scholarly journals Optimizing anaerobic growth rate and fermentation kinetics in Saccharomyces cerevisiae strains expressing Calvin-cycle enzymes for improved ethanol yield

2018 ◽  
Vol 11 (1) ◽  
Author(s):  
Ioannis Papapetridis ◽  
Maaike Goudriaan ◽  
María Vázquez Vitali ◽  
Nikita A. de Keijzer ◽  
Marcel van den Broek ◽  
...  
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Growth Rate ◽  
Ethanol Yield ◽  
Calvin Cycle ◽  
Anaerobic Growth ◽  
Fermentation Kinetics ◽  
Calvin Cycle Enzymes

scholarly journals Oxygen Consumption by Anaerobic Saccharomyces cerevisiae under Enological Conditions: Effect on Fermentation Kinetics

2003 ◽  
Vol 69 (1) ◽  
pp. 113-121 ◽  
Author(s):  
Eric Rosenfeld ◽  
Bertrand Beauvoit ◽  
Bruno Blondin ◽  
Jean-Michel Salmon
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Oxygen Consumption ◽  
Stationary Phase ◽  
Respiratory Chain ◽  
Unsaturated Fatty Acids ◽  
De Novo ◽  
Anaerobic Growth ◽  
Fermentation Rate ◽  
Fermentation Kinetics ◽  
Oxygen Pulse

ABSTRACT The anaerobic growth of the yeast Saccharomyces cerevisiae normally requires the addition of molecular oxygen, which is used to synthesize sterols and unsaturated fatty acids (UFAs). A single oxygen pulse can stimulate enological fermentation, but the biochemical pathways involved in this phenomenon remain to be elucidated. We showed that the addition of oxygen (0.3 to 1.5 mg/g [dry mass] of yeast) to a lipid-depleted medium mainly resulted in the synthesis of the sterols and UFAs required for cell growth. However, the addition of oxygen during the stationary phase in a medium containing excess ergosterol and oleic acid increased the specific fermentation rate, increased cell viability, and shortened the fermentation period. Neither the respiratory chain nor de novo protein synthesis was required for these medium- and long-term effects. As de novo lipid synthesis may be involved in ethanol tolerance, we studied the effect of oxygen addition on sterol and UFA auxotrophs (erg1 and ole1 mutants, respectively). Both mutants exhibited normal anaerobic fermentation kinetics. However, only the ole1 mutant strain responded to the oxygen pulse during the stationary phase, suggesting that de novo sterol synthesis is required for the oxygen-induced increase of the specific fermentation rate. In conclusion, the sterol pathway appears to contribute significantly to the oxygen consumption capacities of cells under anaerobic conditions. Nevertheless, we demonstrated the existence of alternative oxygen consumption pathways that are neither linked to the respiratory chain nor linked to heme, sterol, or UFA synthesis. These pathways dissipate the oxygen added during the stationary phase, without affecting the fermentation kinetics.

scholarly journals ATPase-based implementation of enforced ATP wasting in Saccharomyces cerevisiae for improved ethanol production

2020 ◽  
Vol 13 (1) ◽  
Author(s):  
Ahmed Zahoor ◽  
Katrin Messerschmidt ◽  
Simon Boecker ◽  
Steffen Klamt
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Atpase Activity ◽  
Ethanol Production ◽  
Ethanol Yield ◽  
Volumetric Productivity ◽  
Anaerobic Growth ◽  
Control Strain ◽  
Genomic Integration ◽  
Expression Of Genes ◽  
Atp Generation

Abstract Background Enforced ATP wasting has been recognized as a promising metabolic engineering strategy to enhance the microbial production of metabolites that are coupled to ATP generation. It also appears to be a suitable approach to improve production of ethanol by Saccharomyces cerevisiae. In the present study, we constructed different S. cerevisiae strains with heterologous expression of genes of the ATP-hydrolyzing F1-part of the ATPase enzyme to induce enforced ATP wasting and quantify the resulting effect on biomass and ethanol formation. Results In contrast to genomic integration, we found that episomal expression of the αβγ subunits of the F1-ATPase genes of Escherichia coli in S. cerevisiae resulted in significantly increased ATPase activity, while neither genomic integration nor episomal expression of the β subunit from Trichoderma reesei could enhance ATPase activity. When grown in minimal medium under anaerobic growth-coupled conditions, the strains expressing E. coli’s F1-ATPase genes showed significantly improved ethanol yield (increase of 10% compared to the control strain). However, elevated product formation reduces biomass formation and, therefore, volumetric productivity. We demonstrate that this negative effect can be overcome under growth-decoupled (nitrogen-starved) operation with high and constant biomass concentration. Under these conditions, which mimic the second (production) phase of a two-stage fermentation process, the ATPase-expressing strains showed significant improvement in volumetric productivity (up to 111%) compared to the control strain. Conclusions Our study shows that expression of genes of the F1-portion of E. coli’s ATPase induces ATPase activity in S. cerevisiae and can be a promising way to improve ethanol production. This ATP-wasting strategy can be easily applied to other metabolites of interest, whose formation is coupled to ATP generation.

scholarly journals Properties of yeast grown anaerobically in media limiting in potassium

1971 ◽  
Vol 121 (3) ◽  
pp. 461-467 ◽  
Author(s):  
W. Bartley ◽  
Valerie Broomhead
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Growth Rate ◽  
Yeast Extract ◽  
Maximum Growth ◽  
Anaerobic Growth ◽  
Oxidative Enzymes ◽  
Aerobic Conditions ◽  
Enzyme Pattern ◽  
K Concentration

1. Saccharomyces cerevisiae was grown anaerobically in media with different concentrations of K+ down to less than 1mm. Below 3.2mm the K+ concentration limited the growth rate and yield. 2. Yeast extract was essential for maximum growth. The yield of cells when the medium contained 0.83mm-K+ was only 30% of the yield with 90mm-K+. 3. At the end of anaerobic growth the cells grown in 0.83mm-K+ had a higher concentration of oxidative enzymes than cells grown in 90mm-K+. 4. The cells grown anaerobically in 0.83mm-K+ could adapt to aerobic conditions if K+ was present in the adaptation medium, but not otherwise. 5. The enzyme pattern of the yeast grown aerobically in 0.83mm-K+ was very similar to the anaerobically grown cells and did not change markedly after the glucose was consumed.

scholarly journals Carbon dioxide fixation by Calvin-Cycle enzymes improves ethanol yield in yeast

2013 ◽  
Vol 6 (1) ◽  
pp. 125 ◽  
Author(s):  
Víctor Guadalupe-Medina ◽  
H Wisselink ◽  
Marijke AH Luttik ◽  
Erik de Hulster ◽  
Jean-Marc Daran ◽  
...  
Keyword(s):  
Carbon Dioxide ◽  
Ethanol Yield ◽  
Calvin Cycle ◽  
Carbon Dioxide Fixation ◽  
Calvin Cycle Enzymes

scholarly journals Gpd1 and Gpd2 Fine-Tuning for Sustainable Reduction of Glycerol Formation in Saccharomyces cerevisiae

2011 ◽  
Vol 77 (17) ◽  
pp. 5857-5867 ◽  
Author(s):  
Georg Hubmann ◽  
Stephane Guillouet ◽  
Elke Nevoigt
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Ethanol Yield ◽  
Current Approach ◽  
Fine Tuning ◽  
Anaerobic Growth ◽  
Deletion Strain ◽  
Wild Type ◽  
Double Deletion ◽  
Intermediate Phenotypes ◽  
Glycerol Formation

ABSTRACTGpd1 and Gpd2 are the two isoforms of glycerol 3-phosphate dehydrogenase (GPDH), which is the rate-controlling enzyme of glycerol formation inSaccharomyces cerevisiae.The two isoenzymes play crucial roles in osmoregulation and redox balancing. Past approaches to increase ethanol yield at the cost of reduced glycerol yield have most often been based on deletion of either one or two isogenes (GPD1andGPD2). While single deletions ofGPD1orGPD2reduced glycerol formation only slightly, thegpd1Δgpd2Δ double deletion strain produced zero glycerol but showed an osmosensitive phenotype and abolished anaerobic growth. Our current approach has sought to generate “intermediate” phenotypes by reducing both isoenzyme activities without abolishing them. To this end, theGPD1promoter was replaced in agpd2Δ background by two lower-strengthTEF1promoter mutants. In the same manner, the activity of theGPD2promoter was reduced in agpd1Δbackground. The resulting strains were crossed to obtain different combinations of residualGPD1andGPD2expression levels. Among our engineered strains we identified four candidates showing improved ethanol yields compared to the wild type. In contrast to agpd1Δgpd2Δ double-deletion strain, these strains were able to completely ferment the sugars under quasi-anaerobic conditions in both minimal medium and during simultaneous saccharification and fermentation (SSF) of liquefied wheat mash (wheat liquefact). This result implies that our strains can tolerate the ethanol concentration at the end of the wheat liquefact SSF (up to 90 g liter−1). Moreover, a few of these strains showed no significant reduction in osmotic stress tolerance compared to the wild type.

scholarly journals Engineering of Saccharomyces cerevisiae for Efficient Anaerobic Alcoholic Fermentation of l-Arabinose

2007 ◽  
Vol 73 (15) ◽  
pp. 4881-4891 ◽  
Author(s):  
H. Wouter Wisselink ◽  
Maurice J. Toirkens ◽  
M. del Rosario Franco Berriel ◽  
Aaron A. Winkler ◽  
Johannes P. van Dijken ◽  
...  
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Ethanol Production ◽  
Alcoholic Fermentation ◽  
Plant Biomass ◽  
Ethanol Yield ◽  
Dry Weight ◽  
Pentose Phosphate ◽  
Anaerobic Growth ◽  
Evolutionary Engineering ◽  
First Case

ABSTRACT For cost-effective and efficient ethanol production from lignocellulosic fractions of plant biomass, the conversion of not only major constituents, such as glucose and xylose, but also less predominant sugars, such as l-arabinose, is required. Wild-type strains of Saccharomyces cerevisiae, the organism used in industrial ethanol production, cannot ferment xylose and arabinose. Although metabolic and evolutionary engineering has enabled the efficient alcoholic fermentation of xylose under anaerobic conditions, the conversion of l-arabinose into ethanol by engineered S. cerevisiae strains has previously been demonstrated only under oxygen-limited conditions. This study reports the first case of fast and efficient anaerobic alcoholic fermentation of l-arabinose by an engineered S. cerevisiae strain. This fermentation was achieved by combining the expression of the structural genes for the l-arabinose utilization pathway of Lactobacillus plantarum, the overexpression of the S. cerevisiae genes encoding the enzymes of the nonoxidative pentose phosphate pathway, and extensive evolutionary engineering. The resulting S. cerevisiae strain exhibited high rates of arabinose consumption (0.70 g h−1 g [dry weight]−1) and ethanol production (0.29 g h−1 g [dry weight]−1) and a high ethanol yield (0.43 g g−1) during anaerobic growth on l-arabinose as the sole carbon source. In addition, efficient ethanol production from sugar mixtures containing glucose and arabinose, which is crucial for application in industrial ethanol production, was achieved.

scholarly journals Saccharomyces cerevisiae Fermentation of 28 Barley and 12 Oat Cultivars

Fermentation ◽  
2021 ◽  
Vol 7 (2) ◽  
pp. 59
Author(s):  
Timothy J. Tse ◽  
Daniel J. Wiens ◽  
Jianheng Shen ◽  
Aaron D. Beattie ◽  
Martin J. T. Reaney
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Acetic Acid ◽  
Lactic Acid ◽  
Succinic Acid ◽  
Alcoholic Fermentation ◽  
Ethanol Yield ◽  
Cereal Crops ◽  
Fermentation Products ◽  
Barley Cultivars ◽  
Oat Cultivars

As barley and oat production have recently increased in Canada, it has become prudent to investigate these cereal crops as potential feedstocks for alcoholic fermentation. Ethanol and other coproduct yields can vary substantially among fermented feedstocks, which currently consist primarily of wheat and corn. In this study, the liquified mash of milled grains from 28 barley (hulled and hull-less) and 12 oat cultivars were fermented with Saccharomyces cerevisiae to determine concentrations of fermentation products (ethanol, isopropanol, acetic acid, lactic acid, succinic acid, α-glycerylphosphorylcholine (α-GPC), and glycerol). On average, the fermentation of barley produced significantly higher amounts of ethanol, isopropanol, acetic acid, succinic acid, α-GPC, and glycerol than that of oats. The best performing barley cultivars were able to produce up to 78.48 g/L (CDC Clear) ethanol and 1.81 g/L α-GPC (CDC Cowboy). Furthermore, the presence of milled hulls did not impact ethanol yield amongst barley cultivars. Due to its superior ethanol yield compared to oats, barley is a suitable feedstock for ethanol production. In addition, the accumulation of α-GPC could add considerable value to the fermentation of these cereal crops.

scholarly journals Regulatory Mechanisms Controlling Expression of the DAN/TIR Mannoprotein Genes During Anaerobic Remodeling of the Cell Wall in Saccharomyces cerevisiae

Genetics ◽  
2001 ◽  
Vol 157 (3) ◽  
pp. 1169-1177
Author(s):  
Natalia E Abramova ◽  
Brian D Cohen ◽  
Odeniel Sertil ◽  
Rachna Kapoor ◽  
Kelvin J A Davies ◽  
...  
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Consensus Sequence ◽  
Ectopic Expression ◽  
Constitutive Expression ◽  
Anaerobic Growth ◽  
Zinc Cluster ◽  
Sterol Transport ◽  
Genes Encoding ◽  
Altered Regulation ◽  
Repression Domain

Abstract The DAN/TIR genes of Saccharomyces cerevisiae encode homologous mannoproteins, some of which are essential for anaerobic growth. Expression of these genes is induced during anaerobiosis and in some cases during cold shock. We show that several heme-responsive mechanisms combine to regulate DAN/TIR gene expression. The first mechanism employs two repression factors, Mox1 and Mox2, and an activation factor, Mox4 (for mannoprotein regulation by oxygen). The genes encoding these proteins were identified by selecting for recessive mutants with altered regulation of a dan1::ura3 fusion. MOX4 is identical to UPC2, encoding a binucleate zinc cluster protein controlling expression of an anaerobic sterol transport system. Mox4/Upc2 is required for expression of all the DAN/TIR genes. It appears to act through a consensus sequence termed the AR1 site, as does Mox2. The noninducible mox4Δ allele was epistatic to the constitutive mox1 and mox2 mutations, suggesting that Mox1 and Mox2 modulate activation by Mox4 in a heme-dependent fashion. Mutations in a putative repression domain in Mox4 caused constitutive expression of the DAN/TIR genes, indicating a role for this domain in heme repression. MOX4 expression is induced both in anaerobic and cold-shocked cells, so heme may also regulate DAN/TIR expression through inhibition of expression of MOX4. Indeed, ectopic expression of MOX4 in aerobic cells resulted in partially constitutive expression of DAN1. Heme also regulates expression of some of the DAN/TIR genes through the Rox7 repressor, which also controls expression of the hypoxic gene ANB1. In addition Rox1, another heme-responsive repressor, and the global repressors Tup1 and Ssn6 are also required for full aerobic repression of these genes.

scholarly journals Maintenance of Mitochondrial Morphology Is Linked to Maintenance of the Mitochondrial Genome in Saccharomyces cerevisiae

Genetics ◽  
2002 ◽  
Vol 162 (3) ◽  
pp. 1147-1156 ◽  
Author(s):  
Theodor Hanekamp ◽  
Mary K Thorsness ◽  
Indrani Rebbapragada ◽  
Elizabeth M Fisher ◽  
Corrine Seebart ◽  
...  
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Growth Rate ◽  
Mitochondrial Dna ◽  
Carbon Sources ◽  
Mitochondrial Morphology ◽  
Yeast Saccharomyces Cerevisiae ◽  
Slow Growth Rate ◽  
Dna Migration ◽  
Mitochondrial Dna Stability ◽  
Extragenic Suppressors

Abstract In the yeast Saccharomyces cerevisiae, certain mutant alleles of YME4, YME6, and MDM10 cause an increased rate of mitochondrial DNA migration to the nucleus, carbon-source-dependent alterations in mitochondrial morphology, and increased rates of mitochondrial DNA loss. While single mutants grow on media requiring mitochondrial respiration, any pairwise combination of these mutations causes a respiratory-deficient phenotype. This double-mutant phenotype allowed cloning of YME6, which is identical to MMM1 and encodes an outer mitochondrial membrane protein essential for maintaining normal mitochondrial morphology. Yeast strains bearing null mutations of MMM1 have altered mitochondrial morphology and a slow growth rate on all carbon sources and quantitatively lack mitochondrial DNA. Extragenic suppressors of MMM1 deletion mutants partially restore mitochondrial morphology to the wild-type state and have a corresponding increase in growth rate and mitochondrial DNA stability. A dominant suppressor also suppresses the phenotypes caused by a point mutation in MMM1, as well as by specific mutations in YME4 and MDM10.

scholarly journals Effects of Ultrasound on Fermentation of Glucose to Ethanol by Saccharomyces cerevisiae

Fermentation ◽  
2019 ◽  
Vol 5 (1) ◽  
pp. 16 ◽  
Author(s):  
Luis Huezo ◽  
Ajay Shah ◽  
Frederick Michel
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Mass Transfer ◽  
Glucose Uptake ◽  
Ethanol Production ◽  
Ethanol Yield ◽  
Biofuel Production ◽  
Inhibitory Effects ◽  
Negative Effects ◽  
Intraparticle Mass Transfer ◽  
Improve Mass Transfer

Previous studies have shown that pretreatment of corn slurries using ultrasound improves starch release and ethanol yield during biofuel production. However, studies on its effects on the mass transfer of substrates and products during fermentation have shown that it can have both beneficial and inhibitory effects. In this study, the effects of ultrasound on mass transfer limitations during fermentation were examined. Calculation of the external and intraparticle observable moduli under a range of conditions indicate that no external or intraparticle mass transfer limitations should exist for the mass transfer of glucose, ethanol, or carbon dioxide. Fermentations of glucose to ethanol using Saccharomyces cerevisiae were conducted at different ultrasound intensities to examine its effects on glucose uptake, ethanol production, and yeast population and viability. Four treatments were compared: direct ultrasound at intensities of 23 and 32 W/L, indirect ultrasound (1.4 W/L), and no-ultrasound. Direct and indirect ultrasound had negative effects on yeast performance and viability, and reduced the rates of glucose uptake and ethanol production. These results indicate that ultrasound during fermentation, at the levels applied, is inhibitory and not expected to improve mass transfer limitations.

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