipitis xylose reductase (XR) and xylitol dehydrogenase (XDH) genes appear to be limited by metabolic imbalances, due to differing cofactor specificities of XR and XDH. The S. stipitis XR, which uses both NADH and NADPH, is hypothesized to reduce the cofactor imbalance, allowing xylose fermentation in this yeast. However, unadapted S. cerevisiae strains expressing this XR grow poorly on xylose, suggesting that metabolism is still imbalanced, even under aerobic conditions. In this study, we investigated the possible reasons for this imbalance by deleting genes required for NADPH production and gluconeogenesis in S. cerevisiae. S. cerevisiae cells expressing the XR-XDH, but not a xylose isomerase, pathway required the oxidative branch of the pentose phosphate pathway (PPP) and gluconeogenic production of glucose-6-P for xylose assimilation. The requirement for generating glucose-6-P from xylose was also shown for Kluyveromyces lactis. When grown in xylose medium, both K. lactis and S. stipitis showed increases in enzyme activity required for producing glucose-6-P. Thus, natural xylose-assimilating yeast respond to xylose, in part, by upregulating enzymes required for recycling xylose back to glucose-6-P for the production of NADPH via the oxidative branch of the PPP. Finally, we show that induction of these enzymes correlated with increased tolerance to the NADPH-depleting compound diamide and the fermentation inhibitors furfural and hydroxymethyl furfural; S. cerevisiae was not able to increase enzyme activity for glucose-6-P production when grown in xylose medium and was more sensitive to these inhibitors in xylose medium compared to glucose. Published in

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“…Furfural is generally considered the most potent inhibitor to various microbial catalysts such as E. coli, Z. mobilis , and yeast (Zaldivar et al, 1999; Liu et al, 2004, 2005, 2008; Gorsich et al, 2006; Allen et al, 2010; Bowman et al, 2010; He et al, 2012; Huang et al, 2012; Franden et al, 2013). The maximum amount of furfural was 2 g/L in the slurries in this study.…”

Section: Resultsmentioning

confidence: 99%

Connecting lignin-degradation pathway with pre-treatment inhibitor sensitivity of Cupriavidus necator

Wang¹,

Yang²,

Hunsinger³

et al. 2014

Front. Microbiol.

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To produce lignocellulosic biofuels economically, the complete release of monomers from the plant cell wall components, cellulose, hemicellulose, and lignin, through pre-treatment and hydrolysis (both enzymatic and chemical), and the efficient utilization of these monomers as carbon sources, is crucial. In addition, the identification and development of robust microbial biofuel production strains that can tolerate the toxic compounds generated during pre-treatment and hydrolysis is also essential. In this work, Cupriavidus necator was selected due to its capabilities for utilizing lignin monomers and producing polyhydroxylbutyrate (PHB), a bioplastic as well as an advanced biofuel intermediate. We characterized the growth kinetics of C. necator in pre-treated corn stover slurry as well as individually in the pre-sence of 11 potentially toxic compounds in the saccharified slurry. We found that C. necator was sensitive to the saccharified slurry produced from dilute acid pre-treated corn stover. Five out of 11 compounds within the slurry were characterized as toxic to C. necator, namely ammonium acetate, furfural, hydroxymethylfurfural (HMF), benzoic acid, and p-coumaric acid. Aldehydes (e.g., furfural and HMF) were more toxic than the acetate and the lignin degradation products benzoic acid and p-coumaric acid; furfural was identified as the most toxic compound. Although toxic to C. necator at high concentration, ammonium acetate, benzoic acid, and p-coumaric acid could be utilized by C. necator with a stimulating effect on C. necator growth. Consequently, the lignin degradation pathway of C. necator was reconstructed based on genomic information and literature. The efficient conversion of intermediate catechol to downstream products of cis,cis-muconate or 2-hydroxymuconate-6-semialdehyde may help improve the robustness of C. necator to benzoic acid and p-coumaric acid as well as improve PHB productivity.

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Stereochemistry of Furfural Reduction by a Saccharomyces cerevisiae Aldehyde Reductase That Contributes to In Situ Furfural Detoxification

Abstract: Ari1p from

Cited by 29 publications

References 40 publications

Quantitative mass spectrometry-based multiplexing compares the abundance of 5000 S. cerevisiae proteins across 10 carbon sources

Quantitative mass spectrometry-based multiplexing compares the abundance of 5000 S. cerevisiae proteins across 10 carbon sources

Saccharomyces cerevisiae engineered for xylose metabolism requires gluconeogenesis and the oxidative branch of the pentose phosphate pathway for aerobic xylose assimilation

Connecting lignin-degradation pathway with pre-treatment inhibitor sensitivity of Cupriavidus necator

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