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  • 1
    Electronic Resource
    Electronic Resource
    Berlin : Wiley-Blackwell
    Acta Biotechnologica 16 (1996), S. 271-282 
    ISSN: 0138-4988
    Keywords: Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Process Engineering, Biotechnology, Nutrition Technology
    Notes: A mathematical model has been developed to describe the stability behaviour of the pBR322 plasmid derivative pBB210 with the β-lactamase gene and the human interferon-α1 gene in Escherichia coli TG1 under non-selective, selective and modified selective conditions in a chemostat. The model was formulated on the basis of experimental investigations. It includes the interaction between β-lactam antibiotics (ampicillin and sulbactam) and cells (with and without plasmids), in particular the correlation between the growth rate of plasmid-free cells and ampicillin concentration in the medium; ampicillin transport into the periplasm of the plasmid-bearing cells; ampicillin degradation in the periplasm by by plasmid-encoded β-lactamase and the inhibition of the latter by sulbactam.The results obtained by the simulation of chemostat cultivations under various conditions and by steady state analyses are closely related to the results of experiments. Under non-selective conditions, the fraction of plasmid-bearing cells was approaching zero. Under selective and modified selective conditions, a coexistence between plasmid-free and plasmid-bearing cells was reached at steady state. Under these conditions, the steady state fraction of plasmid-bearing cells was proportional to the ampicillin concentration in the feed and inversely proportional to the cell concentration in the chemostat. During high-density cultivation, a large amount of ampicillin is necessary to suppress plasmid-free cells. Even small concentrations of the β-lactamase inhibitor sulbactam in the feed increased the steady state fraction of plasmid-bearing cells (from 17.2% to 99.6% at sulbactam-Na concentrations of 0 to 5 mg/l).
    Additional Material: 5 Ill.
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  • 2
    ISSN: 0138-4988
    Keywords: Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Process Engineering, Biotechnology, Nutrition Technology
    Notes: No Abstracts.
    Type of Medium: Electronic Resource
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  • 3
    Electronic Resource
    Electronic Resource
    Berlin : Wiley-Blackwell
    Acta Biotechnologica 16 (1996), S. 303-314 
    ISSN: 0138-4988
    Keywords: Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Process Engineering, Biotechnology, Nutrition Technology
    Notes: Pilot plant studies were performed using a concentric-tube airlift bioreactor of 2.5 m3 fermentation volume. The results have proven the relative merits of such a system in the biosynthesis of nystatin, produced by Streptomyces noursei, in submerged aerobic cultivation and batch operation mode. The results were compared to those obtained in a pilot-scale stirred tank bioreactor of 3.5 m3 fermentation volume.The fermentation processes in the two fermentation devices were similar with respect to substrate utilization, biomass production and nystatin biosynthesis.In the riser section, the dissolved oxygen concentration was higher than that in the downcomer.The volumetric oxygen mass transfer coefficient was dependent on the rheological behaviour of the biosynthesis liquids, which was not constant during the fermentation process.The total energy consumption for nystatin production in the airlift bioreactor was 56% of that in the stirred tank, while the operating costs represented 78% of those in the stirred tank bioreactor.
    Additional Material: 6 Ill.
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  • 4
    Electronic Resource
    Electronic Resource
    Berlin : Wiley-Blackwell
    Acta Biotechnologica 16 (1996) 
    ISSN: 0138-4988
    Keywords: Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Process Engineering, Biotechnology, Nutrition Technology
    Type of Medium: Electronic Resource
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  • 5
    Electronic Resource
    Electronic Resource
    Berlin : Wiley-Blackwell
    Acta Biotechnologica 16 (1996), S. 321-327 
    ISSN: 0138-4988
    Keywords: Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Process Engineering, Biotechnology, Nutrition Technology
    Notes: This study focuses on the growth of Zymomonas mobilis strain 113 S and its ethanol and levan production under the conditions of increasing sucrose medium osmolality caused by NaCl, KCl, sorbitol or maltose. The increase in medium osmolality (700-1,500 mosml/kg) was accompanied by the inhibition of growth (growth rate, biomass yield) and ethanol production (specific productivity and yield) In contrast, levan synthesis was less affected or even stimulated and, as a consequence, levan specific productivity was increased significantly. A decrease in the anabolic growth parameters correlated with a parallel inhibition of glucose-6-P dehydrogenase and alcohol dehydrogenase (isoenzyme ADH II) activities. A significant inverse linear relationship (r = - 0.932, 1 - P = 0.01) was observed between the values of the specific productivities of ethanol and levan. This relationship was confirmed independently by a controlled reduction of growth and ethanol productivity (3.75-4.75 mM sodiumbisulphite as an acceptor of acetaldehyde formed in the pyruvate decarboxylase reaction). As further support of this relationship, a significant inverse correlation was observed between levan specific productivity and ATP concentration in Zymomonas mobilis cells, most probably demonstrating that a reduced level of energetic metabolism is favourable for levan production.
    Additional Material: 7 Ill.
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  • 6
    Electronic Resource
    Electronic Resource
    Berlin : Wiley-Blackwell
    Acta Biotechnologica 16 (1996), S. 103-119 
    ISSN: 0138-4988
    Keywords: Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Process Engineering, Biotechnology, Nutrition Technology
    Notes: All the oceans are plentiful with marine algae. Non-viable marine macroalgae are able to adsorb heavy metal ions. Compared with other biosorbents, such as fungi, bacteria, yeasts and microalgae, they have the advantage of being easily available, cheap and having high heavy metal sorption capacities. The by-products of marine phaeophyceae are even more cost-effective heavy metal biosorbers.Experiments of heavy metal sorption using non-viable Fucus vesiculosus, Ascophyllum nodosum and algal by-products were carried out to investigate the factors influencing and optimizing the heavy metal biosorption.The pH value, biomass concentration, heavy metal concentration, heavy metal species, competing ions, algal varieties and time were the most decisive parameters. The sorption isotherms showed increasing sorption capacities and decreasing sorption efficiencies with an increase in the initial heavy metal concentration. Sorption kinetics of different metals were established. Biomass concentration influenced the sorption efficiencies very much, but reduced the sorption capacity per g biomass. The pH value controlled the sorption (pH 3-7) and desorption (pH 1-2) decisively.Beside heavy metal contaminated model waters, actual industrial effluents were treated successfully by algal sorbents in batch experiments and continuous column tests.Transmission electron micrographs of different contaminated and untreated algal specimens are available.
    Additional Material: 10 Ill.
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  • 7
    ISSN: 0138-4988
    Keywords: Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Process Engineering, Biotechnology, Nutrition Technology
    Notes: A bacterial strain, called P4a, was isolated from debris of concrete samples of a demolished herbicide factory. The samples were contaminated with chlorinated and methylated phenoxyalkanoic acids. Strain P4a was able to utilize 2,4-dichloro- (2,4-D) and 4-chloro-2-methylphenoxyacetic acid (MCPA) as the sole source of carbon and energy; degradation of 2,4-dichlorinated and 4-chloro-2-methylated phenoxypropionic acid and -butyric acid derivatives was not found. Growth on 2,4-D was observed from a pH of 5.6 up to a pH of about 10, with optimum growth at around 8.5. No supplements were found to be required for growth on 2,4-D, but the presence of yeast extract increased the growth rate from less than 0.05 h-1 to 0.2 h-1. The strain was metabolically active up to pH values of 12, which corresponded to the pH of aqueous eluates from such material. It was able to degrade 2,4-D under these conditions up to an initial concentration of 400 mg/1 and in fact did degrade 1,600 mg/l of 2,4-D at an initial pH value of 11. Strain P4a was tentatively identified as Comamonas acidovorans on the basis of the substrate utilization pattern (BIOLOG), fatty acid profile (MIS) and G+C content.
    Additional Material: 4 Ill.
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  • 8
    Electronic Resource
    Electronic Resource
    Berlin : Wiley-Blackwell
    Acta Biotechnologica 16 (1996), S. 133-144 
    ISSN: 0138-4988
    Keywords: Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Process Engineering, Biotechnology, Nutrition Technology
    Notes: Xylitol production from hardwood hemicellulosic hydrolysates by well-known ethanol-producing yeasts was stimulated through an experimental schedule including pretreatments of the hydrolysate, the choice of the best xylitol producer and the selection of the optimum fermentation conditions. The xylitol or ethanol yields obtained on consumed xylose demonstrated that their production was stimulated under completely different conditions, as to be expected by the fact that these catabolites are the final products of different metabolic pathways. In particular, the catabolism of Pachysolen tannophilus, that is the best ethanol producer from this natural substrate, could be targeted towards xylitol rather than towards ethanol production by ensuring a strongly reducing environment through a suitable pretreatment of the hydrolysate. The final removal of fermentation inhibitors by adsorption onto highly adsorbing substances allowed a further 20% xylitol yield increase.
    Additional Material: 6 Ill.
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  • 9
    Electronic Resource
    Electronic Resource
    Berlin : Wiley-Blackwell
    Acta Biotechnologica 16 (1996), S. 79-79 
    ISSN: 0138-4988
    Keywords: Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Process Engineering, Biotechnology, Nutrition Technology
    Notes: No Abstracts.
    Type of Medium: Electronic Resource
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  • 10
    ISSN: 0138-4988
    Keywords: Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Process Engineering, Biotechnology, Nutrition Technology
    Notes: The xylose conversion into by Candida guilliermondii was evaluated in sugar cane bagasse hemicellulosic hydrolysate. The effect of air flow rates of 0.4, 0.6 and 0.8 vvm cn xylitol formation was studied. In addition, inoculum previously adapted to the hydrolysate was also tested in the fermentation carried out at 0.6 vvm. The results showed that xylitol production depends markedly on the aeration rate and on the previous adaptation of the yeast to the hydrolysate. When the highest productivity of xylitol was 0.39 g/l × h. However, during the fermentation carried out at an air flow rate of 0.6 vvm with adapted inoculum, the productivity increased to 0.65 g/l × h. Furthermore, the adapted cells performed quite well in the presencel of acetic concentrations of about 4.5 g/l in the medium.
    Additional Material: 2 Ill.
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  • 11
    Electronic Resource
    Electronic Resource
    Berlin : Wiley-Blackwell
    Acta Biotechnologica 16 (1996), S. 81-88 
    ISSN: 0138-4988
    Keywords: Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Process Engineering, Biotechnology, Nutrition Technology
    Notes: Glucose oxidase and peroxidase were immobilized individually to a urea derivative of granulated microcrystallized cellulose activated by formaldehyde. The catalytic properties of the immobilized enzymes were studied and compared to those of the soluble enzymes. The immobilized glucose oxidase and peroxidase were used for the manual determination of the concentration of glucose in sera. The developed method is characterized by high analytical reliability and comparatively low cost. The results correlated well with those obtained by using a BECKMAN glucoanalyzer, utilizing soluble glucose oxidase.
    Additional Material: 4 Ill.
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  • 12
    Electronic Resource
    Electronic Resource
    Berlin : Wiley-Blackwell
    Acta Biotechnologica 16 (1996), S. 247-255 
    ISSN: 0138-4988
    Keywords: Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Process Engineering, Biotechnology, Nutrition Technology
    Notes: Waste water, derived from the reprocessing of used emulsions or suspensions, contains high concentrations of emulsified mineral oil and stabilizers, as well as different additives that are needed during the treatment process.Two stirred-tank reactors and two fixed-bed reactors were used to study the biodegradation of these waste-water compounds during two-stage biological treatment. The waste water was first proceesed in an activated sludge reactor to remove easily biodegradable substances. The effluent from the first stage was treated in three parallel operating reactors: an activated sludge tank containing different amounts of powdered activated carbon (PAC, between 0 and 2%), an upflow anaerobic fixed-bed reactor and an aerobic fixed-bed reactor (trickling filter). The results from the continuous treatment were compared with laboratory batch experiments.About 60% of the influent TOC was reduced by the first activated sludge treatment. The removal efficiency increased to about 70% by using a second activated sludge stage. This degradation was comparable to the maximum degree of degradation measured in laboratory batch experiments. PAC addition to the second activated sludge tank resulted in increased degradation rates. The removal efficiency increased to about 76% when 0.1% PAC was added and to 96% with 1% PAC. The removal efficiency decreased to 84% when the proportion of PAC was further increased to 2%. Variations in the amount of PAC addition per unit influent volume in the range of 50 and 200 mg/l had no significant effect on the TOC removal.Degradation models based on the MONOD-type equation were found to be in close correlation with the results obtained from batch experiments. However, the biological removal rates measured in batch experiments did not reflect the removal capacity determined in continuous operating treatment systems.
    Additional Material: 5 Ill.
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  • 13
    Electronic Resource
    Electronic Resource
    Berlin : Wiley-Blackwell
    Acta Biotechnologica 16 (1996), S. 283-292 
    ISSN: 0138-4988
    Keywords: Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Process Engineering, Biotechnology, Nutrition Technology
    Notes: The growth of a thermophilic Clostridium sp. and the production of α-glucosidase, α-amylase and pullulanase were studied under anaerobic conditions using different carbon and nitrogen sources and varying pH values and temperatures. Growth and enzyme activities were highest with soybean meal as the nitrogen source. The optimum concentration was 2.5% [w/v] for the production of α-amylase as well as pullulanase and 2% [w/v] for α-glucosidase. The best carbon source proved to be soluble starch for α-amylase, and pullulanase and maltose for α-glucosidase. Growth and enzyme production reached their optimum at pH 6.5 to 7.0 and 70°C. Under these conditions, the enzyme activities followed exponential growth with maximum yields of α-glucosidase, α-amylase and pullulanase at 28, 36, and 44 h.
    Additional Material: 5 Ill.
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  • 14
    ISSN: 0138-4988
    Keywords: Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Process Engineering, Biotechnology, Nutrition Technology
    Notes: Instead of yeast extracts, casein hydrolysates from several suppliers were added to culture media and analyzed with respect to their ability to stimulate β-galactosidase activity in Kluyveromyces bulgaricus cells. Four enzymatic casein hydrolysates caused a significantly higher stimulation of enzyme activity, while acid casein hydrolysates clearly reduced the enzyme activity. Enzymatic casein hydrolysates, inducing high and low lactase activity, were analyzed with respect to average peptide length (APL), vitamins (niacin, pathothenate) as well as free and bound amino acids. The molecular weights of these casein hydrolysates were estimated by gel filtration. No correlation was found between the degree of enzyme stimulation and the vitamin contents, the APL values and the free amino acid contents of the casein hydrolysates. Casein hydrolysates stimulating the lactase activity were less soluble in water and, in a gel filtration column, they showed three peaks with slightly lower molecular weights than the three peaks seen in hydrolysates which had no effect on activity. APL values of alcoholic precipitates of casein hydrolysates showed an inverse correlation to lactase activity. The molecular weights of alcoholic precipitates of lactase stimulating digests were also lower compared to non-stimulating ones. Alcoholic precipitates with lactase-stimulating activity were more hydrophilic, as was shown by a smaller proportion absorbed in a C18 column and by amino acid analysis. Our results sugest that alcoholic precipitates could probably be important in the lactase stimulation and their composition should be further investigated. The mechanism is nevertheless complex and may be caused by various simultaneous factors.
    Additional Material: 4 Ill.
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  • 15
    Electronic Resource
    Electronic Resource
    Berlin : Wiley-Blackwell
    Acta Biotechnologica 16 (1996), S. 145-153 
    ISSN: 0138-4988
    Keywords: Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Process Engineering, Biotechnology, Nutrition Technology
    Notes: The influence of static mixers on mixing time and circulation time in an external-loop airlift bioreactor with gas-induced and forced-liquid circulation was investigated.The study was carried out with water and three viscous, non-Newtonian starch solutions. The mixing time was determined for the overrall flow loop using a classical tracer response technique.It was found that the mixing time is highly dependent on the superficial gas velocity and the presence of static mixers. The shortest mixing time is achieved in a forced-loop airlift reactor without static mixers, where the average mixing time value is only 1/3 of the time necessary for mixing in the airlift reactor with gas-induced liquid circulation and static mixers.The pseudoplasticity of the liquid phase insignificantly influences the mixing time and the circulation time.
    Additional Material: 3 Ill.
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  • 16
    Electronic Resource
    Electronic Resource
    Berlin : Wiley-Blackwell
    Acta Biotechnologica 16 (1996), S. 155-162 
    ISSN: 0138-4988
    Keywords: Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Process Engineering, Biotechnology, Nutrition Technology
    Notes: The Rhizobium sp., isolated from the root nodules of the leguminous climbing herb, Tephrosia Purpurea Pers., produced high amounts of extracellular polysaccharides (EPS) in yeast extract mannitol medium. Growth and EPS production started simultaneously, but the production reached its maximum in the stationary growth phase of the bacteria. Attempts were made to optimize the cultural requirements for growth and maximum EPS production. The EPS production was increased by 72.5% over the control when the medium was supplemented with mannitol (2%), thiamine hydrochloride (5 μg/ml) and KNO3 (0.1%). The EPS contained glucose, galactose and mannose monomers. The possible role of the rhizobial EPS was discussed.
    Additional Material: 1 Ill.
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  • 17
    ISSN: 0138-4988
    Keywords: Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Process Engineering, Biotechnology, Nutrition Technology
    Notes: No Abstracts.
    Type of Medium: Electronic Resource
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  • 18
    ISSN: 0138-4988
    Keywords: Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Process Engineering, Biotechnology, Nutrition Technology
    Notes: No Abstracts.
    Type of Medium: Electronic Resource
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  • 19
    ISSN: 0138-4988
    Keywords: Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Process Engineering, Biotechnology, Nutrition Technology
    Notes: No Abstracts.
    Type of Medium: Electronic Resource
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  • 20
    ISSN: 0138-4988
    Keywords: Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Process Engineering, Biotechnology, Nutrition Technology
    Notes: Extracellular acid phosphatase was studied as a minor enzyme of the fungal strain Humicola lutea 120-5 having a clear relation to the secretion of acid proteinases. A medium lacking in mineral orthophosphates ensured a fivefold higher yield of phosphatase while the proteinase production was reduced. An acid phosphatase fraction free of proteinase activity was isolated demonstrating a maximum hydrolysis of 4-nitrophenyl-phosphate at a pH of 4.0 and 50°C. The phosphatase catalyzed a partial dephosphorylation of up to 30% of casein at a pH of 3.0 causing a complete substrate precipitation. Both proteinase and phosphatase biosynthesis increased twofold when natural casein was replaced by partially dephosphorylated casein in the cultivation medium.
    Additional Material: 3 Ill.
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  • 21
    Electronic Resource
    Electronic Resource
    Berlin : Wiley-Blackwell
    Acta Biotechnologica 16 (1996), S. 245-246 
    ISSN: 0138-4988
    Keywords: Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Process Engineering, Biotechnology, Nutrition Technology
    Notes: No Abstracts.
    Type of Medium: Electronic Resource
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  • 22
    Electronic Resource
    Electronic Resource
    Berlin : Wiley-Blackwell
    Acta Biotechnologica 16 (1996), S. 237-244 
    ISSN: 0138-4988
    Keywords: Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Process Engineering, Biotechnology, Nutrition Technology
    Notes: The remediation of polluted soils by microbial communities is a complex process, characterized by many competitive and cooperative mutual relations between the individual organisms. For the overall characteristic of such processes, top-down systems analyses promise a better description for prognostic aims than bottom-up approaches.This is demonstrated using the EVOLON model to fit the cumulative O2 consumption data of a microbial ecosystem metabolizing diesel fuel in polluted soil. The EVOLON is a generic top-down model providing close approximation to the experimental data.The advantage of using this model instead of other similar models is demonstrated by a comparison of the deviations between model values and experimental data (residuals).
    Additional Material: 4 Ill.
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  • 23
    ISSN: 0138-4988
    Keywords: Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Process Engineering, Biotechnology, Nutrition Technology
    Notes: No Abstracts.
    Type of Medium: Electronic Resource
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  • 24
    ISSN: 0138-4988
    Keywords: Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Process Engineering, Biotechnology, Nutrition Technology
    Notes: No Abstracts.
    Type of Medium: Electronic Resource
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  • 25
    Electronic Resource
    Electronic Resource
    Berlin : Wiley-Blackwell
    Acta Biotechnologica 16 (1996), S. 120-120 
    ISSN: 0138-4988
    Keywords: Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Process Engineering, Biotechnology, Nutrition Technology
    Notes: No Abstracts.
    Type of Medium: Electronic Resource
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  • 26
    ISSN: 0138-4988
    Keywords: Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Process Engineering, Biotechnology, Nutrition Technology
    Notes: Twelve bacterial isolates, four of them assigned to the genus Acinetobacter, were taken from sewage of a treatment plant with Enhanced Biological Phosphorus Removal (EBPR) and screened for phosphorus uptake, polyphosphate (polyP) accumulation and adsorption under limited carbon and nitrogen conditions. In addition, poly-β-hydroxyalkanoate (PHA) production was studied under carbon, nitrogen, phosphorus, and oxygen limitation. Under C limitation, the uptake of phosphorus was highest, ranging up to 66 mg P per g dry weight (dw) for the Acinetobacter isolates, whereas the highest amount of polyP was detected under limited N conditions (up to 25 mg PolyP / g dw). Extra-cellular polyP was detected, however to a minor extent, accounting for a maximum of 10% of the total polyP in one Acinetobacter isolate. The highest PHA concentration (given as 3-hydroxybutyrate, 3 HB) with 211 mg 3 HB / g dry weight (21% of the dried cell mass) was found for A. johnsonii 120 under nitrogen limitation, but also under P and O2 limitation, PHA, mainly poly-β-hydroxybutyrate and poly-β-hydroxyvalerate, were produced. Three isolates, assigned to the genus Pseudomonas, showed even higher values (345-427 mg 3 HB / g dw) under N limitation. Studies with Acinetobacter johnsonii 120 in continuous culture, simulating the aerobic/anaerobic periods of a waste-water treatment plant, resulted in a P elimination of 36% at an anaerobic contact time of 0.6 h. This value increased to 51% at an anaerobic contact time of 3.1 h. No release of phosphate and no uptake of acetate could be detected during the anaerobic period. In addition, Acinetobacter johnsonii 120 was not able to synthesize PHA under anaerobic conditions. By changing the anaerobic conditions to aerobic, a continuous decrease of the polyP content relative to the totalP content from 45% (day 1 of the aerobic process) to 19% (day 17 of the aerobic process) was observed. The amount of PHA increased to 50.4 mg 3 HB/g dw under aerobic conditions. The results indicate again that the EPBR process cannot be defined by simply applying the knowledge of the metabolic processes, observed or assumed in Acinetobacter pure cultures, to the complexity of the process in sewage treatment plants.
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  • 27
    Electronic Resource
    Electronic Resource
    Berlin : Wiley-Blackwell
    Acta Biotechnologica 16 (1996), S. 132-132 
    ISSN: 0138-4988
    Keywords: Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Process Engineering, Biotechnology, Nutrition Technology
    Notes: No Abstracts.
    Type of Medium: Electronic Resource
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  • 28
    Electronic Resource
    Electronic Resource
    Berlin : Wiley-Blackwell
    Acta Biotechnologica 16 (1996), S. 19-30 
    ISSN: 0138-4988
    Keywords: Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Process Engineering, Biotechnology, Nutrition Technology
    Notes: The fundamentals of the biological treatment of contaminated soils were investigated in bioreactors with the aim to optimize the processes of biological soil treatment in order to achieve the highest possible degree of degradation within the shortest period of time. Preinvestigations using test systems at different scales have provided information about the possibilities of enhancing the decomposition processes which are dependent on various factors, such as milieu conditions, additives, etc., that must be known before remedial actions are taken.The investigations made so far have shown that compost is a favourable additive when oil-contaminated soils are biologically treated. The degradation of contaminants can be enhanced by the addition of compost. This positive effect is attributed to various mechanisms. In this paper, results of a variety of test systems at different scales are presented.In test series, different amounts of biocompost were added to investigate the influence on the degradation of diesel fuel. It was demonstrated that by increasing the compost content - the cumulative O2 consumption caused by the degradation of the diesel fuel contaminants increased.It could be shown that the reduction of the diesel fuel contaminants in the soil was independent of the compost age and amounted to approximately 94% of the initial quantity.The addition of biocompost could also enhance the degradation of real contaminants. After a test period of 162 days in set-ups with compost addition, more than 75% of the lubricating oil contaminants disappeared, while less than 37% of the contaminants disappeared in set-ups without compost addition.Moreover, by the addition of compost, the formation of pellets during the dynamic treatment of soil materials could be reduced.
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  • 29
    Electronic Resource
    Electronic Resource
    Berlin : Wiley-Blackwell
    Acta Biotechnologica 16 (1996), S. 43-56 
    ISSN: 0138-4988
    Keywords: Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Process Engineering, Biotechnology, Nutrition Technology
    Notes: The data presented here with respect to the behaviour of industrial scale stirred tank bioreactors equipped with modified RUSHTON turbine agitators in the biosynthesis processes of antibiotics are valid for that case that the power consumption is the same as it is in standard RUSHTON turbine agitators.Each modified RUSHTON turbine agitator was obtained through the variation of the blade surface by adding perforations so that the ratio between the perforation surface area and the full surface area (or the surface fraction of the perforations) is 0.36.In the fermentations of Streptomyces aureofaciens, Streptomyces rimosus and Penicillium chrysogenum producing tetracycline, oxytetracyline and penicillin, respectively, in bioreactors equipped with modified RUSHTON turbine agitators, the relative antibiotic production is increased by more than 30% compared to standard bioreactors.
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  • 30
    Electronic Resource
    Electronic Resource
    Berlin : Wiley-Blackwell
    Acta Biotechnologica 16 (1996), S. 57-64 
    ISSN: 0138-4988
    Keywords: Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Process Engineering, Biotechnology, Nutrition Technology
    Notes: Bacillus larvae, the causative agent of American foulbrood in honey bees completes its life cycle of germination, outgrowth and sporulation in young honey bee larvae by killing them and often bringing about the destruction of the entire hive. While B. larvae germinates and outgrows on complex organic media in vitro, the literature suggests, for reasons that are not at all clear, that a relatively large number of spores of B. larvae are required to yield each visible colony (colony forming units, CFU) on media. Various researchers have reported that from 16 to 3,000 or more spores of B. larvae are required to yield a single colony on an agar plate. HANSEN in Denmark designed a useful method of spreading approximately 80 mg of honey directly on the surface of a PETRI plate containing “J” agar medium to determine if B. larvae spores are present in the honey. In the present study, selected media were tested for the ability to recover B. larvae spores in honeys in the form of visible colonies (CFU) using HANSEN's strek method. A modification of a medium (TMYGP) developed by DINGMAN and STAHLY, (T-HCL-YGP agar), recovered considerably more viable B. larvae spores in the form of visible colonies (CFU) than HANSEN's “J” medium. When “J” medium was fortified with 0.1% sodium pyruvate, it was comparable to modified T-HCL-YGP medium in its recovery of B. larvae spores. Brain heart infusion agar (BHIA) with the addition of thiamine recovered more spores in the form of viable colonies than did “J” medium but it was not as efficient as T-HCL-YGP medium. Serial dilution from 100 to 10,000 times of weighed samples of honey with deionized water led to higher spore counts (CFU per g honey) than that indicated by undiluted honeys plated at 80 mg levels directly onto the surface of media by the HANSEN procedure.
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  • 31
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    Berlin : Wiley-Blackwell
    Acta Biotechnologica 16 (1996), S. 80-80 
    ISSN: 0138-4988
    Keywords: Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Process Engineering, Biotechnology, Nutrition Technology
    Notes: No Abstracts.
    Type of Medium: Electronic Resource
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  • 32
    Electronic Resource
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    Berlin : Wiley-Blackwell
    Acta Biotechnologica 16 (1996) 
    ISSN: 0138-4988
    Keywords: Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Process Engineering, Biotechnology, Nutrition Technology
    Type of Medium: Electronic Resource
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  • 33
    ISSN: 0138-4988
    Keywords: Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Process Engineering, Biotechnology, Nutrition Technology
    Notes: No Abstracts.
    Type of Medium: Electronic Resource
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  • 34
    ISSN: 0138-4988
    Keywords: Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Process Engineering, Biotechnology, Nutrition Technology
    Notes: No Abstracts.
    Type of Medium: Electronic Resource
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  • 35
    Electronic Resource
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    Berlin : Wiley-Blackwell
    Acta Biotechnologica 16 (1996), S. 257-270 
    ISSN: 0138-4988
    Keywords: Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Process Engineering, Biotechnology, Nutrition Technology
    Notes: In previous studies, the white-rot basidiomycete Lentinus edodes, strain SC-495, was proved to be a “selective” lignin degrader and its extracellular crude preparations arising from solid-state cultures were successfully employed in biopulping experiments on annual plants. This fungus produced extracellular laccase as the predominant phenoloxidase when growing in solid-state fermentation on corn stalks. Laccase from this strain was purified and partially characterized, as an initial approach towards the study of its ligninolytic complex. Laccase was purified 69.6-fold by anion-exchange chromatography and two affinity-chromatography steps with an overall yield of 7.45%. The native enzyme exhibited a molecular mass of 74 kDa, an isoelectric point of 3.42 and a carbohydrate content of 7.5%. The absorption spectrum of laccase showed a maximum at 605 nm, typical of blue-copper oxidases. The optimum pH and temperature for the activity of laccase were 4.0-4.2 and 50°C, respectively. Kinetic experiments, performed with a wide range of phenolic compounds, showed that the reaction rate and the substrate affinity greatly varied depending on the nature of substituents and their reciprocal positions on the aromatic ring. In particular, the enzyme showed high affinity to phenolic compounds bearing methoxyl or methyl groups, but no affinity to those bearing the nitro group directly attached to the benzene ring, nor to non-phenolic lignin-related compounds, such as trans-cinnamic acid or 3,4-dimethoxycinnamic acid. The huge differences in terms of reactivity of the enzyme towards phenolic compounds suggests that a preliminary systematic screening should be advisable when using laccase in effluent treatment applications.
    Additional Material: 5 Ill.
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  • 36
    ISSN: 0138-4988
    Keywords: Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Process Engineering, Biotechnology, Nutrition Technology
    Notes: No Abstracts.
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  • 37
    ISSN: 0138-4988
    Keywords: Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Process Engineering, Biotechnology, Nutrition Technology
    Notes: No Abstracts.
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  • 38
    ISSN: 0138-4988
    Keywords: Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Process Engineering, Biotechnology, Nutrition Technology
    Notes: No Abstracts.
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  • 39
    ISSN: 0138-4988
    Keywords: Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Process Engineering, Biotechnology, Nutrition Technology
    Notes: No Abstracts.
    Type of Medium: Electronic Resource
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  • 40
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    Berlin : Wiley-Blackwell
    Acta Biotechnologica 16 (1996), S. 193-198 
    ISSN: 0138-4988
    Keywords: Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Process Engineering, Biotechnology, Nutrition Technology
    Notes: The conditions for an optimum formation of protoplasts from mycelium of Pencillium citrinum, which produces extracellular lipase, were examined. The best results were achieved using Novozyme 234 in combination with β-glucuronidase. Chitinase or β-glucuronidase alone were not able to produce protoplasts under the given conditions. For the regeneration frequency, the type of osmotic stabilizer was important. The highest regeneration frequency (84.6% protoplasts) was achieved by using 0.6 M KCl as an osmotic stabilizer. Some regenerants were more effective in producing lipase than the parent. In particular, strain no. P-8 showed a three times higher lipolytic activity than the parent strain, but this high enzyme activity was unstable after subculture.
    Additional Material: 3 Tab.
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  • 41
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    Berlin : Wiley-Blackwell
    Acta Biotechnologica 16 (1996), S. 199-205 
    ISSN: 0138-4988
    Keywords: Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Process Engineering, Biotechnology, Nutrition Technology
    Notes: Tea fungus/kombucha, an acetic acid flavoured fermented tea beverage, is widely consumed in various parts of the world and has more recently become a fad in the United States. This is due in part to the fact that it can be produced in the home, and it is reported to be medicinal, effective against arthritis, psoriasis, chronic fatigue, constipation, indigestion and metabolic diseases. Among 264 references from 1852 to 1961, there are reports of antibiotic activity against Agrobacterium tumefaciens and medicinal value against a variety of diseases. The medicinal value appears to be related to that attributed to vinegar, one of our most ancient foods. We decided to test tea fungus/kombucha for its antibiotic activity against Helicobacter pylori, a primary cause of gastritis related to peptic ulcers and gastric carcinoma, Escherichia coli, Staphylococcus (Micrococcus) aureus and Agrobacterium tumefaciens. Tea containing 4.36 g of dry tea per litre and 10% of sucrose and fermented with the tea fungus showed no antibiotic activity in the beverage beyond that caused by acetic acid, a primary product of the fermentation.
    Additional Material: 2 Tab.
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  • 42
    ISSN: 0138-4988
    Keywords: Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Process Engineering, Biotechnology, Nutrition Technology
    Notes: No Abstracts.
    Type of Medium: Electronic Resource
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  • 43
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    Berlin : Wiley-Blackwell
    Acta Biotechnologica 16 (1996), S. 215-223 
    ISSN: 0138-4988
    Keywords: Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Process Engineering, Biotechnology, Nutrition Technology
    Notes: An investigation was carried out on the growth and ethanol production of a novel thermotolerant ethanol-producing Kluyveromyces marxianus IMB3 yeast strain. It grew aerobically on glucose, lactose, cellobiose, xylose and whey permeate and fermentad all the above carbon sources to ethanol at 45°C. This strain was capable of growing under anaerobic chemostat fermentation conditions at 45°C and a dilution rate of 0.15 h-1 and produced ≤ 0.9 g/l biomass and 1.8% (v/v) ethanol. An increase in biomass (up to 10.0 g/l) and ethanol (up to 4.3% v/v at 45°C and 7.7% v/v at 40°C) were achieved by applying a continuous two-stage fermentation in sequence (one aerobic and one anaerobic stage) or a two-stage anaerobic fermentation with cell recycling. Potential applications, involving alcohol production systems, for use in dairy and wood related industries, were discussed.
    Additional Material: 2 Ill.
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  • 44
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    Berlin : Wiley-Blackwell
    Acta Biotechnologica 16 (1996), S. 227-235 
    ISSN: 0138-4988
    Keywords: Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Process Engineering, Biotechnology, Nutrition Technology
    Notes: Viable microalgae are known to be able to accumulate heavy metals (bioaccumulation). Against a background of the increasing environmental risks caused by heavy metals, the microalgae Chlorella vulgaris and Spirulina platensis and their potential for the biological removal of heavy metals from aqueous solutions were taken as an example for investigation. Small-scale cultivation tests (50 1) with Cd-resistant cells of Chlorella vulgaris have shown that approx. 40% of the added 10 mg Cd/l was removed from the solution within seven days. At this heavy metal concentration sensitive cells died.Non-viable microalgae are able to eliminate heavy metal ions in a short time by biosorption in uncomplicated systems, without any toxicity problems. Compared with original biomasses, the sorption capacity of microalgal by-products changes only insignificantly. Their low price makes them economical.
    Additional Material: 2 Ill.
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  • 45
    ISSN: 0138-4988
    Keywords: Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Process Engineering, Biotechnology, Nutrition Technology
    Notes: No Abstracts.
    Type of Medium: Electronic Resource
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  • 46
    Electronic Resource
    Electronic Resource
    Berlin : Wiley-Blackwell
    Acta Biotechnologica 16 (1996) 
    ISSN: 0138-4988
    Keywords: Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Process Engineering, Biotechnology, Nutrition Technology
    Type of Medium: Electronic Resource
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  • 47
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    Berlin : Wiley-Blackwell
    Acta Biotechnologica 16 (1996), S. 3-17 
    ISSN: 0138-4988
    Keywords: Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Process Engineering, Biotechnology, Nutrition Technology
    Notes: Molecular evolution has recently been applied in biotechnology which consist of the development of evolutionary strategies in the design of biopolymers with predefined properties and functions. At the heart of this new technology are the in vitro replication and random synthesis of RNA or DNA molecules, producing large libraries of genotypes that are subjected to selection techniques following DARWIN's principle. By means of these evolutionary methods, RNA molecules were derived which specifically bind to predefined target molecules. Ribozymes with new catalytic functions were obtained as well as RNA molecules that are resistant to cleavage by specific RNases. In addition, the catalytic specificities of group I introns, a special class of ribozymes, were modified by variation and selection.Efficient applications of molecular evolution to problems in biotechnology require a fundamental and detailed understanding of the evolutionary process. Two basic questions are of primary importance: (i) How can evolutionary methods be successful as the numbers of possible genotypes are so large that the chance of obtaining a particular sequence by random processes is practically zero, and (ii) how can populations avoid being caught in evolutionary traps corresponding to local fitness optima? This review is therefore concerned with an abridged account of the theory of molecular evolution, as well as its application to biotechnology. We add a brief discussion of new techniques for the massively parallel handling and screening of very small probes as is required for the spatial separation and selection of genotypes. Finally, some imminent prospects concerning the evolutionary design of biopolymers are presented.
    Additional Material: 7 Ill.
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  • 48
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    Berlin : Wiley-Blackwell
    Acta Biotechnologica 16 (1996), S. 31-41 
    ISSN: 0138-4988
    Keywords: Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Process Engineering, Biotechnology, Nutrition Technology
    Notes: Toluene was anaerobically degraded by an enriched mixed culture under methanogenic conditions. The mixed culture was originally developed from cow-dung and sludge from a laboratory reactor, in which benzene was anerobically degraded by sulphate-reducing bacteria. First the mixed culture was enriched on toluene over a year with and without the use of sulphate in the medium. For the evaluation of growth-kinetic and maintenance parameters, namely μmax, Ks, kd and Yx/so, the anaerobic degradation of toluene was carried out in batch as well as in continous reactors systems. The gas volume and the methane content in the produced gas was somewhat lover than the theoretical value expected, indicating an incomplete degradation of some of the complex intermediates of the toluene degradation pathway. However, the mixed culture was able to transform 41.3% of the toluene carbon into methane.
    Additional Material: 6 Ill.
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  • 49
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    Berlin : Wiley-Blackwell
    Acta Biotechnologica 16 (1996), S. 42-42 
    ISSN: 0138-4988
    Keywords: Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Process Engineering, Biotechnology, Nutrition Technology
    Notes: No Abstracts.
    Type of Medium: Electronic Resource
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  • 50
    Electronic Resource
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    Berlin : Wiley-Blackwell
    Acta Biotechnologica 16 (1996), S. 315-319 
    ISSN: 0138-4988
    Keywords: Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Process Engineering, Biotechnology, Nutrition Technology
    Notes: Following our investigations on citrinin production by Monascus ruber in a chemically defined medium, the kinetic behaviour of this toxin in the fermenter was studied with relation to production of pigments, biomass and nutrient consumption. Showing a secondary metabolite pattern, citrinin was produced when dμ/dt and dQs/dt were ≤ 0, while a high specific production rate of pigments occurred when dμ/dt and dQs/dt were 〉 0. No trophophase-idiophase transition was detected during the cuitivation, and the behaviour of pigment production was similar to that of a primary metabolite.
    Additional Material: 2 Ill.
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  • 51
    ISSN: 0138-4988
    Keywords: Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Process Engineering, Biotechnology, Nutrition Technology
    Notes: The gene coding for the class 1 outer membrane protein from the Neisseria meningitidis strain B385 (B : 4 : P1.15) was isolated by the Polymerase Chain Reaction (PCR) and cloned into the Sma I cut M13mp 18 vector. Then, a Xba I restriction site was created by using an oligonucleotide-directed in vitro mutagenesis system at the start of the coding fragment. In order to express the protein, this fragment was fused to 180 base pairs corresponding to 60 amino acids from the N terminus of interleukin-2 under the control of the tryptophan promoter (Ptrp). The expression was confirmed by Western-blotting where the recombinant protein (PILM28) was detected by bactericidal monoclonal antibodies (MABs). The recombinant polypeptide was partially purified and used to elicit murine antibodies, able to recognize the meningococcal class 1 subtype 15.
    Additional Material: 5 Ill.
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  • 52
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    Berlin : Wiley-Blackwell
    Acta Biotechnologica 16 (1996), S. 175-183 
    ISSN: 0138-4988
    Keywords: Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Process Engineering, Biotechnology, Nutrition Technology
    Notes: Whey, hydrolyzed by using pure enzymes and enzymatic preparations, was fermented by propionic acid bacteria in batch cultures. The course of propionic acid fermentation was examined by the determination of cell dry weight, lactose utilization, production of volatile free acids and biosynthesis of vitamin B12. The production of propionic acid by Propionibacteria increased when hydrolyzed whey was used as a medium. It rose from 6.3 to 6.7 g/l in the control compared to 8.0-8.8 g/l when pepsin, papain, alcalase, or pescalase were applied as hydrolyzing agents. A significant decrease in the biosynthesis of vitamin B12 (more than 50%) was observed in all cases compared to the control (cultivation on medium without hydrolysis).
    Additional Material: 5 Ill.
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  • 53
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    Berlin : Wiley-Blackwell
    Acta Biotechnologica 16 (1996), S. 185-192 
    ISSN: 0138-4988
    Keywords: Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Process Engineering, Biotechnology, Nutrition Technology
    Notes: In this article, attention is focussed on an on-line sampling device for bioreactors and its characterization regarding sterility and response time. The integration of this device into analytical systems for biotechnology (FIA with biosensors) resulted in on-line analysis systems for bioprocess control. The new ESIP version of an in situ sampling system for bioprocess analysis produced by the EPPENDORF-NETHELER-HINZ GmbH, Germany, had a short response time of 8 min (99%), which was determined by means of conductivity measurement after an increase of the medium conductivity due to the gradual addition of KCl. Effectiveness and reliability of the module were tested by bubble point measurement resulting in a bubble point pressure of 2.1 bars. The sampling probe was tested successfully for use in a broad variety of microorganisms and cultivations.
    Additional Material: 4 Ill.
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  • 54
    ISSN: 0138-4988
    Keywords: Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Process Engineering, Biotechnology, Nutrition Technology
    Notes: No Abstracts.
    Type of Medium: Electronic Resource
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  • 55
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    Berlin : Wiley-Blackwell
    Acta Biotechnologica 16 (1996), S. 207-213 
    ISSN: 0138-4988
    Keywords: Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Process Engineering, Biotechnology, Nutrition Technology
    Notes: Optimum conditions for citric acid production by Aspergillus niger in submerged culture were established. Some correlation was found between the ability of the fungus to produce citric acid and its capability to accumulate extracellular acid phosphatase. Mutagenization and passage on sodium citrate (10-25%) medium gave rise to eleven A. niger strains able to grow when the concentration was 25%. Two mutants showed an increase in citric acid production (8.8-25.7%), accompanied by the highest activity of acid phosphatase (43.1-46.6%).
    Additional Material: 3 Tab.
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  • 56
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    Berlin : Wiley-Blackwell
    Acta Biotechnologica 16 (1996), S. 224-224 
    ISSN: 0138-4988
    Keywords: Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Process Engineering, Biotechnology, Nutrition Technology
    Notes: No Abstracts.
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  • 57
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    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 101-113 
    ISSN: 0749-503X
    Keywords: ARS ; genome analysis ; chromosome V ; physical mapping ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: We have developed a new procedure for easy and rapid identification of autonomously replicating sequences (ARSs) and have applied it to the analysis of chromosome V of Saccharomyces cerevisiae. The procedure makes use of the ordered λ phage clone bank of this chromosome that we have constructed, and includes transposition of a mini-transposon and selection of transposon-containing derivatives, isolation of their DNA and circularization at their cos-ends, transformation of yeast cells with the circularized DNA, and scoring transformation frequency. The transposon used was derived from Tn5supF, contained the yeast LEU2 gene, and was placed, together with the hyperactive transposase gene, on a mini-F plasmid for stable maintenance in Escherichia coli K-12. Sixteen regions of chromosome V showing ARS activity were identified, of which 12 were newly found in this work. Thus, the procedure will be useful for systematic genomic scale analysis of ARSs in yeast and related organisms in which ordered clone banks have been established. The average distance between adjacent ARS-containing regions was approximately 40 kb. Two-dimensional gel electrophoretic analysis of chromosome replication indicated that one of the newly identified ARSs was functional as an actual in situ replication origin, at least under the conditions employed.
    Additional Material: 5 Ill.
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  • 58
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    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 135-143 
    ISSN: 0749-503X
    Keywords: peroxisome ; Zellweger syndrome ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Peroxisome assembly in Saccharomyces cerevisiae requires the products of several genes. In this report, the PAS5 gene has been characterized. The gene is on the left arm of chromosome X and encodes a polypeptide with similarity to the mammalian peroxisome assembly factor-1 (PAF-1). Two different length transcripts are produced from the yeast PAS5 gene. The longer mRNAs encompass an open reading frame, while the shorter transcripts initiate 46-110 base pairs downstream of the first in-frame AUG. The longer transcripts are induced four-fold on medium containing fatty acids as the sole carbon source, while the shorter transcripts are induced up to ten-fold on medium containing glycerol as a carbon source. The full-length coding sequence encodes a protein with a calculated molecular weight of 30·7 kDa. A protein of 25 kDa could be translated from the shorter transcripts and would lack a very acidic domain found in the amino-terminal extension of the longer protein. The common portion of the proteins is very basic; the calculated pI of the longer polypeptide is 9·02 and that of the shorter protein is 10·06. The PAS5 GenBank accession number is M86538.
    Additional Material: 2 Ill.
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  • 59
    ISSN: 0749-503X
    Keywords: genome sequencing ; chromosome XIV ; Saccharomyces cerevisiae ; Myotonic Dystrophy Kinase ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: We have determined the complete nucleotide sequence of a 36·8 kb segment from the left arm of chromosome XIV carried by the cosmid 14-11. The sequence encodes the 5′ coding region of the PSD1 gene, the 3′ coding region of an unknown gene and 24 complete open reading frames, of which 18 correspond to new genes and six (SKO1, SCL41A, YGP1, YCK2, RPC31 and MFA2) have been sequenced previously. Of the 24 new genes, five show significant similarities to sequences present in the databanks. These include elongation factors 2 and the human myotonic dystrophy kinase. The sequence has been deposited in the EMBL databank under the Accession Number X92517.
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  • 60
    Electronic Resource
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    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996) 
    ISSN: 0749-503X
    Keywords: Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Type of Medium: Electronic Resource
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  • 61
    ISSN: 0749-503X
    Keywords: Saccharomyces cerevisiae ; ASK10 ; SKN7 ; two-component ; response regulator ; transcription factor ; U27209 ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Recent evidence has demonstrated that the yeast Skn7p appears to act as a ‘response regulator’ in a eukaryotic ‘two-component’ signal transduction pathway. A search to identify possible regulators of the SKN7 mediated ‘two- component’ regulatory system has identified Ask10p as a novel potential transcription factor. The ASK10 sequence has been deposited in GenBank with Accession Number U27209.
    Additional Material: 3 Ill.
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  • 62
    ISSN: 0749-503X
    Keywords: Saccharomyces cerevisiae ; chromosome VI ; Snf2/Rad54 helicase family ; SUP6 ; HIS2 ; CDC14 ; MET10 ; SMC2 ; QCR6 ; PHO4 ; CDC26 ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The nucleotide sequence of cosmid clone 9765, which contains 30·8 kb of the right arm of chromosome VI, was determined. Both strands were sequenced, with an average redundancy of 8·17 per base pair by both dye primer and dye terminator cycle sequencing methods. The G+C content of the sequence was found to be 40·3%. Fifteen open reading frames (ORFs) greater than 100 amino acids and one tRNA-Tyr gene (SUP6) were detected. Seven of the ORFs were found to encode previously identified genes (HIS2, CDC14, MET10, SMC2, QCR6, PH04 and CDC26). One ORF, 9765orfF010, was found to encode a new member of the Snf2/Rad54 helicase family. Three ORFs (9765orfR002, 9765orfR011 and 9765orfR013) were found to be homologous with Schizosaccharomyces pombe polyadenylate binding protein, Escherichia coli hypothetical 38·1-kDa protein in the BCR 5′ region, and transcription regulatory protein Swi3, respectively. The sequence may be found in the DDBJ, EMBL and GenBank nucleotide sequence databases under Accession Number D44602.
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  • 63
    ISSN: 0749-503X
    Keywords: S. cerevisiae ; autoselection plasmid ; flow cytometry ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Autoselection systems allow the selection of a genetically engineered population independently of the growth medium composition. The structure of a Saccharomyces cerevisiae population transformed with an autoselection plasmid, in which a carbon-source-dependent modulation of the plasmid copy number occurs, was analysed.By means of flow cytometric procedures we tested the cell viability, dynamics of growth and heterologous protein production at single cell level. Such analyses allow the identification and the tracking of a specific cellular sub-population with a higher plasmid copy number which arises after the carbon source shift. The effects of the cellular plasmid distribution on the dynamics of growth are also discussed.
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  • 64
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    Yeast 12 (1996), S. 299-306 
    ISSN: 0749-503X
    Keywords: Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
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  • 65
    ISSN: 0749-503X
    Keywords: pyruvate decarboxylase ; sugar metabolism ; Saccharomyces cerevisiae ; metabolic compartmentation ; acetyl-CoA ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: In Saccharomyces cerevisiae, the structural genes PDC1, PDC5 and PDC6 each encode an active pyruvate decarboxylase. Replacement mutations in these genes were introduced in a homothallic wild-type strain, using the dominant marker genes APT1 and Tn5ble. A pyruvate-decarboxylase-negative (Pdc-) mutant lacking all three PDC genes exhibited a three-fold lower growth rate in complex medium with glucose than the isogenic wild-type strain. Growth in batch cultures on complex and defined media with ethanol was not impaired in Pdc- strains. Furthermore, in ethanol-limited chemostat cultures, the biomass yield of Pdc- and wild-type S. cerevisiae were identical. However, Pdc- S. cerevisiae was unable to grow in batch cultures on a defined mineral medium with glucose as the sole carbon source. When aerobic, ethanol-limited chemostat cultures (D = 0·10 h-1) were switched to a feed containing glucose as the sole carbon source, growth ceased after approximately 4 h and, consequently, the cultures washed out. The mutant was, however, able to grow in chemostat cultures on mixtures of glucose and small amounts of ethanol or acetate (5% on a carbon basis). No growth was observed when such cultures were used to inoculate batch cultures on glucose. Furthermore, when the mixed-substrate cultures were switched to a feed containing glucose as the sole carbon source, wash-out occurred. It is concluded that the mitochondrial pyruvate dehydrogenase complex cannot function as the sole source of acetyl-CoA during growth of S. cerevisiae on glucose, neither in batch cultures nor in glucose-limited chemostat cultures.
    Additional Material: 6 Ill.
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  • 66
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    Yeast 12 (1996), S. 307-317 
    ISSN: 0749-503X
    Keywords: bent DNA ; Rap1p ; glycolytic enzyme gene-regulator ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: In Saccharomyces cerevisiae the GCR1 gene product is required for high-level expression of genes encoding glycolytic enzymes. In this communication, we extend our analysis of the DNA binding properties of Gcr1p. The DNA-binding domain of Gcr1p binds DNA with high affinity. The apparent dissociation constant of the Gcr1p DNA-binding domain for one of its specific binding sites (TTTCAGCTTCCTCTAT) is 2·9×10-10 M. However, competition experiments showed that Gcr1p binds this site in vitro with a low degree of specificity. We measured a 33-fold difference between the ability of specific competitor and DNA of random sequence to inhibit the formation of nucleoprotein complexes between Gcr1p and a radiolabeled DNA probe containing its binding site. DNA band-shift experiments, utilizing probes of constant length in which the positions of Gcr1p-binding sites are varied relative to the ends, indicated that Gcr1p-DNA nucleoprotein complexes contain bent DNA. The implications of these findings in terms of the combinatorial interactions that occur at the upstream activating sequence elements of genes encoding glycolytic enzymes are discussed.
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  • 67
    ISSN: 0749-503X
    Keywords: inositol ; phosphatidylinositol synthase ; Candida albicans ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Phosphatidylinositol (PI) synthase (cytidine 5′-diphospho (CDP)-1,2-diacyl-sn-glycerol:myo-inositol 3phosphatidyltransferase, EC 2.7.8.11) was isolated from the microsomal cell fraction of Candida albicans. The Triton X-100 extracted enzyme was enriched 140-fold by affinity chromatography on CDP-diacylglycerol-Sepharose. The enzyme had a pH optimum at 9·5 in glycine/NaOH buffer. It had an absolute requirement for Mg2+ or Mn2+ and was inhibited by Ca2+ and Zn2+. Maximal activity was at 0·2-0·6 mm-CDP-diacylglycerol, higher concentrations inhibited the enzyme. With 2′-deoxy-CDP-diacylglycerol as the lipid substrate, optimal activity was at 0·7 mm. The Km for myo-inositol was determined to be 0·55 mm. The optimal temperature for the PI synthase reaction was 55°C. The C. albicans PI synthase shows differences to the Saccharomyces cerevisiae enzyme, such as activation by bivalent cations, inhibition by nucleotides, temperature optimum and activation energy, but also to the human PI synthase in preference for the lipid substrates, inhibition by nucleoside monophosphates and stabilization by Mn2+ and phospholipids.
    Additional Material: 4 Ill.
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  • 68
    ISSN: 0749-503X
    Keywords: Saccharomyces cerevisiae ; genome sequencing ; chromosome XIV ; leukotriene A4 hydrolase ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: We have determined the nucleotide sequence of a 12·8 kb fragment from the left arm of chromosome XIV carried by the cosmid 14-16d. An analysis of the sequence reveals the presence of a sigma element, a pro-tRNA gene and eight open reading frames, six of which are complete. All of the eight open reading frames correspond to new genes. Of the eight new genes, two show strong similarities to a pair of new genes from chromosome IX, suggesting an ancestral duplication, and one gene encodes a protein similar to mammalian leukotriene A4 hydrolase. The sequence has been deposited in the EMBL data bank under the Accession Number X94547.
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  • 69
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    Yeast 12 (1996) 
    ISSN: 0749-503X
    Keywords: Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
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  • 70
    ISSN: 0749-503X
    Keywords: glycosylation ; dolichyl phosphoryl mannose ; dolichol ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Genomic DNAs from several fungi were screened for a homologous sequence to Saccharomyces cerevisiae DPM1, an essential gene which encodes dolichyl phosphoryl mannose synthase. The fungi examined included Aspergillus nidulans, Neurospora crassa, Schizophyllum commune and Ustilago maydis. Only U. maydis gave a significant signal after Southern hybridization using DPM1 as a probe. The Ustilago homolog was subsequently cloned and sequenced. The predicted protein of 294 amino acids has 60% identity to the S. cerevisiae protein, but lacks the putative ‘dolichol recognition sequence’. RNA of ca. 900 bp is transcribed in both yeast and filamentous cells of Ustilago. In Escherichia coli, the U. maydis sequence expressed a 35 kDa protein exhibiting dolichyl phosphoryl mannose synthase activity. The sequence was also shown to complement a haploid strain of S. cerevisiae containing a deletion of the DPM1 gene. The U. maydis sequence therefore, encodes a dolichyl phosphoryl mannose synthase that can support normal vegetative growth in S. cerevisiae. The GenBank accession number is U54797.
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  • 71
    ISSN: 0749-503X
    Keywords: Saccharomyces cerevisiae ; chromosome X ; open reading frames ; tRNA ; gene duplication ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: We have sequenced a 61,989 bp stretch located between genes RAD7 and FIP1 of Saccharomyces cerevisiae chromosome X. This stretch contains 36 open reading frames (ORFs) of at least 100 codons. Fourteen of these correspond to sequences previously published as HIT1, CDC8, YAP17, CBF1, NAT1, RPA12, CCT5, TOR1, RFC2, PEM2, CDC11, MIR1, STE18 and GRR1. The proteins deduced from four ORFs (YJR059w, YJR065c, YJR075w, YJR078w) have significant similarity to proteins of known function from yeast or other organisms, including S. cerevisiae serine/threonine-specific protein kinase, Schizosaccharomyces pombe Act2 protein, S. cerevisiae mannosyltransferase OCH1 protein and mouse indoleamine 2,3-dioxygenase, respectively. Four of the remaining 18 ORFs have similarity to proteins with unknown function, six are weakly similar to other known sequences, while another eight exhibit no similarity to any known sequence. In addition, three tRNA genes have been recognized. Three genes clustered within 22 kb (YJR059w, YJR061w and TOR1) have counterparts arranged within 15 kb on the left arm of chromosome XI. The sequence has been deposited in the Genome Sequence Data Base under Accession Number L47993.
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  • 72
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    Yeast 12 (1996), S. 877-885 
    ISSN: 0749-503X
    Keywords: Saccharomyces cerevisiae ; chromosome sequencing ; chromosome XV ; MGM1 ; STE4 ; CDC44 ; STE13 ; RPB8 ; MIP1 ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: We have determined the nucleotide sequence of a cosmid (pEOA423) from chromosome XV of Saccharomyces cerevisiae. Analysis of the 33,173 bp sequence reveals the presence of 20 putative open reading frames (ORFs). Five of them correspond to previously known genes (MGM1, STE4, CDC44, STE13, RPB8). The previously published nucleotide sequences are in perfect agreement with our sequence except for STE4 and MGM1. In the latter case, 59 amino acids were truncated from the published protein at its N-terminal end due to a frameshift. The putative translation products of six other ORFs exhibit significant homology with protein sequences in public databases: O50 03 and O50 17 products are homologs of the ANC1 and MIP1 proteins of S. cerevisiae, respectively; O50 05 product is similar to that of a protein of unknown function from Myxococcus xanthus; O50 12 product is probably a new ATP/ADP carrier; O50 13 product shows homology with group II tRNA synthetases; and the O50 16 product exhibits strong similarity with the N-terminal domain of the NifU proteins from several prokaryotes. The remaining nine ORFs show no significant similarity. Among these, two contiguous ORFs (O50 19 and O50 20) are very similar to each other, suggesting an ancient tandem duplication. The 33,173 bp sequence has been submitted to EMBL (Accession Number: X92441).
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  • 73
    ISSN: 0749-503X
    Keywords: Saccharomyces cerevisiae ; chromosome VII ; SEC27 ; SSM1b ; S-adenosylmethionine-dependent enzyme ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The nucleotide sequence of a fragment from the left arm of Saccharomyces cerevisiae chromosome VII has been determined. Analysis of the 14,607 bp DNA segment reveals nine open reading frames (ORFs) longer than 300 bp. G2827 is the SEC 7 gene, an essential coatomer complex subunit. G2834 encodes SSM1b, a ribosomal protein. The G2838 product shows homology to hypothetical yeast proteins, YIF0 and YE09, of unknown function. The G2830 product shows homology with the cell division protein FtsJ from Escherichia coli, with two hypothetical proteins from yeast, YCF4 and YBR1, and with R74.7, a hypothetical protein from Caenorhabditis elegans. Two of the ORFs are completely internal to longer ones and a third is partially embedded in G2850. The remaining ORFs give no significant homology with proteins in the databases. The sequence has been deposited at the EMBL database under Accession Number X92670.
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  • 74
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    Yeast 12 (1996), S. 859-868 
    ISSN: 0749-503X
    Keywords: gene disruption ; Saccharomyces cerevisiae ; yeast ; function analysis ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: We describe a new procedure for the generation of plasmids containing a large promoter and terminator region of a gene of interest, useful for gene disruption. In a two-step polymerase chain reaction (PCR), a fragment, corresponding to the terminator and promoter regions separated by a 16 bp sequence containing a rare restriction site (e.g. AscI), is synthesized (T-P fragment). This PCR fragment is cloned in vectors presenting a rare blunt-end cloning site and a yeast marker for selection in Saccharomyces cerevisiae (TRP1, HIS3 and KanMX). The final plasmids are used directly for gene disruption after linearization by the enzyme (e.g. AscI) specific for the rare restriction site. This approach was used to disrupt three open reading frames identified during the sequencing of COS14-1 from chromosome XIV of S. cerevisiae.
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  • 75
    ISSN: 0749-503X
    Keywords: Saccharomyces cerevisiae ; genome sequencing ; chromosome IV ; DOA4 ; UBC5 ; UBC3 ; DNA52 ; ribosomal protein gene ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: We have sequenced a region containing 32.5 kb of the right arm of chromosome IV of Saccharomyces cerevisiae. Twenty open reading frames (ORFs) greater than 100 amino acids could be identified in this region. Six ORFs correspond to known yeast genes, including DOA4, UBC5 and UBC3, the gene products of which are involved in ubiquitin metabolism. UBC5 is preceded by the two tRNA genes tRNA-Arg2 and tRNA-Asp. Six genes were discovered with homologies to non-yeast genes or with homologies to other yeast ORFs. One of these could be identified as ribosomal protein gene RPS13. The putative function of eight ORFs remains unclear because comparison to different DNA or protein databases revealed no significant patterns. The sequence from cosmid 2F21 was obtained entirely by a combined subcloning and walking primer strategy, and has been deposited in the EMBL data library under Accession Number X84162.
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  • 76
    ISSN: 0749-503X
    Keywords: Saccharomyces cerevisiae ; chromosome VI ; SEC4 ; MSH4 ; TYA ; TYB ; SPB4 ; DEG1 ; NIC96 ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Plasmid clone gapB and lambda phage clone 4682, which contain fragments of Saccharomyces cerevisiae chromosome VI, were analysed. A 23 kb sequence was determined and ten open reading frames (ORFs) were revealed. Among them, five ORFs were identical to five yeast genes (SEC4, MSH4, SPB4, DEG1 and NIC96), two were identical to transposable elements (TYA and TYB), one (gapBorfF003) was highly homologous to a yeast expressed sequence tag, and another (4682orfF002) was predicted to be a nuclear protein. Sequence data have been submitted to DDBJ/EMBL/GenBank data library under Accession Number D44604 (clone gapB) and D44600 (clone 4682), respectively.
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  • 77
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    Yeast 12 (1996) 
    ISSN: 0749-503X
    Keywords: Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
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  • 78
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    Yeast 12 (1996), S. 93-100 
    ISSN: 0749-503X
    Keywords: Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
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  • 79
    ISSN: 0749-503X
    Keywords: Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
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  • 80
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    Yeast 12 (1996), S. 1-16 
    ISSN: 0749-503X
    Keywords: Saccharomyces cerevisiae ; lysosome ; sorting ; proteolytic activation ; endocytosis autophagocytosis ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The yeast vacuole, which is equivalent to the lysosome of higher eukaryotes, is one of the best characterized degradative organelles. This review describes the biosynthesis and function of yeast vacuolar proteases. Most of these enzymes are delivered to the vacuole via the early compartments of the secretory pathway and the endosome, while one of them is directly imported from the cytoplasm. The proteases are synthesized as precursors which undergo many post-translational modifications before the final active form is generated. Proteolytic activation by removal of propeptides is performed by proteinase A and/or proteinase B in an intricate cascade-like fashion. Recent developments in the analysis of the functions of vacuolar proteolysis are described. Substrates of the vacuolar proteases are mostly imported via endocytosis or autophagocytosis, and vacuolar proteolysis appears to be mainly important under nutritional stress conditions and sporulation.
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  • 81
    ISSN: 0749-503X
    Keywords: genome sequencing ; Saccharomyces cerevisiae ; chromosome VII ; CEG1 ; SOH1 ; DnaJ ; SCS3 ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: We report the sequence analysis of a 10 kb DNA fragment of Saccharomyces cerevisiae chromosome VII. This sequence contains four complete open reading frames (ORFs) of greater than 100 amino acids. There are also two incomplete ORFs flanking the extremes: one of these, G2868, is the 5′ part of the SCS3 gene (Hosaka et al., 1994). ORFs G2853 and G2856 correspond to the genes CEG1, coding for the alfa subunit of the mRNA guanylyl transferase and a 3′ gene of unknown function previously sequenced (Shibagaki et al., 1992). G2864 is identical to SOH1 also reported (Fan and Klein, 1994). The translated sequence of G2861 is similar to the human dnaJ homolog. The nucleotide sequence reported here has been entered in the EMBL Data Library under the Accession Number X87252.
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  • 82
    ISSN: 0749-503X
    Keywords: Saccharomyces cerevisiae ; gene disruption ; PCR-mediated disruption ; uraduction ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Gene disruption is an important method for genetic analysis in Saccharomyces cerevisiae. We have designed a polymerase chain reaction-directed gene disruption cassette that allows rapid disruption of genes in S. cerevisiae without previously cloning them. In addition, this cassette allows recycling of URA3, generating gene disruptions without the permanent loss of the ura3 marker. An indefinite number of disruptions can therefore be made in the same strain.
    Additional Material: 3 Ill.
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  • 83
    ISSN: 0749-503X
    Keywords: Candida albicans ; alcohol dehydrogenase ; gene regulation ; messenger RNA ; carbon utilization ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The Candida albicans ADH1 gene encodes an alcohol dehydrogenase which is immunogenic during infections in humans. The ADH1 gene was isolated and sequenced, and the 5′- and 3′-ends of its mRNA were mapped. The gene encodes a 350 amino acid polypeptide with strong homology (70·5-85·2% identity) to alcohol dehydrogenases from Saccharomyces cerevisiae, Kluyveromyces lactis and Schizosaccharomyces pombe. The cloned C. albicans ADH1 gene was shown to be functional through complementation of adh mutations and efficient production of active alcohol dehydrogenase in S. cerevisiae. Northern analysis of C. albicans RNA revealed that ADH1 mRNA levels were regulated in response to carbon source and during batch growth. During growth on glucose, ADH1 mRNA levels rose to maximum levels during late exponential growth phase and declined to low levels in stationary phase. The ADH1 mRNA was relatively abundant during growth on galactose, glycerol, pyruvate, lactate or succinate, and less abundant during growth on glucose or ethanol. Alcohol dehydrogenase levels did not correlate closely with ADH1 mRNA levels under the growth conditions studied, suggesting either that this locus is controlled at both transcriptional and post-transcriptional levels, or that other differentially regulated ADH loci exist in C. albicans.
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  • 84
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    Yeast 12 (1996), S. 237-240 
    ISSN: 0749-503X
    Keywords: electrophoretic karyotype ; linkage group ; Saccharomycodes ludwigii ; tetrad analysis ; yeast ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Tetrad distributions for 108 different gene pairs in 1346 asci of 113 diploids heterozygous for various combinations of 24 genes in Saccharomycodes ludwigii were investigated. Tetratype tetrads occurred only rarely and the 24 genes tested were classified into seven linkage groups. Electrophoretic karyotypes of three independent strains of S'codes ludwigii showed seven bands of chromosome-sized DNA having molecular sizes of 0·8 to 2·3 Mb with strain-specific polymorphic chromosomal DNAs as determined based on their migration distances.
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  • 85
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    Yeast 12 (1996), S. 289-295 
    ISSN: 0749-503X
    Keywords: genome sequencing ; Saccharomyces cerevisiae ; chromosome XIV ; right telomere ; sub-telomeric repeats ; mannitol dehydrogenase homolog ; YCR007/YKL219-like ; PAU1- like ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: We report the complete sequence of a 9·2 kb fragment next to and including the right telomere of Saccharomyces cerevisiae chromosome XIV. Four open reading frames (ORFs) longer than 100 amino acids were observed in the sequenced segment. One ORF (378 codons) does not show any significant homology with proteins in the databases and corresponds to a putative new gene. Two ORFs are almost identical to the known YCR007/YKL219 and PAU1-like hypothetical protein families already identified on several S. cerevisiae chromosomes. These ORFs, whose function is unknown, are generally associated with sub-telomeric regions of chromosomes. The fourth one shows significant identities with bacterial mannitol dehydrogenases. It could be a yeast gene implicated in the metabolism of mannitol (or a related substrate). The sequence has been deposited in the EMBL data library under Accession Number X86790.
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  • 86
    ISSN: 0749-503X
    Keywords: Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Additional Material: 1 Tab.
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  • 87
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    Yeast 12 (1996) 
    ISSN: 0749-503X
    Keywords: Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
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  • 88
    ISSN: 0749-503X
    Keywords: transport ; glucose uptake ; Saccharomyces cerevisiae ; yeast ; rapid kinetics ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Glucose uptake in Saccharomyces cerevisiae is believed to consist of two kinetically distinguishable components, the affinity of which is modulated during growth on glucose. It has been reported that triple hexose-kinase deletion mutants do not exhibit high-affinity glucose uptake. This raises the question of whether and how high-affinity glucose uptake is related to the presence of glucose-phosphorylating enzymes. In this study the kinetics of glucose uptake in both wild-type cells and cells of hexose-kinase deletion mutants, grown on either glycerol or galactose, were determined using a rapid-uptake method. In wild-type cells glucose uptake measured over either 5 s or 200 ms exhibited high affinity. In contrast, in cells of hexose-kinase deletion mutants the apparent affinity of glucose uptake was dependent on the time scale during which uptake was measured. Measurements on the 5-s scale showed apparent low-affinity uptake whereas measurements on the 200-ms scale showed high-affinity uptake. The affinity and maximal rate of the latter were comparable to those in wild-type cells.Using a simple model for a symmetrical facilitator, it was possible to simulate the experimentally determined relation between apparent affinity and the time scale used.The results suggest that high-affinity glucose transport is not necessarily dependent on the presence of glucose-phosphorylating enzymes. Apparent low-affinity uptake kinetics can arise as a consequence of an insufficient rate of removal of intracellular free glucose by phosphorylation.This study underlines the need to differentiate between influences of the translocator and of metabolism on the apparent kinetics of sugar uptake in yeast.
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  • 89
    ISSN: 0749-503X
    Keywords: Saccharomyces cerevisiae ; chromosome XIV ; RHO2 ; TOP2 ; MKT1 ; END3 ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: We report the DNA sequence of a 17 933 bp fragment from the left arm of chromosome XIV of Saccharomyces cerevisiae. Analysis of the sequence reveals the presence of ten open reading frames (ORFs) larger than 100 codons. Four of these were previously identified as genes RHO2, TOP2, MKT1 and END3. Additionally, the NH2 end coding region of PMS1 is found in the 3′ end of the sequence. No significant homology to any known protein has been found for the other five ORFs. The nucleotide sequence has been deposited at EMBL, with Accession Number X89016.
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  • 90
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    Yeast 12 (1996), S. 501-504 
    ISSN: 0749-503X
    Keywords: Candida albicans ; chitinase ; yeast ; nucleotide sequencing ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Here we report the complete nucleotide sequence of a third chitinase gene (CHT1) from the dimorphic human pathogen Candida albicans. The deduced amino acid (aa) sequence of Cht1 consists of 416 aa and displays 36% protein sequence similarity to chitinases Cht2 and Cht3, from C. albicans. Interestingly the domain structure of Cht1 is truncated when compared to the other chitinases of C. albicans and lacks a Ser/Thr-rich region. The sequence data will appear in the GenBank Nucleotide Sequence Data Library under the accession number U36490.
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  • 91
    ISSN: 0749-503X
    Keywords: Biosurfactant ; glycolipid ; cytochrome P450 ; Candida apicola ; alkane assimilation ; fatty acid hydroxylation ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Candida apicola belongs to a group of yeasts producing high amounts of surface-active extracellular glycolipids consisting of sophorose and long-chain-ω- and (ω-1)-hydroxy fatty acids. The involvement of cytochrome P450 in the synthesis of sophorose lipid by the hydroxylation of long-chain fatty acids was suggested from a simultaneous increase of cellular P450 content. Hydroxylation studies indicated the existence of multiple P450 forms capable of hydroxylating not only long-chain fatty acids, but also n-alkanes.In this report, two different P450 DNA fragments amplified in a polymerase chain reaction with heterologous primers and chromosomal DNA of Candida apicola were used as homologous probes for the isolation of full-length clones from a genomic library. The open reading frames of both genes encode proteins of 519 amino acids with calculated molecular weights of 58,656 and 58,631, respectively, that contain N-terminal membrane anchor sequences and hallmark residues, in common with other eukaryotic P450s. The deduced amino acid sequences of the C. apicola P450 genes are 84·4% identical. They share 34·5 to 44·1% identity with the proteins of the yeast family CYP52 and about 25% identity with fatty acid hydroxylases of higher eukaryotes (family CYP4A) and of Bacillus megaterium (CYP102). Southern hybridization experiments revealed the existence of further P450-related genes in C. apicola. According to the P450 nomenclature system, the cloned genes were named CYP52E1 and CYP52E2, establishing a new subfamily in yeast family CYP52. The sequences were deposited in the EMBL/GenBank Library under the Accession Numbers X76225 and X87640.
    Additional Material: 6 Ill.
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  • 92
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996) 
    ISSN: 0749-503X
    Keywords: Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Type of Medium: Electronic Resource
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  • 93
    ISSN: 0749-503X
    Keywords: CEN12 ; DNM1 ; MMM1 ; DRS1 ; SOF1 ; SCD25 ; DPS1/ATS/APSG ; TGL1 ; hMRP1 ; hCFTR ; yeast ; cystic fibrosis ; multidrug resistance ; Pumilio ; MPT5 ; HTR1 ; YGL3 ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: In the framework of the European Union BIOTECH project for systematically sequencing the Saccharomyces cerevisiae genome, we determined the nucleotide sequence of a 43·7 kb DNA fragment spanning the centromeric region of chromosome XII. A novel approach was the distribution of sublibraries prepared by the DNA coordinator (J. Hoheisel, Heidelberg, FRG), using a new hybridization-based DNA mapping method, in order to facilitate ordered sequencing. The sequence contains 22 open reading frames (ORFs) longer than 299 bp, including the published sequences for ATS/DPS1, SCD25, SOF1, DRS1, MMM1, DNM1 and the centromeric region CEN12. Five putative ORF products show similarity to known proteins: the leucine zipper-containing ABC transporter L1313p to the yeast Ycf1p metal resistance protein, to the yeast putative ATP-dependent permease Yhd5p, to the yeast putative proteins Yk83p and Yk84p, to the human cystic fibrosis transmembrane conductance regulator protein (hCFTR) and to the human multidrug resistance-associated protein hMRP1; L1325p to the Drosophila melanogaster Pumilio protein, to the putative yeast regulatory protein Ygl3p and to the yeast protein Mpt5p/Htr1p; L1329p to human lipase A and gastric lipase, to rat lingual lipase and to the putative yeast triglyceride lipase Tgl1p; L1341p to the putative yeast protein Yhg4p; and the leucine zipper-containing L1361p to the two yeast proteins 00953p and Ym8156.08p and to the Arabidopsis thaliana protein HYP1. Eight ORFs show no homology to known sequences in the database, three small ORFs are internal and complementary to larger ones and L1301 is complementary overlapping the ATS/DPS1 gene. Additionally three equally spaced ARS consensus sequences were found. The nucleotide sequence reported here has been submitted to the EMBL data library under the accession number X91488.
    Additional Material: 2 Ill.
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  • 94
    ISSN: 0749-503X
    Keywords: Saccharomyces cerevisiae ; chromosome XV ; sugar transporter ; carboxypeptidase ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: We report the sequence of a 15·5 kb DNA segment located near the left telomere of chromosome XV of Saccharomyces cerevisiae. The sequence contains nine open reading frames (ORFs) longer than 300 bp. Three of them are internal to other ones. One corresponds to the gene LGT3 that encodes a putative sugar transporter. Three adjacent ORFs were separated by two stop codons in frame. These ORFs presented homology with the gene CPS1 that encodes carboxypeptidase S. The stop codons were not found in the same sequence derived from another yeast strain. Two other ORFs without significant homology in databases were also found. One of them, O0420, is very rich in serine and threonine and presents a series of repeated or similar amino acid stretches along the sequence. The nucleotide sequence has been deposited in the EMBL data library under Accession Number X89715.
    Additional Material: 4 Ill.
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  • 95
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 631-640 
    ISSN: 0749-503X
    Keywords: silencing ; mating type ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: A novel Saccharomyces cerevisiae gene, HST1, was identified from among anonymous cDNAs and the complete corresponding genomic clone was isolated and sequenced. HST1 is very closely related to SIR2, showing 71% sequence identity over 84% of its length. Polymerase chain reaction with degenerate primers on S. cerevisiae DNA identified three additional SIR2-related genes designated HST2, HST3 and HST4. The sequences of HST2, HST3 and HST4 correspond to sequences previously released by the S. cerevisiae genome sequencing project as U33335, NCBI gi:965078; X87331, NCBI gi:829135; and Z48784, YD9346.03, respectively. Disruption of HST1 has shown no phenotype with respect to mechanisms in which SIR2 has a role, namely, regional silencing of HMLα, or in rDNA recombination. The sequence of HST1 has been deposited in the DDBJ, EMBL and GenBank at NCBI database under Accession Number L47120.
    Additional Material: 4 Ill.
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  • 96
    ISSN: 0749-503X
    Keywords: genome sequencing ; Saccharomyces cerevisiae ; chromosome X ; VPS35 ; INO1 ; SnR128 ; SnR190 ; MP12 ; YAK1 ; RPB4 ; YUR1 ; TIF2 ; MRS3 ; URA2 ; RSP25 homologue ; leucine zippers ; membrane proteins ; glycogenin glycosyltransferase ; human phospholipase D ; chromosome XI ; gene cluster translocation/recombination ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The complete nucleotide sequence of a 40·7 kb segment about 130 kb from the left end of chromosome X of Saccharomyces cerevisiae was determined from two overlapping cosmids. Computer analysis of that sequence revealed the presence of the previously known genes VPS35, INO1, SnR128, SnR190, MP12, YAK1, RPB4, YUR1, TIF2, MRS3 and URA2, three previously sequenced open reading frames (ORFs) of unknown function 5′ of the INO1, 5′ of the MP12 and 3′ of the URA2 genes and 13 newly identified ORFs. One of the new ORFs is homologous to mammalian glycogenin glycosyltransferases and another has similarities to the human phospholipase D. Some others contain potential transmembrane regions or leucine zipper motifs. The existence of yeast expressed sequence tags for some of the newly identified ORFs indicates that they are transcribed. A cluster of six genes within 10 kb (YUR1, TIF2, two new ORFs, an RSP25 homologue and MRS3) have homologues arranged similarly within 28·5 kb on the right arm of chromosome XI. The sequence has been deposited in the EMBL data library under Accession Number X87371.
    Additional Material: 6 Ill.
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  • 97
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 731-740 
    ISSN: 0749-503X
    Keywords: oscillation ; glycolysis ; yeast ; Saccharomyces cerevisiae ; intracellular metabolites ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: In a population of intact cells of the yeast Saccharomyces cerevisiae the dynamics of glycolytic metabolism were investigated under the condition of sustained oscillations. At 5-s intervals cells were quenched in -40°C methanol, extracted and the intracellular concentrations of glycolytic metabolites, adenine nucleotides and phosphate were analysed. Oscillations were found for the glycolytic intermediates glucose 6-phosphate, fructose 6-phosphate and fructose 1,6-bisphosphate. At variance with earlier reports on transient glycolytic oscillations, some intermediates further down the glycolytic pathway did not oscillate significantly, even though NADH did. In addition, the adenylate energy charge and the free energy of ATP hydrolysis oscillated significantly. Dynamic coupling through the latter may be responsible for this effective compartmentation of glycolytic dynamics.
    Additional Material: 6 Ill.
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  • 98
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 723-730 
    ISSN: 0749-503X
    Keywords: Δ-9 fatty acid desaturase ; cryptococcus curvatus ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The oleaginous yeast Cryptococcus curvatus is of industrial interest because it can accumulate triacylglycerols up to 60% of the cell dry weight. We are aiming at genetic modification of fatty acid biosynthesis for the production of tailor-made triacylglycerols in C. curvatus. As a first step in the development of a transformation and expression system a gene encoding the Δ-9 fatty acid desaturase of C. curvatus (CBS 570) was cloned. The 1470 bp gene encodes a protein of 493 amino acids with a calculated molecular mass of 55 kDa. The gene shows strong similarity to previous cloned Δ-9 desaturase genes from rat and Saccharomyces cerevisiae, 62 and 72%, respectively. Expression of the Δ-9 desaturase gene was studied. Supplementation of the growth medium with oleic acid (C18:1(c9)) showed a strong repression (90%) on the mRNA level, while supplementation with petroselinic acid (C18:1(c6)) had no effect on the amount of mRNA.
    Additional Material: 6 Ill.
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  • 99
    ISSN: 0749-503X
    Keywords: ypt/rab-like proteins ; GTPases ; zinc-finger proteins ; duplications ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: A genomic clone of 7676 bp designated B22 from Saccharomyces cerevisiae has been sequenced. The 5′ end matches the previously described gene, VAM7, and the 3′ end matches the previously described gene, SPM2, both of which have been assigned to the left arm of chromosome VII. The intergenic region contains three transcribed open reading frames (ORFs). The first is related to an uncharacterized ORF of Bacillus subtilis and more weakly to MesJ in Escherichia coli; this is found as a single transcript of 1·1 kb by Northern blotting. The second ORF encodes a small ras-like GTPase of 222 residues with strong homology to yeast Ypt8p and to mammalian Rab11; this is found as a single transcript of 1·1 kb by Northern blotting. The third ORF generates a transcript of 1·6 kb and encodes a protein of 382 residues including a perfect match to the consensus sequence of a C2H2 zinc finger domain; it shares a strong homology with yeast Mig1p and Cre-A from Aspergillus, Emericella and E. coli. This ORF also has a striking similarity to a putative 43 kDa zinc finger protein encoded by an ORF (YEL8) immediately downstream of YPT8, raising the possibility that a region between VAM7 and SPM2 on chromosome VII arose as a duplication of the YPT8-YEL8 region of chromosome V, followed by a translocation. The sequence of the clone described has been deposited with the GenBank database under Accession Number U33754.
    Additional Material: 8 Ill.
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  • 100
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 12 (1996), S. 809-813 
    ISSN: 0749-503X
    Keywords: Saccharomyces cerevisiae ; 2μm plasmid ; molecular evolution ; homeologous recombination ; Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Industrial yeast strains carry one of two homeologous 2μm plasmids designated as type-1 or type-2. The 2μm plasmid, Scp1, found in common laboratory strains of Saccharomyces cerevisiae is considered a type-2 plasmid, since the ori, STB, RAF and REP1 loci and intergenic sequences of the right-unique region of Scp1 are homologous to the corresponding loci in industrial strain type-2 plasmids. However, within both its 599 bp inverted repeats Scp1 has 142-bp sequences homologous to the bakers' yeast type-1 plasmid. DNA sequence analyses and oligonucleotide hybridizations indicate that the 142-bp insertion in Scp1 was probably due to homeologous recombination between type-1 and type-2 plasmids. These results suggest that some of the plasmid and chromosomal sequence polymorphisms seen in laboratory yeast strains result from homeologous recombination in their ancestral breeding stock.
    Additional Material: 3 Ill.
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