Central carbon metabolism of<i>Saccharomyces cerevisiae</i>in anaerobic, oxygen-limited and fully aerobic steady-state conditions and following a shift to anaerobic conditions

Wiebe, Marilyn G.; Rintala, Eija; Tamminen, Anu; Simolin, Helena; Salusjärvi, Laura; Toivari, Mervi; Kokkonen, Juha; Kiuru, Jari; Ketola, Raimo A.; Jouhten, Paula; Huuskonen, Anne; Maaheimo, Hannu; Ruohonen, Laura; Penttilä, Merja

doi:10.1111/j.1567-1364.2007.00234.x

Cited by 75 publications

(130 citation statements)

References 42 publications

(91 reference statements)

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“…1 and 2). Recent studies report on implications in the central carbon metabolism of Saccharomyces cerevisiae induced by aerobic and anaerobic conditions, and address the complexity of the role of oxygen in controlling cellular physiology in yeasts 29,33 .…”

Section: Discussionmentioning

confidence: 99%

Glucose and Fructose Fermentation by Wine Yeasts in Media Containing Structurally Complex Nitrogen Sources

Júnior¹,

Batistote²,

Ernandes

2008

Journal of the Institute of Brewing

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Glucose and fructose fermentations by industrial yeasts strains are strongly affected by both the structural complexity of the nitrogen source and the availability of oxygen. In this study two Saccharomyces cerevisiae industrial wine strains were grown, under shaken and static conditions, in a media containing either a) 20% (w/v) glucose, or b) 10% (w/v) fructose and 10% (w/v) glucose or c) 20% (w/v) fructose, all supplemented with nitrogen sources varying from a single ammonium salt (ammonium sulfate) to free amino acids (casamino acids) and peptides (peptone). Data suggest that a complex structured nitrogen source is not submitted to the same control mechanisms as those involved in the utilization of simpler structured nitrogen sources, and mutual interaction between carbon and nitrogen sources, including the mechanisms involved in the regulation of aerobic/anaerobic metabolism, may play an important role in defining yeast fermentation performance and the differing response to the structural complexity of the nitrogen source, with a strong impact on fermentation performance.

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Section: Discussionmentioning

confidence: 99%

Glucose and Fructose Fermentation by Wine Yeasts in Media Containing Structurally Complex Nitrogen Sources

Júnior¹,

Batistote²,

Ernandes

2008

Journal of the Institute of Brewing

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“…A cell density of 0.42 g/ml was used to convert mol/g CDW into mmol/liter (11). Lower and upper bounds of concentrations of pyruvate were set to 0.6 and 2.0 mM, respectively (62,70).…”

Section: Methodsmentioning

confidence: 99%

Limitations in Xylose-FermentingSaccharomyces cerevisiae, Made Evident through Comprehensive Metabolite Profiling and Thermodynamic Analysis

Klimacek

Krahulec

Sauer

et al. 2010

Appl Environ Microbiol

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Little is known about how the general lack of efficiency with which recombinant Saccharomyces cerevisiae strains utilize xylose affects the yeast metabolome. Quantitative metabolomics was therefore performed for two xylose-fermenting S. cerevisiae strains, BP000 and BP10001, both engineered to produce xylose reductase (XR), NAD ؉ -dependent xylitol dehydrogenase and xylulose kinase, and the corresponding wild-type strain CEN.PK 113-7D, which is not able to metabolize xylose. Contrary to BP000 expressing an NADPH-preferring XR, BP10001 expresses an NADH-preferring XR. An updated protocol of liquid chromatography/tandem mass spectrometry that was validated by applying internal 13 C-labeled metabolite standards was used to quantitatively determine intracellular pools of metabolites from the central carbon, energy, and redox metabolism and of eight amino acids. Metabolomic responses to different substrates, glucose (growth) or xylose (no growth), were analyzed for each strain. In BP000 and BP10001, flux through glycolysis was similarly reduced (ϳ27-fold) when xylose instead of glucose was metabolized. As a consequence, (i) most glycolytic metabolites were dramatically (<120-fold) diluted and (ii) energy and anabolic reduction charges were affected due to decreased ATP/AMP ratios (3-to 4-fold) and reduced NADP ؉ levels (ϳ3-fold), respectively. Contrary to that in BP000, the catabolic reduction charge was not altered in BP10001. This was due mainly to different utilization of NADH by XRs in BP000 (44%) and BP10001 (97%). Thermodynamic analysis complemented by enzyme kinetic considerations suggested that activities of pentose phosphate pathway enzymes and the pool of fructose-6-phosphate are potential factors limiting xylose utilization. Coenzyme and ATP pools did not rate limit flux through xylose pathway enzymes.Most of the biomass-to-ethanol processes that are currently advancing to the commercial-production scale rely on microbial strains for efficient fermentation of all sugars, that is, hexoses (mainly D-glucose) and pentoses (D-xylose, L-arabinose), contained in hydrolysates of lignocellulosic feedstocks (15, 36,65). The yeast Saccharomyces cerevisiae is among the top candidate microorganisms to be used for mixed hexosepentose fermentation (15). However, because neither xylose nor L-arabinose is a substrate naturally utilized by S. cerevisiae, extensive metabolic redesign was required to obtain strains producing pentose-derived ethanol in a useful yield (16,21, 37). Despite notable achievements, even the current best-performing yeast strains fall short in specific ethanol productivity (q ethanol ; 0.13 to 0.45 g ethanol/g biomass/h) from xylose compared to the corresponding q ethanol from glucose (1.2 g/g biomass/h) (33, 50). The low q ethanol for pentose fermentation is the result of at least two limiting factors. First of all, the yield coefficient (Y ethanol/sugar ) for consumption of pentose substrates is usually far lower (0.30 to 0.46) than theoretically expected (Y ethanol/xylose ϭ 0.51), reflecting the ...

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“…In addition to ethanol and carbon dioxide, the metabolism of glucose yields numerous products, including acetate, glycerol, carbohydrates, and macromolecules, such as RNA, DNA, and protein. Multiple independent lines of evidence indicate that the genetic background and the culture environment influence carbon metabolic flux and that mechanisms involving transcriptional regulation, such as glucose sensing, glucose repression, and oxygen responses, mediate the metabolic changes (14)(15)(16)(17)(18). Therefore, Rim15p may negatively regulate ethanol production through the activities of the downstream transcriptional activators Msn2/4p and Hsf1p, which induce the expression of genes involved in aerobic respiration, trehalose and glycogen metabolism, cell wall biosynthesis, pentose phosphate shuttling, and fatty acid metabolism (19,20).…”

mentioning

confidence: 99%

Inhibitory Role of Greatwall-Like Protein Kinase Rim15p in Alcoholic Fermentation via Upregulating the UDP-Glucose Synthesis Pathway in Saccharomyces cerevisiae

Watanabe

Zhou

Hirata

et al. 2016

Appl Environ Microbiol

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The high fermentation rate of Saccharomyces cerevisiae sake yeast strains is attributable to a loss-of-function mutation in the RIM15 gene, which encodes a Greatwall-family protein kinase that is conserved among eukaryotes. In the present study, we performed intracellular metabolic profiling analysis and revealed that deletion of the RIM15 gene in a laboratory strain impaired glucose-anabolic pathways through the synthesis of UDP-glucose (UDPG). Although Rim15p is required for the synthesis of trehalose and glycogen from UDPG upon entry of cells into the quiescent state, we found that Rim15p is also essential for the accumulation of cell wall ␤-glucans, which are also anabolic products of UDPG. Furthermore, the impairment of UDPG or 1,3-␤-glucan synthesis contributed to an increase in the fermentation rate. Transcriptional induction of PGM2 (phosphoglucomutase) and UGP1 (UDPG pyrophosphorylase) was impaired in Rim15p-deficient cells in the early stage of fermentation. These findings demonstrate that the decreased anabolism of glucose into UDPG and 1,3-␤-glucan triggered by a defect in the Rim15p-mediated upregulation of PGM2 and UGP1 redirects the glucose flux into glycolysis. Consistent with this, sake yeast strains with defective Rim15p exhibited impaired expression of PGM2 and UGP1 and decreased levels of ␤-glucans, trehalose, and glycogen during sake fermentation. We also identified a sake yeast-specific mutation in the glycogen synthesis-associated glycogenin gene GLG2, supporting the conclusion that the glucose-anabolic pathway is impaired in sake yeast. These findings demonstrate that downregulation of the UDPG synthesis pathway is a key mechanism accelerating alcoholic fermentation in industrially utilized S. cerevisiae sake strains. S ake yeast strains, which belong to the species Saccharomyces cerevisiae, are capable of achieving ethanol yields as high as 22 vol% in fermenting sake mash (1-3). This characteristic phenotype is attributed in part to their high and sustained maximum fermentation rates, as observed in batch cultures containing high concentrations of glucose (4), and is due to the continuous supply of fermentable sugars to yeast cells in sake mash via the degradation of rice starch by enzymes produced by Aspergillus oryzae. In recent studies of the representative sake yeast strain Kyokai no. 7 (K7) and its relatives, we revealed that several stress-and/or nutrient-responsive transcription factors, particularly Msn2p and Msn4p (Msn2/4p), Hsf1p, Adr1p, and Cat8p, are significantly inactivated (5-7). Impairment of these transcriptional activators in laboratory strains of S. cerevisiae leads to increased fermentation rates, indicating that defective stress responses are linked with the superior fermentation properties of sake yeast (2, 5-7). Moreover, a loss-of-function mutation by insertion of an A residue at position 5055 in the RIM15 gene (rim15 5055insA ) was commonly found among K7-related strains (8). RIM15 encodes a conserved Greatwall-like protein kinase involved in the control of mitot...

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Central carbon metabolism ofSaccharomyces cerevisiaein anaerobic, oxygen-limited and fully aerobic steady-state conditions and following a shift to anaerobic conditions

Cited by 75 publications

References 42 publications

Glucose and Fructose Fermentation by Wine Yeasts in Media Containing Structurally Complex Nitrogen Sources

Glucose and Fructose Fermentation by Wine Yeasts in Media Containing Structurally Complex Nitrogen Sources

Limitations in Xylose-FermentingSaccharomyces cerevisiae, Made Evident through Comprehensive Metabolite Profiling and Thermodynamic Analysis

Inhibitory Role of Greatwall-Like Protein Kinase Rim15p in Alcoholic Fermentation via Upregulating the UDP-Glucose Synthesis Pathway in Saccharomyces cerevisiae

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