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  • 1
    Digitale Medien
    Digitale Medien
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 153 (1997), S. 0 
    ISSN: 1574-6968
    Quelle: Blackwell Publishing Journal Backfiles 1879-2005
    Thema: Biologie
    Notizen: This work reports the intracellular pH (pHi) dynamics of Saccharomyces cerevisiae cells in sporulation medium. Cells loaded with the pH-sensitive dye carboxy-seminaphthorhodafluor-1 (C.SNARF-1) exhibited an alkalization of the pHi following the extracellular pH during sporulation in the absence of buffer and almost no change in pHi or ΔpH when sporulation was carried out in buffered medium. The results indicate that the pH gradient does not appear to be directly involved in the regulation of acetate uptake during sporulation. However, the alkalization of pHi by eliciting a decrease in metabolic fluxes could account for a lower demand for acetate.
    Materialart: Digitale Medien
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  • 2
    Digitale Medien
    Digitale Medien
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 194 (2001), S. 0 
    ISSN: 1574-6968
    Quelle: Blackwell Publishing Journal Backfiles 1879-2005
    Thema: Biologie
    Notizen: A simple, rapid and reliable procedure for permeabilizing cyanobacterial cells and measuring the glycogen synthetic pathway in situ, is presented. Cells from Anabaena sp. strain PCC 7120 were permeabilized with a mixture of toluene:ethanol (1:4 v/v). Fluorescence microscopy of cells incubated with fluorescein diacetate showed Anabaena non-permeabilized cells as green fluorescents, whereas permeabilized (viable) cells exhibited the intrinsic red fluorescence. Labelled α-1,4-glucan was recovered when permeabilized cells were incubated with the substrates of ADP-glucose pyrophosphorylase or glycogen synthase. The kinetic and regulatory properties of both enzymes could be reproduced in situ. The simplicity of the procedure and the ability to measure in situ glucan fluxes show the methodology as useful for studying the intracellular regulation of storage polysaccharides in a photosynthetic prokaryote.
    Materialart: Digitale Medien
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  • 3
    Digitale Medien
    Digitale Medien
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 66 (1990), S. 0 
    ISSN: 1574-6968
    Quelle: Blackwell Publishing Journal Backfiles 1879-2005
    Thema: Biologie
    Notizen: Abstract A stoichiometric model of anaerobic glycolysis is presented and the influence on its dynamics by the ATP-consuming membrane transport processes and substrate input rate are studied. The model is represented by a system of four ODE (ordinary differential equations), mass conservation equations and functions of state variables, such as thermodynamic efficiency. A low substrate input rate provokes damped oscillations while a high enrgy load determines sustained oscillations in all the metabolites and in thermodynamic efficiency. Due to the lack of linearity between fluxes and forces in the oscillatory region it may be stated that oscillations appear when the system is kinetically controlled.
    Materialart: Digitale Medien
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  • 4
    Digitale Medien
    Digitale Medien
    Springer
    Molecular and cellular biochemistry 126 (1993), S. 115-124 
    ISSN: 1573-4919
    Schlagwort(e): DNA supercoiling ; DNA topoisomerases ; [ATP]/[ADP] ratio ; aerobic anaerobic transitions ; Escherichia coli
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie , Chemie und Pharmazie , Medizin
    Notizen: Abstract This study uncovers a new mechanism of regulation of DNA supercoiling operativein vivo upon an aerobic-anaerobic transition inEscherichia coli. Exponentially growing aerobic batch cultures were subjected to a shift to anaerobic conditions. The ratio [ATP]/[ADP] remained essentially constant at 8.5 in the aerobic culture and after a transition to anaerobiosis while DNA supercoiling increased noticeably upon anaerobiosis. This result indicated that the mechanism of regulation of DNA supercoiling by the [ATP]/[ADP] ratio was not operative. The increase in DNA supercoiling was followed by a large decrease in the DNA-relaxing activity of topoisomerase I while gyrase activity remained relatively constant. This decrease in the activity of topoisomerase I is likely to be responsible for the increase in DNA supercoiling.
    Materialart: Digitale Medien
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  • 5
    Digitale Medien
    Digitale Medien
    Springer
    Molecular and cellular biochemistry 85 (1989), S. 9-17 
    ISSN: 1573-4919
    Schlagwort(e): gangliosides ; ganglioside galactosaminyltransferase ; phospholipids and galactosaminyltransferase regulation
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie , Chemie und Pharmazie , Medizin
    Notizen: Summary A preparation of UDP-Ga1NAc:sialosyl lactosylceramide N-acetylgalactosaminyl transferase (E.C. 2.4.1.92) obtained from 14-day chick embryo brain was delipidated partially by treatment with cold acetone in the presence of varying amounts of sodium dodecyl sulphate (SDS). The lipid content and the enzyme activity of the preparation decreased as the concentration of SDS increased. At 0.3% SDS the lipid content was about 30% and the enzyme activity about 15% of the original. The activity could be restored up to 60% of the original by added phospholipids, provided the removal of endogenous lipids did not exceed 70%. Phospholipids with diferent composition showed different abilities to restore the enzyme activity. Among phosphatidylcholines the decreasing order of effectivity was dilauroyl → dimiristoyl → dipalmitoyl → distearoyl-choline. Dimiristoyl phosphatidylcholine, dimiristoyl phosphatidylglycerol and dipalmitoyl phosphatidylglycerol activated the enzyme more effectively than dimiristoyl phosphatidyl ethanolamine, dimiristoyl phosphatidic acid or brain phosphatidylserine. No correlation was found between the activating ability and the charge in the polar head group of the lipid added. Addition of dilauroyl phosphatidylcholine to the delipidated preparation increased about 5-fold the Vmax without affecting the apparent Km for both the donor nucleotide and acceptor glycolipid. The data suggest that the lipid composition of the enzyme environment constitutes a potential level of regulation of the activity of this key enzyme of ganglioside biosynthesis.
    Materialart: Digitale Medien
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  • 6
    Digitale Medien
    Digitale Medien
    Springer
    Molecular and cellular biochemistry 120 (1993), S. 1-13 
    ISSN: 1573-4919
    Schlagwort(e): dynamical organization ; levels of organization ; allometry ; bistability ; oscillations ; asymmetry changes
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie , Chemie und Pharmazie , Medizin
    Notizen: Summary Different levels of organization distinguished by characteristics spatial dimensions, Ec, and relaxation times, Tr, of biological processes ranging from electron transport in energy transduction to growth of microbial and plant cells, are shown to be related through a relation that may be interpreted as allometric and characterized by two different slopes. Processes, at levels of organization occurring in spatial dimensions of micrometers and relaxing in the order of minutes, delimit a ‘transition point’ between the two curves, that we interpret as a limit for the emergence of macroscopic coherence. The characteristic spatial dimension, Ec, and the relaxation time, Tr, contain dynamical information about the processes occurring at a given level of organization. When a steady state of a biological process at a certain level of organization becomes unstable, the system undergoes a transition to another level of organization. To exemplify the appearance of macroscopic order at levels of organization further from the ‘transition point’ we present in this report various experimental systems involving many levels of organization allometrically related that exhibit different kinds of self-organized behavior, i.e. bistability, oscillations, changes in (a)symmetry.
    Materialart: Digitale Medien
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  • 7
    ISSN: 1572-9699
    Schlagwort(e): Sporulation ; Saccharomyces cerevisiae ; 2 deoxy glucose ; metabolic fluxes ; gluconeogenesis ; glyoxylate cycle
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract Quantitative studies of metabolic fluxes during Saccharomyces cerevisiae sporulation on acetate in the presence of the glucose analog, 2-deoxy glucose (2dG) are reported. We have studied the inhibition of sporulation and associated catabolic or anabolic fluxes by 2dG. Sporulation frequencies decreased from 50% to 2% asci per cell at 2dG concentrations in the range of 0.03 to 0.30 g l〉-1, respectively. Under the same conditions, the acetate consumption flux was inhibited up to 60% and the glyoxylate cycle and gluconeogenic fluxes decreased from 0.7 and 0.3 mmol h〉-1 g〉-1 dw, respectively, to negligible values. We observed a linear correlation of the acetate consumption rate with the sporulation frequency by varying the 2dG concentration. The linear correlation was also verified between the frequency of sporulation and the fluxes through glyoxylate cycle and gluconeogenic pathways. In addition, the same association of inhibition of sporulation and metabolic fluxes was found in other S. cerevisiae strains displaying different potentials of sporulation. The results presented suggest that inhibition of sporulation in the presence of the glucose analog may be attributed, at least in part, to the inhibition of anabolic fluxes and might be associated with catabolite repression.
    Materialart: Digitale Medien
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  • 8
    Digitale Medien
    Digitale Medien
    Springer
    Antonie van Leeuwenhoek 69 (1996), S. 257-265 
    ISSN: 1572-9699
    Schlagwort(e): acetate and oxygen consumption ; Saccharomyces cerevisiae ; sporulation
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract We have quantified yeast carbon and oxygen consumption fluxes and estimated anabolic fluxes through glyoxylate and gluconeogenic pathways under various conditions of sporulation on acetate. The percentage of sporulation reached a maximum of 55% to 60% after 48 h in sporulation medium, for cells harvested from logarithmic growth in acetate minimal medium. When cells were harvested in the stationary phase of growth before transfer to sporulation medium, the maximum percentage of sporulation decreased to 40% along with the occurrence of meiosis as could be judged by counting of bi- and tetra-nucleated cells. In both experiments, the rates of acetate and oxygen consumption decreased as a function of time when exposed to sporulation medium. Apparently, the decrease of metabolic rates was not due to alkalinization. By systematically varying the cell concentration in sporulation medium from 1.4×107 to 20×107 cell ml-1, the percentage of sporulating cells was found to decrease in parallel with the rate of acetate consumption. When the sporulation efficiency attained under the different experimental conditions was plotted as a function of the rate of acetate consumption, a linear correlation was found. Anabolic fluxes estimation revealed a decrease of the rate through gluconeogenic and glyoxylate pathways occurring during sporulation progression. The pattern of metabolic fluxes progressively evolved toward a predominance of more oxidative catabolic fluxes than those exhibited under growth conditions. The results obtained are discussed in terms of a characteristic pattern of metabolic fluxes and energetics, associated to the development of yeast sporulation.
    Materialart: Digitale Medien
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  • 9
    ISSN: 1432-0991
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie , Medizin
    Notizen: Abstract. We have studied the effect of disrupting catabolite (de)repression genes SNF1, SNF4, and MIG1 on the cell cycle behavior of the CEN.PK122 wild type (WT) strain of Saccharomyces cerevisiae by flow cytometry in glucose-limited chemostat cultures or batch growth in the presence of different carbon sources. Through a combination of flow cytometry of propidium iodide–stained cells and mathematical modeling we showed that the deletion of the SNF4 gene provoked a decrease in the length of G1 with respect to the WT strain along with a smaller difference in the cell cycle length of parent and daughter cells. snf1 and mig1 mutants exhibited slightly shorter G1 respect to the WT. Additionally, in the mig1 mutant the cell cycle length of parent and daughter cells was slightly altered. The results obtained are in agreement with the view that the SNF4 gene is involved in the regulation of cell cycle in yeast.
    Materialart: Digitale Medien
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  • 10
    Digitale Medien
    Digitale Medien
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 37 (1991), S. 197-204 
    ISSN: 0006-3592
    Schlagwort(e): Chemistry ; Biochemistry and Biotechnology
    Quelle: Wiley InterScience Backfile Collection 1832-2000
    Thema: Biologie , Werkstoffwissenschaften, Fertigungsverfahren, Fertigung
    Notizen: Thermodynamic and kinetic calculations were performed in a model of mixed substrate metabolism. The model simulates the catabolic breakdown of a first substrate, glucose (S1), in the presence of a second substrate, formate (S2), which acts as an additional source of free energy. The principal results obtained with different relative rates of uptake of S2 allow to predict and interpret the following experimental observations: (1) the existence of increased ATP yields by mixed substrate utilization with a maximum ATP yield and optimum input (or molar) ratio for both substrates; (2) a greater assimilation of S1 which may be interpreted as a decreasing fraction of energy required for assimilation; (3) a decrease in ATP yields due to increasing energy demand for transport; (4) an increased assimilation of the carbon source (S1) as a function of increasing inputs of the additional energy source; (5) thermodynamic efficiency (η) defined as the ratio between the output power of ATP synthesis and the input catabolic power, increases for S2/S1 ratios ranging between 0.08 and 2 while for ratios higher than two a slight decrease of η was noticed; and (6) the observed maximum in ATP yield for optimum molar ratio of the two substrates corresponds to high η predicting that higher biomass yields may be obtained through a variable, high, η by chanelling fluxes through catabolic pathways with different ATP yields. During oscillatory behavior, maxima in fluxes were almost coincident with maxima in forces. Thus, the pattern of dissipation was not so advantageous as in the single substrate model under starvation conditions.
    Zusätzliches Material: 10 Ill.
    Materialart: Digitale Medien
    Bibliothek Standort Signatur Band/Heft/Jahr Verfügbarkeit
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