scholarly journals Identity of the Growth-Limiting Nutrient Strongly Affects Storage Carbohydrate Accumulation in Anaerobic Chemostat Cultures of Saccharomyces cerevisiae

2009 ◽  
Vol 75 (21) ◽  
pp. 6876-6885 ◽  
Author(s):  
Lucie A. Hazelwood ◽  
Michael C. Walsh ◽  
Marijke A. H. Luttik ◽  
Pascale Daran-Lapujade ◽  
Jack T. Pronk ◽  
...  
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Growth Rate ◽  
Glycogen Phosphorylase ◽  
Specific Growth ◽  
Glycogen Synthase ◽  
The Other ◽  
Carbohydrate Accumulation ◽  
Storage Carbohydrate ◽  
Chemostat Cultures ◽  
Limiting Nutrient

ABSTRACT Accumulation of glycogen and trehalose in nutrient-limited cultures of Saccharomyces cerevisiae is negatively correlated with the specific growth rate. Additionally, glucose-excess conditions (i.e., growth limitation by nutrients other than glucose) are often implicated in high-level accumulation of these storage carbohydrates. The present study investigates how the identity of the growth-limiting nutrient affects accumulation of storage carbohydrates in cultures grown at a fixed specific growth rate. In anaerobic chemostat cultures (dilution rate, 0.10 h−1) of S. cerevisiae, the identity of the growth-limiting nutrient (glucose, ammonia, sulfate, phosphate, or zinc) strongly affected storage carbohydrate accumulation. The glycogen contents of the biomass from glucose- and ammonia-limited cultures were 10- to 14-fold higher than those of the biomass from cultures grown under the other three glucose-excess regimens. Trehalose levels were specifically higher under nitrogen-limited conditions. These results demonstrate that storage carbohydrate accumulation in nutrient-limited cultures of S. cerevisiae is not a generic response to excess glucose but instead is strongly dependent on the identity of the growth-limiting nutrient. While transcriptome analysis of wild-type and msn2Δ msn4Δ strains confirmed that transcriptional upregulation of glycogen and trehalose biosynthesis genes is mediated by Msn2p/Msn4p, transcriptional regulation could not quantitatively account for the drastic changes in storage carbohydrate accumulation. The results of assays of glycogen synthase and glycogen phosphorylase activities supported involvement of posttranscriptional regulation. Consistent with the high glycogen levels in ammonia-limited cultures, the ratio of glycogen synthase to glycogen phosphorylase in these cultures was up to eightfold higher than the ratio in the other glucose-excess cultures.

scholarly journals Control of specific growth rate in Saccharomyces cerevisiae

Microbiology ◽  
2009 ◽  
Vol 155 (5) ◽  
pp. 1699-1707 ◽  
Author(s):  
J. L. Snoep ◽  
M. Mrwebi ◽  
J. M. Schuurmans ◽  
J. M. Rohwer ◽  
M. J. Teixeira de Mattos
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Growth Rate ◽  
Specific Growth Rate ◽  
Specific Growth ◽  
Full Control ◽  
Chemostat Cultures

In this contribution we resolve the long-standing dispute whether or not the Monod constant (KS), describing the overall affinity of an organism for its growth-limiting substrate, can be related to the affinity of the transporter for that substrate (KM). We show how this can be done via the control of the transporter on the specific growth rate; they are identical if the transport step has full control. The analysis leads to the counter-intuitive result that the affinity of an organism for its substrate is expected to be higher than the affinity of the enzyme that facilitates its transport. Experimentally, we show this indeed to be the case for the yeast Saccharomyces cerevisiae, for which we determined a KM value for glucose more than two times higher than the KS value in glucose-limited chemostat cultures. Moreover, we calculated that at glucose concentrations of 0.03 and 0.29 mM, the transport step controls the specific growth rate at 78 and 49 %, respectively.

scholarly journals Analysis of Gene Expression in Escherichia coli in Response to Changes of Growth-Limiting Nutrient in Chemostat Cultures

2004 ◽  
Vol 70 (4) ◽  
pp. 2354-2366 ◽  
Author(s):  
Qiang Hua ◽  
Chen Yang ◽  
Taku Oshima ◽  
Hirotada Mori ◽  
Kazuyuki Shimizu
Keyword(s):  
Gene Expression ◽  
Escherichia Coli ◽  
Growth Rate ◽  
Steady State ◽  
Specific Growth ◽  
Differentially Expressed ◽  
Transcriptional Responses ◽  
Chemostat Cultures ◽  
Limiting Nutrient ◽  
Slow Growing

ABSTRACT Studies of steady-state metabolic fluxes in Escherichia coli grown in nutrient-limited chemostat cultures suggest remarkable flux alterations in response to changes of growth-limiting nutrient in the medium (Hua et al., J. Bacteriol. 185:7053-7067, 2003). To elucidate the physiological adaptation of cells to the nutrient condition through the flux change and understand the molecular mechanisms underlying the change in the flux, information on gene expression is of great importance. DNA microarray analysis was performed to investigate the global transcriptional responses of steady-state cells grown in chemostat cultures with limited glucose or ammonia while other environmental conditions and the growth rate were kept constant. In slow-growing cells (specific growth rate of 0.10 h−1), 9.8% of a total of 4,071 genes investigated, especially those involved in amino acid metabolism, central carbon and energy metabolism, transport system and cell envelope, were observed to be differentially expressed between the two nutrient-limited cultures. One important characteristic of E. coli grown under nutrient limitation was its capacity to scavenge carbon or nitrogen from the medium through elevating the expression of the corresponding transport and assimilation genes. The number of differentially expressed genes in faster-growing cells (specific growth rate of 0.55 h−1), however, decreased to below half of that in slow-growing cells, which could be explained by diverse transcriptional responses to the growth rate under different nutrient limitations. Independent of the growth rate, 92 genes were identified as being differentially expressed. Genes tightly related to the culture conditions were highlighted, some of which may be used to characterize nutrient-limited growth.

scholarly journals Influence of organic acids and organochlorinated insecticides on metabolism of Saccharomyces cerevisiae

2005 ◽  
pp. 207-215 ◽  
Author(s):  
Dusanka Pejin ◽  
Vesna Vasic
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Growth Rate ◽  
Sugar Beet ◽  
Specific Growth Rate ◽  
Glycogen Content ◽  
Specific Growth ◽  
Raw Materials ◽  
Stress Factors ◽  
Adaptive Mechanism ◽  
Ribonucleic Acids

Saccharomyces cerevisiae is exposed to different stress factors during the production: osmotic, temperature, oxidative. The response to these stresses is the adaptive mechanism of cells. The raw materials Saccharomyces cerevisiae is produced from, contain metabolism products of present microorganisms and protective agents used during the growth of sugar beet for example the influence of acetic and butyric acid and organochlorinated insecticides, lindan and heptachlor, on the metabolism of Saccharomyces cerevisiae was investigated and presented in this work. The mentioned compounds affect negatively the specific growth rate, yield, content of proteins, phosphorus, total ribonucleic acids. These compounds influence the increase of trechalose and glycogen content in the Saccharomyces cerevisiae cells.

scholarly journals Adaptive laboratory evolution and reverse engineering of single-vitamin prototrophies in Saccharomyces cerevisiae

2020 ◽  
Author(s):  
Thomas Perli ◽  
Dewi P.I. Moonen ◽  
Marcel van den Broek ◽  
Jack T. Pronk ◽  
Jean-Marc Daran
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Growth Rate ◽  
Nicotinic Acid ◽  
Specific Growth ◽  
Pantothenic Acid ◽  
B Vitamins ◽  
Fast Growth ◽  
Adaptive Laboratory Evolution ◽  
Laboratory Evolution ◽  
B Vitamin

AbstractQuantitative physiological studies on Saccharomyces cerevisiae commonly use synthetic media (SM) that contain a set of water-soluble growth factors that, based on their roles in human nutrition, are referred to as B-vitamins. Previous work demonstrated that, in S. cerevisiae CEN.PK113-7D, requirements for biotin could be eliminated by laboratory evolution. In the present study, this laboratory strain was shown to exhibit suboptimal specific growth rates when either inositol, nicotinic acid, pyridoxine, pantothenic acid, para-aminobenzoic acid (pABA) or thiamine were omitted from SM. Subsequently, this strain was evolved in parallel serial-transfer experiments for fast aerobic growth on glucose in the absence of individual B-vitamins. In all evolution lines, specific growth rates reached at least 90 % of the growth rate observed in SM supplemented with a complete B-vitamin mixture. Fast growth was already observed after a few transfers on SM without myo-inositol, nicotinic acid or pABA. Reaching similar results in SM lacking thiamine, pyridoxine or pantothenate required over 300 generations of selective growth. The genomes of evolved single-colony isolates were re-sequenced and, for each B-vitamin, a subset of non-synonymous mutations associated with fast vitamin-independent growth were selected. These mutations were introduced in a non-evolved reference strain using CRISPR/Cas9-based genome editing. For each B-vitamin, introduction of a small number of mutations sufficed to achieve substantially a increased specific growth rate in non-supplemented SM that represented at least 87% of the specific growth rate observed in fully supplemented complete SM.ImportanceMany strains of Saccharomyces cerevisiae, a popular platform organism in industrial biotechnology, carry the genetic information required for synthesis of biotin, thiamine, pyridoxine, para-aminobenzoic acid, pantothenic acid, nicotinic acid and inositol. However, omission of these B-vitamins typically leads to suboptimal growth. This study demonstrates that, for each individual B-vitamin, it is possible to achieve fast vitamin-independent growth by adaptive laboratory evolution (ALE). Identification of mutations responsible for these fast-growing phenotype by whole-genome sequencing and reverse engineering showed that, for each compound, a small number of mutations sufficed to achieve fast growth in its absence. These results form an important first step towards development of S. cerevisiae strains that exhibit fast growth on cheap, fully mineral media that only require complementation with a carbon source, thereby reducing costs, complexity and contamination risks in industrial yeast fermentation processes.

scholarly journals Anaplerotic reactions active during growth of Saccharomyces cerevisiae on glycerol

2019 ◽  
Vol 20 (1) ◽  
Author(s):  
Joeline Xiberras ◽  
Mathias Klein ◽  
Celina Prosch ◽  
Zahabiya Malubhoy ◽  
Elke Nevoigt
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Growth Rate ◽  
Specific Growth Rate ◽  
Pyruvate Carboxylase ◽  
Reference Strain ◽  
Specific Growth ◽  
Glyoxylate Cycle ◽  
Maximum Specific Growth Rate ◽  
The Glyoxylate Cycle ◽  
Anaplerotic Reactions

ABSTRACT Anaplerotic reactions replenish TCA cycle intermediates during growth. In Saccharomyces cerevisiae, pyruvate carboxylase and the glyoxylate cycle have been experimentally identified to be the main anaplerotic routes during growth on glucose (C6) and ethanol (C2), respectively. The current study investigates the importance of the two isoenzymes of pyruvate carboxylase (PYC1 and PYC2) and one of the key enzymes of the glyoxylate cycle (ICL1) for growth on glycerol (C3) as a sole carbon source. As the wild-type strains of the CEN.PK family are unable to grow in pure synthetic glycerol medium, a reverse engineered derivative showing a maximum specific growth rate of 0.14 h−1 was used as the reference strain. While the deletion of PYC1 reduced the maximum specific growth rate by about 38%, the deletion of PYC2 had no significant impact, neither in the reference strain nor in the pyc1Δ mutant. The deletion of ICL1 only marginally reduced growth of the reference strain but further decreased the growth rate of the pyc1 deletion strain by 20%. Interestingly, the triple deletion (pyc1Δ pyc2Δ icl1Δ) did not show any growth. Therefore, both the pyruvate carboxylase and the glyoxylate cycle are involved in anaplerosis during growth on glycerol.

Die Wirkung eines homogenen Magnetfeldes auf das Wachstum von Micrococcus denitrificans / The Influence of Homogeneous Magnetic Fields on the Growth of Micrococcus denitrificans

1970 ◽  
Vol 25 (9) ◽  
pp. 1020-1023 ◽  
Author(s):  
Wolfram Thiemann ◽  
Erich Wagner
Keyword(s):  
Magnetic Field ◽  
Saccharomyces Cerevisiae ◽  
Growth Rate ◽  
Magnetic Fields ◽  
The Other ◽  
Anaerobic Conditions ◽  
Inhibition Of Growth ◽  
Significant Acceleration ◽  
In The Beginning ◽  
The Magnetic Field

The influence of strong homogeneous magnetic fields in the range of 5000 to 8000 Gauss on the growth of Saccharomyces cerevisiae and Micrococcus denitrificans was studied. In the case of yeast growing under nearly anaerobic conditions an inhibition of growth rate was observed in the beginning of incubaton while some hours later the growth accelerated and surpassed the control. M. denitrificans on the other hand grew with the same rate as the controls during the first 2 - 3 hours of experiment; thereafter the magnetic field resulted in a significant acceleration of growth rate measured by a 5.8 to 13.3% increase of oxygen consumption after 5 - 6 hours run of experiment. Until now only inhibition of bacterial growths by magnetic fields is reported elsewhere in the literature.

scholarly journals Prolonged Maltose-Limited Cultivation of Saccharomyces cerevisiae Selects for Cells with Improved Maltose Affinity and Hypersensitivity

2004 ◽  
Vol 70 (4) ◽  
pp. 1956-1963 ◽  
Author(s):  
Mickel L. A. Jansen ◽  
Pascale Daran-Lapujade ◽  
Johannes H. de Winde ◽  
Matthew D. W. Piper ◽  
Jack T. Pronk
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Substrate Affinity ◽  
Maltose Permease ◽  
Prolonged Cultivation ◽  
Chemostat Cultures ◽  
Proton Symport ◽  
Limiting Nutrient ◽  
Substrate Saturation ◽  
Selection For ◽  
Substrate Tolerance

ABSTRACT Prolonged cultivation (>25 generations) of Saccharomyces cerevisiae in aerobic, maltose-limited chemostat cultures led to profound physiological changes. Maltose hypersensitivity was observed when cells from prolonged cultivations were suddenly exposed to excess maltose. This substrate hypersensitivity was evident from massive cell lysis and loss of viability. During prolonged cultivation at a fixed specific growth rate, the affinity for the growth-limiting nutrient (i.e., maltose) increased, as evident from a decreasing residual maltose concentration. Furthermore, the capacity of maltose-dependent proton uptake increased up to 2.5-fold during prolonged cultivation. Genome-wide transcriptome analysis showed that the increased maltose transport capacity was not primarily due to increased transcript levels of maltose-permease genes upon prolonged cultivation. We propose that selection for improved substrate affinity (ratio of maximum substrate consumption rate and substrate saturation constant) in maltose-limited cultures leads to selection for cells with an increased capacity for maltose uptake. At the same time, the accumulative nature of maltose-proton symport in S. cerevisiae leads to unrestricted uptake when maltose-adapted cells are exposed to a substrate excess. These changes were retained after isolation of individual cell lines from the chemostat cultures and nonselective cultivation, indicating that mutations were involved. The observed trade-off between substrate affinity and substrate tolerance may be relevant for metabolic engineering and strain selection for utilization of substrates that are taken up by proton symport.

Morphogenetic expression of Arthrobacter globiformis 425 in continuous culture with carbon or biotin limitation

1978 ◽  
Vol 24 (1) ◽  
pp. 28-30 ◽  
Author(s):  
Adrian P. Wills ◽  
E. C. S. Chan
Keyword(s):  
Growth Rate ◽  
Growth Rates ◽  
Specific Growth ◽  
Normal Growth ◽  
Arthrobacter Globiformis ◽  
Glucose Limitation ◽  
Abnormal Morphology ◽  
Chemostat Cultures ◽  
Specific Growth Rates ◽  
Biotin Limitation

When deprived of biotin, Arthrobacter globiformis 425 exhibits abnormal morphology (large, branched forms of variable size) and a retardation of its normal growth rate. In chemostat cultures, when cells were grown under glucose limitation, the morphology was normal (coccoids or rods) at specific growth rates between 0.05 and 0.125 h−1 (doubling times between 14 and 5.5 h, respectively) at 25 °C. The coccoid-to-rod morphogenesis occurs at a specific growth rate of 0.11 h−1. At the same specific growth rates and temperature, but under biotin limitation, abnormal morphology was observed.

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