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Starvation Conditions (starvation + condition)
Selected AbstractsAnalysis of the Activated Sludge Process in an MBR under Starvation ConditionsCHEMICAL ENGINEERING & TECHNOLOGY (CET), Issue 3 2006M. Vukovic Abstract An aerobic membrane bioreactor (MBR) at complete biomass retention was studied over a period of time under starvation conditions. Kinetic parameters were determined in a no-feed batch test. The decay rate of activated sludge, kd = 0.05,d,1, was determined by tracking the decrease of MLSS. The ratio of MLVSS/MLSS was in the range 0.76,0.85. The pH values were between 7.02 and 8.23. As a function of different initial concentrations of MLSS, specific nitrification rates qN, decreased from 4.23 to 0.02,mg-N/(g,MLVSS,d) and specific biodegradation rates qb increased from 0.23 to 1.90,mg-COD/(g,MLVSS,d). From experimental data the kinetic constants for respiration, which followed Monod kinetics, were determined as qO2max = 9.8,mg-O2/(g,MLVSS,h), Kx = 2.9,g/dm3. Additionally, a linear correlation between MLSS and mean floc size was found to exist during the biodegradation process. [source] Polarization characteristics and property distributions of a proton exchange membrane fuel cell under cathode starvation conditionsINTERNATIONAL JOURNAL OF ENERGY RESEARCH, Issue 10 2010Dongsoo Ko Abstract Property distribution and polarization characteristics of a proton exchange membrane fuel cell (PEMFC) under cathode starvation conditions were investigated numerically and experimentally for a unit cell. The polarization curves of a lab-scale PEMFC were measured with increasing current density for different cell temperatures (40°C, 50°C, and 60°C) at a relative humidity of 100%. To investigate the local temperature, water content and current density on the membrane, and gas velocity in the channel of the PEMFC, numerical studies using the es-pemfc module of the commercial flow solver STAR-CD, which were matched with experimental data, were conducted. Temperature, current density on the membrane, and water content in the MEA were examined to investigate the effect of cell temperature on performance under the cathode starvation condition. At cathode starvation conditions, the performance of a higher cell temperature condition might drop significantly and the mean temperature on the membrane increase abruptly with increasing cell temperature or current density. Copyright © 2009 John Wiley & Sons, Ltd. [source] Molecular analysis of the phosphorus starvation response in Trichodesmium spp.ENVIRONMENTAL MICROBIOLOGY, Issue 9 2009Elizabeth D. Orchard Summary The marine diazotroph Trichodesmium is a major contributor to primary production and nitrogen fixation in the tropical and subtropical oceans. These regions are often characterized by low phosphorus (P) concentrations, and P starvation of Trichodesmium could limit growth, and potentially constrain nitrogen fixation. To better understand how this genus responds to P starvation we examined four genes involved in P acquisition: two copies of a high-affinity phosphate binding protein (pstS and sphX) and two putative alkaline phosphatases (phoA and phoX). Sequence analysis of these genes among cultured species of Trichodesmium (T. tenue, T. erythraeum, T. thiebautii and T. spiralis) showed that they all are present and conserved within the genus. In T. erythraeum IMS101, the expression of sphX, phoA and phoX were sensitive to P supply whereas pstS was not. The induction of alkaline phosphatase activity corresponded with phoA and phoX expression, but enzyme activity persisted after the expression of these genes returned to basal levels. Additionally, nifH (nitrogenase reductase; involved in nitrogen fixation) expression was downregulated under P starvation conditions. These data highlight molecular level responses to low P and lay a foundation for better understanding the dynamics of Trichodesmium P physiology in low-P environments. [source] Starvation-induced changes in the cell surface of Azospirillum lipoferumFEMS MICROBIOLOGY ECOLOGY, Issue 1 2000Thelma Castellanos Abstract Three starvation regimes (a deficient culture medium, a saline buffer solution and distilled water) were evaluated for their possible effect on cell surface characteristics of Azospirillum lipoferum 1842 related to the initial adsorption of the bacterium to surfaces. The bacteria survived for 7 days in all media although they did not multiply. Upon transfer from a rich growth medium (nutrient agar) to starvation conditions, cell surface hydrophobicity dropped sharply but recovered its initial value within 24 to 48 h, except in phosphate-buffered saline, the length of the recovery period depending on the starvation medium. Starvation affected the sugar affinity of the A. lipoferum cell surface mainly towards p -aminophenyl-,- D -mannopyranoside, to a lesser extent to glucose, but not to other monosaccharides tested. Starvation changed the concentration of several cell surface proteins but did not induce the synthesis of new ones. The cell surface hydrophobic protein (43 kDa) of A. lipoferum 1842 was unaffected by any starvation treatment for a period of up to 48 h, but later disappeared. These data showed that starvation is not a major factor in inducing changes in the cell surface which lead to the primary phase of attachment of Azospirillum to surfaces. [source] Nutrition influences growth and virulence of the insect-pathogenic fungus Metarhizium anisopliaeFEMS MICROBIOLOGY LETTERS, Issue 2 2005Farooq A. Shah Abstract Nutrition influenced growth, sporulation and virulence of the insect pathogenic fungus, Metarhizium anisopliae. Virulent conidia were produced on susceptible insect hosts, 1% yeast extract, 2% peptone, osmotic stress medium (OSM) and CN 10:1 medium. Several strain independent markers were identified that could be used to predict the virulence of M. anisopliae conidia. Virulent conidia typically had high levels of spore bound Pr1, an important cuticle degrading protease, and high germination rates. We also show for the first time that virulent conidia have an endogenous CN ratio below 5.2:1. Real Time PCR revealed that virulent conidia from insects contained significantly higher levels of transcripts of pr1 A and other pathogenicity-related genes than inoculum from artificial media. Of the artificial media studied, 1% yeast extract medium yielded the most virulent conidia, these had higher levels of transcripts of these pathogenicity-related genes than the least virulent conidia from the high conidia yielding CN 35:1 medium (= SDA), however, the levels were significantly lower than those in insect-derived conidia. Our study shows for the first time that the passaged inoculum is virulent irrespective of the original culture medium or insect host. Virulent conidia were consistently produced on OSM even though growth and sporulation were poor. We postulate that starvation conditions, whether in vivo or in vitro, results in de-repression of Pr1 and that elevated levels of this enzyme enhance fungal virulence. [source] Role of laccase in the biology and virulence of Cryptococcus neoformansFEMS YEAST RESEARCH, Issue 1 2004Xudong Zhu Abstract Laccase is an important virulence factor for the human pathogen, Cryptococcus neoformans. In this review, we examine the structural, biological and genetic features of the enzyme and its role in the pathogenesis of cryptococcosis. Laccase is expressed in C. neoformans as a cell wall enzyme that possesses a broad spectrum of activity oxidizing both polyphenolic compounds and iron. Two paralogs, CNLAC1 and CNLAC2, are present in the fungus, of which the first one expresses the dominant enzyme activity under glucose starvation conditions. Regulation of the enzyme is in response to various environmental signals including nutrient starvation, the presence of multivalent cations and temperature stress, and is mediated through multiple signal transduction pathways. Study of the function and regulation of this important virulence factor has led to further understanding of mechanisms of fungal pathogenesis and the regulation of stress response in the host cell environment. [source] Transport of phosphatidylinositol 3-phosphate into the vacuole via autophagic membranes in Saccharomyces cerevisiaeGENES TO CELLS, Issue 6 2008Keisuke Obara Vps34, the sole PtdIns 3-kinase in yeast, is essential for autophagy. Here, we show that the lipid-kinase activity of Vps34 is required for autophagy, implying an essential role of its product PtdIns(3)P. The protein-kinase activity of Vps15, a regulatory subunit of the PtdIns 3-kinase complex, is also required for efficient autophagy. We monitored the distribution of PtdIns(3)P in living cells using a specific indicator, the 2xFYVE domain derived from mammalian Hrs. PtdIns(3)P was abundant at endosomes and on the vacuolar membrane during logarithmic growth phase. Under starvation conditions, we observed massive transport of PtdIns(3)P into the vacuole. This accumulation was dependent on the membrane dynamics of autophagy. Notably, PtdIns(3)P was highly enriched and delivered into the vacuole as a component of autophagosome membranes but not as a cargo enclosed within them, implying direct involvement of this phosphoinositide in autophagosome formation. We also found a possible enrichment of PtdIns(3)P on the inner autophagosomal membrane compared to the outer membrane. Based on these results we discuss the function of PtdIns(3)P in autophagy. [source] Phosphatidylinositol 3-phosphate 5-kinase is required for the cellular response to nutritional starvation and mating pheromone signals in Schizosaccharomyces pombeGENES TO CELLS, Issue 2 2002Masayo Morishita Background: Phosphatidylinositol (3,5) bisphosphate, which is converted from phosphatidylinositol 3-phosphate by phosphatidylinositol 3-phosphate 5-kinase, is implicated in vacuolar functions and the sorting of cell surface proteins within endosomes in the endocytic pathway of budding yeast. A homologous protein, SpFab1p, has been found in the fission yeast Schizosaccharomyces pombe, but its role is not known. Results: Here we report that SpFab1p is encoded by ste12+ known as a fertility gene in S. pombe. The ste12 mutant grew normally under stress-free conditions, but was highly vacuolated and swelled at high temperatures and under starvation conditions. In nitrogen-free medium, ste12 cells were arrested in G1 phase, but partially defective in the expression of genes responsible for mating and meiosis. The ste12 mutant was defective both in the production of, and in the response to, mating pheromones. The amount of the pheromone receptor protein Map3p, was substantially decreased in ste12 cells. Map3p was transported to the cell surface, then internalized and eventually transported to the vacuolar lumen, even in the ste12 mutant. Conclusion: The results indicate that phosphatidylinositol(3,5)bisphosphate is essential for cellular responses to various stresses and for the mating pheromone signalling under starvation conditions. [source] Polarization characteristics and property distributions of a proton exchange membrane fuel cell under cathode starvation conditionsINTERNATIONAL JOURNAL OF ENERGY RESEARCH, Issue 10 2010Dongsoo Ko Abstract Property distribution and polarization characteristics of a proton exchange membrane fuel cell (PEMFC) under cathode starvation conditions were investigated numerically and experimentally for a unit cell. The polarization curves of a lab-scale PEMFC were measured with increasing current density for different cell temperatures (40°C, 50°C, and 60°C) at a relative humidity of 100%. To investigate the local temperature, water content and current density on the membrane, and gas velocity in the channel of the PEMFC, numerical studies using the es-pemfc module of the commercial flow solver STAR-CD, which were matched with experimental data, were conducted. Temperature, current density on the membrane, and water content in the MEA were examined to investigate the effect of cell temperature on performance under the cathode starvation condition. At cathode starvation conditions, the performance of a higher cell temperature condition might drop significantly and the mean temperature on the membrane increase abruptly with increasing cell temperature or current density. Copyright © 2009 John Wiley & Sons, Ltd. [source] Autolytic phenotype of Lactococcus lactis strains isolated from traditional Tunisian dairy productsJOURNAL OF APPLIED MICROBIOLOGY, Issue 5 2002H. Ouzari Aims:,To evaluate the autolytic properties of Lactococcus lactis strains isolated from artisan Tunisian dairy products, their peptidoglycan hydrolase content and their activity spectrum. Methods and Results:,The autolytic phenotype of Lactococcus strains was evaluated under starvation conditions in potassium phosphate buffer. The results obtained highlighted a high degree of diversity among the strains analysed, allowing the identification of high and low autolytic Lactococcus lactis strains. Peptidoglycan hydrolase content was evaluated by renaturing SDS-PAGE using cells of Micrococcus lysodeikticus as a target for the enzymatic activity. A major activity band migrating at about 45 kDa was observed. The lytic activity, evaluated in the presence of different chemicals, was retained in 8% NaCl, 15 mmol l,1 CaCl2, and in a pH range between 5 and 9·5. The substrate specificity of peptidoglycan hydrolase from Lactococcus strains was evaluated in renaturing SDS-PAGE incorporating cells of different bacterial species. The major autolysin of Lactococcus lactis was active against cells of Lactococcus lactis subsp. lactis, Streptococcus thermophilus, Lactobacillus delbrueckii subsp. bulgaricus, Lactobacillus helveticus and Listeria monocytogenes. Conclusions:,Autolytic activity is widely distributed in Lactococcus lactis and the rate of autolysis is strain-dependent. The major peptidoglycan hydrolase showed a wide spectrum of activity against several lactic acid bacteria and bacterial species involved in food-related infection. Significance and Impact of the Study:,The autolytic phenotype of Lactococcus lactis strains isolated from Tunisian artisan dairy products has been determined, and the data obtained should allow the selection of strains of technological interest in the cheese-ripening process. [source] Cleavage of mRNAs and role of tmRNA system under amino acid starvation in Escherichia coliMOLECULAR MICROBIOLOGY, Issue 2 2008Xia Li Summary We have shown previously that ribosome stalling during translation caused by various reasons leads to mRNA cleavage, resulting in non-stop mRNAs that are eliminated in a tmRNA-dependent manner. Amino acid starvation is a physiological condition in which ribosome stalling is expected to occur more frequently. Here we demonstrate that mRNA cleavage is induced by amino acid starvation, resulting in accumulation of truncated mRNAs in cells lacking tmRNA. The truncated mRNAs are eliminated in wild-type cells, indicating that the tmRNA system rapidly degrade the truncated mRNAs. The cleavage pattern of model mRNAs in which serine codons were replaced with threonine codons indicated that mRNA cleavage occurs near serine codons in response to serine starvation. Cells lacking all of the five known toxin loci were proficient in mRNA cleavage, showing that toxin,antitoxin systems are not responsible for the cleavage. A mild serine starvation caused a significant growth inhibition in cells lacking tmRNA but not in wild-type cells. The ribosome-mediated mRNA cleavage along with the tmRNA system is an important mechanism that enables cells to adapt to amino acid starvation conditions. [source] Non-growing Escherichia coli cells starved for glucose or phosphate use different mechanisms to survive oxidative stress,MOLECULAR MICROBIOLOGY, Issue 4 2001Patrice L. Moreau Recent data suggest that superoxide dismutases are important in preventing lethal oxidative damage of proteins in Escherichia coli cells incubated under aerobic, carbon starvation conditions. Here, we show that the alkylhydroperoxide reductase AhpCF (AHP) is specifically required to protect cells incubated under aerobic, phosphate (Pi) starvation conditions. Additional loss of the HP-I (KatG) hydroperoxidase activity dramatically accelerated the death rate of AHP-deficient cells. Investigation of the composition of spent culture media indicates that ,ahpCF katG cells leak nutrients, which suggests that membrane lipids are the principal target of peroxides produced in Pi-starved cells. In fact, the introduction of various mutations inactivating repair activities revealed no obvious role for protein or DNA lesions in the viability of ahp cells. Because the death of ahp cells was directly related to ongoing aerobic glucose metabolism, we wondered how glycolysis, which requires free Pi, could proceed. 31P nuclear magnetic resonance spectra showed that Pi-starved cells consumed Pi but were apparently able to liberate Pi from phosphorylated products, notably through the synthesis of UDP-glucose. Whereas expression of the ahpCF and katG genes is enhanced in an OxyR-dependent manner in response to H2O2 challenge, we found that the inactivation of oxyR and both oxyR and rpoS genes had little effect on the viability of Pi-starved cells. In stark contrast, the inactivation of both oxyR and rpoS genes dramatically decreased the viability of glucose-starved cells. [source] Proteolysis during long-term glucose starvation in Staphylococcus aureus COLPROTEINS: STRUCTURE, FUNCTION AND BIOINFORMATICS, Issue 19 2009Stephan Michalik Abstract A combination of pulse-chase experiments and 2-D PAGE revealed that protein degradation appears to play a crucial role for the cell physiology of Staphylococcus aureus COL during extended periods of glucose starvation. The synthesis rate of virtually all cytosolic and radioactively labeled proteins from growing cells seemed dramatically reduced in the first 3.5,h of glucose starvation. The stability of proteins synthesized in growing cells was monitored by a pulse-chase approach on a proteome wide scale. Especially, enzymes involved in nucleic acid and amino acid biosyntheses, energy metabolism and biosynthesis of cofactors were found rather rapidly degraded within the onset of the stationary phase, whereas the majority of glycolytic and tricarboxylic acid cycle enzymes remained more stable. Furthermore, single enzymes of biosynthetic pathways were differentially degraded. A metabolite analysis revealed that glucose completely depleted from the medium in the transient phase, and amino acids such as alanine and glycine were taken up by the cells in the stationary phase. We suggest that vegetative proteins no longer required in non-growing cells and thus no longer protected by integration into functional complexes were degraded. Proteolysis of putative non-substrate-bound or "unemployed" proteins appears to be a characteristic feature of S. aureus in order to access nutrients as an important survival strategy under starvation conditions. [source] The glucose and nitrogen starvation response of Bacillus licheniformisPROTEINS: STRUCTURE, FUNCTION AND BIOINFORMATICS, Issue 3 2007Birgit Voigt Abstract The glucose and nitrogen starvation stimulons of Bacillus licheniformis were determined by transcriptome and proteome analyses. Under both starvation conditions, the main response of B. licheniformis was a switch to the usage of alternative nutrient sources. This was indicated by an induction of genes involved in the metabolism of C-2 substrates during glucose limitation. In addition, B. licheniformis seems to be using other organic substances like amino acids and lipids as carbon sources when subjected to glucose starvation. This observation is supported by the induction of a high number of genes coding for proteins involved in amino acid and lipid degradation. During nitrogen starvation, genes for several proteases and peptidases involved in nitrate and nitrite assimilation were induced, which enables this bacterium to recruit nitrogen from alternative sources. Both starvation conditions led to a down-regulation of transcription of most vegetative genes, which was subsequently reflected by a reduced synthesis of the corresponding proteins. A selected set of genes was induced by both starvation conditions. Among them were yvyD, citA and the putative methylcitrate shunt genes mmgD, mmgE and yqiQ. However, both starvation conditions did not induce a general SigmaB-dependent stress response. [source] The extracellular proteome of Bacillus licheniformis grown in different media and under different nutrient starvation conditionsPROTEINS: STRUCTURE, FUNCTION AND BIOINFORMATICS, Issue 5 2006Birgit Voigt Dr. No abstracts. [source] Control of misincorporation of serine for asparagine during antibody production using CHO cellsBIOTECHNOLOGY & BIOENGINEERING, Issue 1 2010Anurag Khetan Abstract A recombinant monoclonal antibody produced by Chinese hamster ovary (CHO) cell fed-batch culture was found to have amino acid sequence misincorporation upon analysis by intact mass and peptide mapping mass spectrometry. A detailed analysis revealed multiple sites for asparagine were being randomly substituted by serine, pointing to mistranslation as the likely source. Results from time-course analysis of cell culture suggest that misincorporation was occurring midway through the fed-batch process and was correlated to asparagine reduction to below detectable levels in the culture. Separate shake flask experiments were carried out that confirmed starvation of asparagine and not excess of serine in the medium as the root cause of the phenomenon. Reduction in serine concentration under asparagine starvation conditions helped reduce extent of misincorporation. Supplementation with glutamine also helped reduce extent of misincorporation. Maintenance of asparagine at low levels in 2,L bench-scale culture via controlled supplementation of asparagine-containing feed eliminated the occurrence of misincorporation. This strategy was implemented in a clinical manufacturing process and scaled up successfully to the 200 and 2,000,L bioreactor scales. Biotechnol. Bioeng. 2010;107: 116,123. © 2010 Wiley Periodicals, Inc. [source] BAK and BAX deletion using zinc-finger nucleases yields apoptosis-resistant CHO cellsBIOTECHNOLOGY & BIOENGINEERING, Issue 2 2010Gregory J. Cost Abstract Anoxic and metabolic stresses in large-scale cell culture during biopharmaceutical production can induce apoptosis. Strategies designed to ameliorate the problem of apoptosis in cell culture have focused on mRNA knockdown of pro-apoptotic proteins and over-expression of anti-apoptotic ones. Apoptosis in cell culture involves mitochondrial permeabilization by the pro-apoptotic Bak and Bax proteins; activity of either protein is sufficient to permit apoptosis. We demonstrate here the complete and permanent elimination of both the Bak and Bax proteins in combination in Chinese hamster ovary (CHO) cells using zinc-finger nuclease-mediated gene disruption. Zinc-finger nuclease cleavage of BAX and BAK followed by inaccurate DNA repair resulted in knockout of both genes. Cells lacking Bax and Bak grow normally but fail to activate caspases in response to apoptotic stimuli. When grown using scale-down systems under conditions that mimic growth in large-scale bioreactors they are significantly more resistant to apoptosis induced by starvation, staurosporine, and sodium butyrate. When grown under starvation conditions, BAX - and BAK -deleted cells produce two- to fivefold more IgG than wild-type CHO cells. Under normal growth conditions in suspension culture in shake flasks, double-knockout cultures achieve equal or higher cell densities than unmodified wild-type cultures and reach viable cell densities relevant for large-scale industrial protein production. Biotechnol. Bioeng. 2010; 105: 330,340. © 2009 Wiley Periodicals, Inc. [source] Analysis of the Activated Sludge Process in an MBR under Starvation ConditionsCHEMICAL ENGINEERING & TECHNOLOGY (CET), Issue 3 2006M. Vukovic Abstract An aerobic membrane bioreactor (MBR) at complete biomass retention was studied over a period of time under starvation conditions. Kinetic parameters were determined in a no-feed batch test. The decay rate of activated sludge, kd = 0.05,d,1, was determined by tracking the decrease of MLSS. The ratio of MLVSS/MLSS was in the range 0.76,0.85. The pH values were between 7.02 and 8.23. As a function of different initial concentrations of MLSS, specific nitrification rates qN, decreased from 4.23 to 0.02,mg-N/(g,MLVSS,d) and specific biodegradation rates qb increased from 0.23 to 1.90,mg-COD/(g,MLVSS,d). From experimental data the kinetic constants for respiration, which followed Monod kinetics, were determined as qO2max = 9.8,mg-O2/(g,MLVSS,h), Kx = 2.9,g/dm3. Additionally, a linear correlation between MLSS and mean floc size was found to exist during the biodegradation process. [source] |