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  • 1995-1999  (60)
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  • 1998  (60)
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  • Life Sciences
  • 1
    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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  • 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
    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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  • 4
    ISSN: 0138-4988
    Keywords: Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Process Engineering, Biotechnology, Nutrition Technology
    Notes: The attachment, growth and product synthesis of non-flocculating Zymomonas mobilis cell, fixed in stainless steel wire spheres (WS), were investigated. The carrier surface was activated by treatment with titanium (IV) chloride (TiCl4) and γ-aminopropyltriethoxysilane (AS) in an attempt to raise the efficiency in the immobilization of the cells. System productivity for ethanol and levan production, using cells immobilized on a modified stainless steel in the batch fermentation of a sucrose medium, rose as a result of increased biomass compared to the productivity of cells fixed on untreated (control) metal surfaces. Stabilized ethanol synthesis was demonstrated in the course of four cycles (each cycle 48 h) of repeated fermentations with a stainless steel carrier treated with AS, and three cycles when TiCl4 was used. Levan synthesis decreased after three cycles with cells immobilized on a silanized surface. System productivity for ethanol and levan production after the fourth cycle in experiments with TiCl4-activated, silanized and unmodified carriers were Qeth = 1.01, 1.06 and 0.27 g/l × h; Qlev = 0.32, 0.29 and 0.12 g/l × h, respectively. However, the specific productivity of biomass for product synthesis was higher in fermentation systems with untreated stainless steel surfaces, probably due to some loss of physiological activity of cells attached to a modified carrier. Investigations of throughly washed activated stainless steel wire surfaces, by scanning electron microscopy after immobilization, showed significant attachment of cells to the carriers. A polymer layer covered the wire surface treated with TiCl4 after fermentations. This may be explained as the binding of extracellular polysaccharide, such as the fructose-polymer levan and yeast extract components, to the modified support via chelation. After four fermentations, craters and holes in the polymer layer were evident, probably as a result of CO2 formation. A small number of cells appeared on this layer. In view of the good ethanol formation during all fermentation cycles, it is probably that active Z. mobilis cells remained under the polymer layer. Wire treatment with AS resulted in the formation of long filamentous cells during fermentation and some disturbance of cellular fission. This may be partly explained by strong electrostatic interactions between the positively charged carrier surface and the predominately negatively charged surface of Z mobilis cells. However, this did not significantly affect other cellular functions. The surface of the wire treated with AG was practically without a polymer layer.
    Additional Material: 6 Ill.
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  • 5
    Electronic Resource
    Electronic Resource
    Berlin : Wiley-Blackwell
    Acta Biotechnologica 18 (1998), S. 109-122 
    ISSN: 0138-4988
    Keywords: Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Process Engineering, Biotechnology, Nutrition Technology
    Notes: In fixed-bed bioreactors, the influence of the oxygen content in the inlet air on the biodegradation of diesel fuel in unsaturated soil/compost mixtures was analyzed at 30°C over a period of 7 weeks. Firstly, a wide range from 0 to 80 vol.% O2 was investigated. Subsequently, the range below 5 vol.% O2 was examined more closely. Over the whole test period of seven weeks, no significant influence of oxygen could be observed above 1 vol. % O2 in the inlet air - either on the decrease of the total contaminants or on the total mineralization. Anaerobic conditions should be avoided for the degradation of diesel fuel. During the test period, the courses of CO2 production varied significantly depending on oxygen supply. Furthermore, a model was developed to estimate the total mineralization as a function of oxygen supply. More investigations are recommended in order to test this model for practical application.
    Additional Material: 5 Ill.
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  • 6
    Electronic Resource
    Electronic Resource
    Berlin : Wiley-Blackwell
    Acta Biotechnologica 18 (1998), S. 167-174 
    ISSN: 0138-4988
    Keywords: Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Process Engineering, Biotechnology, Nutrition Technology
    Notes: Zymomonas mobilis strain 113 “S” produces levan - an extracellular, viscous, biologically active, non-toxic fructose polymer with a unique structure and extraordinary properties. This polysaccharide was isolated at two different degrees of purity by alcohol precipitation from aqueous solutions and was characterized with respect to some rheological properties and stability of viscous solutions.The effects of temperature, pH and salt concentration on the viscosity of 1-3% levan solutions were examined. The viscosity of levan solutions was found to be quite stable and reversible at room temperature over a wide range of pH from 4 to 11. The viscosity was slightly affected by increased salt concentration. Levan solutions were rather stable at high temperatures (up to 70°C, 1 h, pH 6), where the viscosity could be almost completerly restored (up to 80-100%). Therefore, the degradation of the polymer structure under these conditions is probably insignificant. Temperatures of 70-100°C with a pH of less than 3.5 caused irreversible degradation of the levan structure.The above-mentioned properties of levan, obtained from Zymomonas mobilis 113 “S”, demonstrated the potential for the development of various therapeutic forms of pharmacologically-active levan and their application in medicine as well as in the food and other industries.
    Additional Material: 3 Ill.
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  • 7
    Electronic Resource
    Electronic Resource
    Berlin : Wiley-Blackwell
    Acta Biotechnologica 18 (1998), S. 176-176 
    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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  • 8
    Electronic Resource
    Electronic Resource
    Berlin : Wiley-Blackwell
    Acta Biotechnologica 18 (1998), S. 179-188 
    ISSN: 0138-4988
    Keywords: Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Process Engineering, Biotechnology, Nutrition Technology
    Notes: The extractive purification of peroxidase from Armoracia rusticana roots and Glycine max seed coats in temperature-induced and affinity microsphere-containing aqueous two-phase systems was stuied. The extractive purification of peroxidase from Glycine max seed coats was carried out in a temperature-induced aqueous two-phase system formed by Triton X-45, Triton X-100 and sodium acetate at pH 5.5 A 99% yield with a 6-fold purification factor was obtained. When the clear top phase was subjected to concanavalin-A affinity chromatography, the purification factor rose to 41 and the yield dropped to 28%.A two-step purification process for peroxidase from Armoracia rusticana roots was developed by adding concanavalin-A affinity microspheres to a PEG/phosphate aqueous two-phase system. The method allows a 60% recovery of high purity peroxidase (1,860 guaiacol units per mg). A lower recovery rate and degree of purification of this enzyme was achieved after temperature-induced aqueous two-phase partition or acetone precipitation and concanavalin-A affinity column chromatography.
    Additional Material: 1 Ill.
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  • 9
    Electronic Resource
    Electronic Resource
    Berlin : Wiley-Blackwell
    Acta Biotechnologica 18 (1998), S. 189-200 
    ISSN: 0138-4988
    Keywords: Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Process Engineering, Biotechnology, Nutrition Technology
    Notes: Endo-β-glucanase (endo-β-1,4-glucano-glucanase EC 3.2.1.4), isolated from Trichoderma reesei, was immobilized in calcium alginate beads, retaining 75% of its original activity. The polyanionic moiety surrounding the immobilized enzyme displaced the pH-activity profile to alkaline regions with respect to that of the free enzyme. The enzyme was inhibited by carboxymethylcellulose, but this inhibition appeared to be decreased by immobilizatíon. The enzyme immobilized in alginate beads showed a Km value (1.02% w/v) lower than that of the enzyme (1.31%). The apparent Vmax of immobilized cellulase preparations (238.3 μmol glucose/ml × h) decreased by a factor of 0.59 with respect to that of the soluble enzyme. The optimum temperature (60°C) of the free and entrapped enzymes remained unaltered. In contrast, the half-life of the endoglucanase immobilized in calciumalginate beads was 4.6 h at 55°C and 5.4 h at 60°C, while that of the free enzyme was 3.0 h at 55°C and 1.2 h at 60°C. A technological application of the immobilized enzymes was tested using wheat straw as a source of fermentable sugars. The hydrolytic degradation of straw, by means of a crude extract of free and immobilized cellulases and β-glucosidase, released a large amount of reducing sugars from wheat straw after 48 h (between 250-720 mg glucose/g straw), carrying out more than a 90% saccharification. A mixture of immobilized β-glucosidase and free cellulases maintained 80% of the activity of the soluble counterparts, and the co-immobilization of both types of enzymes reduced by hydrolytic efficiency to half.
    Additional Material: 5 Ill.
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  • 10
    Electronic Resource
    Electronic Resource
    Berlin : Wiley-Blackwell
    Acta Biotechnologica 18 (1998), S. 201-229 
    ISSN: 0138-4988
    Keywords: Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Process Engineering, Biotechnology, Nutrition Technology
    Notes: The specific aspects of airlift reactors emphasizing their function relevance to particular application as bioreactors are presented.The two main groups of airlift reactors - external-loop and concentric-tube reactors - were investigated on a pilot-plant scale with regard to their performance during the cultivation of unicellular and filamentous microorganisms which produce Bacitracin, Cephalosporin C and Nystatin. Some results were compared to those obtained in conventional stirred tank bioreactors. The comparison was carried out based on physical properties (oxygen transfer rate (OTR), volumetric mass transfer coefficient (kLa) and efficiency of oxygen transfer (EO2)), cell mass, productivity and substrate consumption, secondary metabolite production, and efficiency of the product formation with regard to the specific power input.It was shown that B. licheniformis, C. acremonium and S. noursei fermentations occurred similarly to those performed in stirred vessels, proving that the capacity of the airlift bioreactors surpassed the problems which arise from the morphology and rheology of the broths. From the chemical engineering point of view, it was obvious that the primary tasks of a bioreactor (uniform distribution of microorganisms and nutrients over the entire fermenter volume, appropriate supply of biomass with nutrients and oxygen) were fulfilled by the airlift bioreactors tested. In addition, the efficiency of oxygen transfer (OTR referred to power input) in the airlift fermenters proved to be about 38% higher than in the stirred tank bioreactors (expressed as average values), while the sorption efficiency (OTR referred to antibiotic production) was found to be 22% greater in the airlift system than in an STR.Therefore, the biosyntheses were performed with about a 30-40% increase in energy efficiency and energy savings compared to the conventional system.Moreover, the lack of mechanical devices in the airlift system provides greater safety and a gentler environment for the cultivation of microorganisms.
    Additional Material: 18 Ill.
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  • 11
    Electronic Resource
    Electronic Resource
    Berlin : Wiley-Blackwell
    Acta Biotechnologica 18 (1998), S. 231-240 
    ISSN: 0138-4988
    Keywords: Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Process Engineering, Biotechnology, Nutrition Technology
    Notes: In various bacterial strains belonging to the β-subdivision of proteobacteria which are capable of degrading chlorinated monoaromatic compounds, chlorocatechol 1,2-dioxygenase genes were detected by PCR and Southern hybridization. Using PCR primers derived from the conserved sequence motifs of chlorocatechol 1,2-dioxygenase genes tfdC, clcA and tcbC, PCR products of the expected size were obtained with the test strains, but not with negative control strains. The specificity of the PCR products was verified by hybridization using an oligonucleotide probe for an internal sequence motif which is evolutionarily conserved among chlorocatechol 1,2-dioxygenases and some other dioxygenases that catalyze the intradiol aromatic-ring-cleavage. Hybridization with the tfdC PCR product from the 2,4-D degradative plasmid pJP4 under stringent conditions revealed different extents of homology of the chlorocatechol 1,2-dioxygenase genes to the canonical tfdC sequence in the various strains. These findings were confirmed by the nucleotide sequence analysis of the tfdC-specific PCR products. From our results, we conclude that the PCR primer set is more suitable than the hybridization with pJP4-derived gene probes for the detection of diverse chlorocatechol 1,2-dioxygenase genes in proteobacteria.
    Additional Material: 3 Ill.
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  • 12
    Electronic Resource
    Electronic Resource
    Berlin : Wiley-Blackwell
    Acta Biotechnologica 18 (1998), S. 243-254 
    ISSN: 0138-4988
    Keywords: Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Process Engineering, Biotechnology, Nutrition Technology
    Notes: The degradation of lignocellulosic biomass of banana pseudoste was investigated during solid state fermentation (SSF) by P. ostreatus and P. sajor-caju. Both organisms proved to be efficient degraders of banana pseudostem biomass. P.ostreatus degraded hemicellulose (40% of dry weight, d.w.) better than cellulose (17.5% of d.w.) and lignin (10% of d.w.). P. sajor-caju also degraded hemicellulose (31% of d.w.) better than cellulose (12.4% of d.w.) and lignin (6% of d.w.). In both cases, a preferential removal of hemicellulose during the initial growth period and a delayed degradation of lignin were observed. The kinetics of cellulolytic, hemicellulolytic and lignolytic enzyme production in liquid culture were also examined. The activities of CMCase and β-glucosidase were highest at 16 days of growth and avicelase activity was at its maximum after 24 days (CMCase - 1.1 IU/ml, β-glucosidase - 0.09 IU/ml in the case of P. ostreatus; CMCase - 1.0 IU/ml, β-glucosidase - 0.087 - IU/ml in the case of P. sajor-caju.). Xylanase and laccase activity reached their maximum after day 16 and day 24 of incubation, respectively. (Xylanase - 1.1 IU/ml and laccase 3.0 IU/ml in the case of P. ostreatus; xylanase - 1.0 IU/ml and laccase - 3.6 IU/ml in the case of P. sajor-caju.). The efficient degrading capacity of test fungi demonstrated their potential use in the conversion of banana pseudostem biomass into mycelial protein-rich fermented animal feed.
    Additional Material: 4 Ill.
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  • 13
    Electronic Resource
    Electronic Resource
    Berlin : Wiley-Blackwell
    Acta Biotechnologica 18 (1998), S. 275-276 
    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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  • 14
    Electronic Resource
    Electronic Resource
    Berlin : Wiley-Blackwell
    Acta Biotechnologica 18 (1998), S. 305-313 
    ISSN: 0138-4988
    Keywords: Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Process Engineering, Biotechnology, Nutrition Technology
    Notes: For the purpose of forming cells possessing more than three nuclei and of determining the factors inducing multinucleation, cells of Saccharomyces cerevisiae were treated with 0, 0.3, 0.5, and 1.0% [w/v] colchicine solution, with and without shaking. When the cells were treated with 1.0% [w/v] colchicine solution, the number of cells containing two to eight nuclei was the largest. The multinucleate cells could grow on potato dextrose agar medium and their multinucleate nature did not disappear for at least three generations. This means that such cells are genetically stable. The proliferation rate of the multinucleate cells was not superior to that of the original strain. However, by monitoring the weight loss of the flask, it was possible to indirectly estimate the increase in the alcohol production of the multinucleate cell. It was concluded that the shaking treatment and higher colchicine concentrations contributed to multinucleation.
    Additional Material: 3 Ill.
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  • 15
    Electronic Resource
    Electronic Resource
    Berlin : Wiley-Blackwell
    Acta Biotechnologica 18 (1998), S. 315-324 
    ISSN: 0138-4988
    Keywords: Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Process Engineering, Biotechnology, Nutrition Technology
    Notes: Candida rugosa was cultivated in a mixed-solid substrate containing coconut oil cake (COC) and fine and coarse wheat bran (1:1:1) with an initial water activity (aw) of 0.92. The substrate was modified by adding a mineral solution (5%), corn steep liquor (6%), maltose (2%), peptone (3%), olive oil (10%), gum arabic (0.4%), different fatty acids (0.3%) and Tweens (0.5%). Fermentation in a column fermenter significantly improved the lipase yield to 118.2 Units per gram of dry fermented substrate [U/gds] at 72 h. This result was obtained 24 hours earlier than in our former studies (87.76 U/gds at 96 h) in COC, and the yield showed a 38% increase. Growth was measured indirectly by determining the glucosamine content in the cell wall of the yeast contained in the fermented matter, after its hydrolysis.
    Additional Material: 6 Ill.
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  • 16
    Electronic Resource
    Electronic Resource
    Berlin : Wiley-Blackwell
    Acta Biotechnologica 18 (1998), S. 53-62 
    ISSN: 0138-4988
    Keywords: Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Process Engineering, Biotechnology, Nutrition Technology
    Notes: The root nodules of Melilotus alba DESR., a fodder legume, contained high amounts of IAA. A tryptophan pool present in the nodule might serve as a source of IAA production. Presence of IAA oxidase and peroxidase in the nodules indicated the metabolism of IAA, at least in part, in the nodules. The Rhizobium species isolated from the root nodules produced a high amount of IAA (190 μg/ml) from L-tryptophan supplemented basal medium. IAA production and microbial growth were coincident. The production of IAA by the Rhizobium sp. was increased by 315% when the medium was supplemented with lactose (1%), NiCl2 (10 μg/ml), cetyl pyridinium chloride (0.5 μg/ml) and glutamic acid (0.4%), in addition to L-tryptophan (3 mg/ml). The possible role of the rhizobial production of IAA on the rhizobia-legume symbiosis is 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: In a prospective study, nuclear DNA was extracted from colorectal tumours and normal mucosa which had been fixed in buffered formalin and embedded into paraffin. DNA-extraction was performed using three different methods: a commercial kit which was not especially created for this use; a known fast procedure without DNA-cleaning steps; and a more conventional DNA-preparation protocol with DNA-cleaning. Using the polymerase chain reaction (PCR), DNA was amplified by being targeted onto two β-globin fragments with different lengths (536 bp and 989 bp) and (CA)n repeats localized on chromosome 5q (D5S346) and chromosome 17p (TP53CA) with a length of about 100 bp for detection of microsatellite instability. The success rate of microsatellite amplification was 100% with all methods. The 536 bp β-globin fragment could be amplified with a success rate ranging from 40% to 100%. The amplification of the 989 bp β-globin fragment was unsuccessful. Significant differences were observed between the three methods in the final DNA concentration and DNA yield. In microsatellite instability studies of paraffin-embedded tissues, the investigator can expect a high success rate of nearly 100% using any of the described methods.
    Additional Material: 1 Ill.
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  • 18
    Electronic Resource
    Electronic Resource
    Berlin : Wiley-Blackwell
    Acta Biotechnologica 18 (1998), S. 92-92 
    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
    Electronic Resource
    Electronic Resource
    Berlin : Wiley-Blackwell
    Acta Biotechnologica 18 (1998), S. 108-108 
    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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  • 20
    Electronic Resource
    Electronic Resource
    Berlin : Wiley-Blackwell
    Acta Biotechnologica 18 (1998), S. 95-106 
    ISSN: 0138-4988
    Keywords: Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Process Engineering, Biotechnology, Nutrition Technology
    Notes: An enzymatic process to decrease the phenolic content in canola meal was investigated. The new method was based on the addition of an enzyme preparation from the white-rot fungus Trametes versicolor to concentrated meal-buffer slurries. This approach eliminated the extraction of the valuable meal components such as proteins and carbohydrates. Two systems were considered: (i) slurries with canola meal concentrations higher than 33% [w/v]; (ii) slurries with canola meal concentrations equal to or less than 12.5% [w/v] with n-hexane as the main component of the continuous phase.The concentration of sinapic acid esters decreased by 99% after a 1.5, 2 and 3 hour long treatment of the meal with an initial moisture content of 75% at 90°C, 70°C and 50°C, respectively. The process was carried out at temperaturs as high as 110°C. Both the enzyme and the moisture concentrations influenced the enzymatic process and their action was coupled. The concentration of oxygen strongly affected the process.The enzymatic process was able to be carried out in the presence of hexane as the main component of the continuous phase. The optimum temperature for such a process was 30-40°C, At 30°C, after 1 h of treatment, the meal phenolic content was decreased by 97%. The water uptake by the meal was diminished in the presence of hexane.
    Additional Material: 6 Ill.
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  • 21
    ISSN: 0138-4988
    Keywords: Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Process Engineering, Biotechnology, Nutrition Technology
    Notes: TNM-FH Lepidopteran insect cell culture medium containing 10% fetal bovine serum (FBS), while allowing limited vegetative growth of Paenibacillus larvae (wild-type strain), the causative agent of American foulbrood, contained no viable vegetative cells upon subculture, nor were any heat resistant spores produced in this medium alone. However, TNM-FH medium cotaining embryonic or midgut cells from Trichoplusia ni, hemocytes from Estigmene acrea, ovarian and embryonic cells from Spodoptera frugiperda, embryonic cells from Plutella xylostella, Spodoptera exigua and Pseudaletia unipuncta or ovarian cells from Lymantria dispar, supported both heavy vegetative cell growth and moderate production of heat resistant spores. EX-CELL 405 serum-free insect cell culture medium alone appeared to contain the appropriate nutrients required for both vegetative growth and sporulation of P. larvae. However, in the presence of embryonic cells from T. ni, limited vegetative growth occurred and the P. larvae cells appeared to die off. This was confirmed by the fact that no colony growth occurred upon subculture, nor were any heat resistant spores detected. This was true also in the presence of fat body cells from T. ni, except that a limited number of spores (4,000/ml) were detected in the form of cology-forming units (CFU) on plates following heating to 80°C for 20 minutes. In a parallel study with a wild-type strain of Bacillus popilliae, vegetative cells grew only in TNM-FH medium in the presence of mid-gut BTI-Tn-MG and ovarian (Tn-368) cells of T. ni. No heat resistant spores, however, were detected in any of the cultures. When BTI-Tn-MG and Tn-368 cells were further challenged with four variant cultures of B. popilliae, vegetative growth and limited sporulation were achieved. The BTI-Tn-MG cell line in TNM-FH medium produced as many as 12,000 spores/ml after 21 days in culture.
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  • 22
    ISSN: 0138-4988
    Keywords: Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Process Engineering, Biotechnology, Nutrition Technology
    Notes: The conversion of starch from unhydrolyzed cassava flour to ethanol by a pure culture of Endomycopsis fibuligera and by a co-culture of this amylolytic yeast and the bacterium Zymomonas mobilis was studied.The best overall results were obtained using the mixed culture. After 96 h of fermentation of a medium containing 150 g/l initial cassava starch, an ethanol concentration of 31.4 g/l, a productivity of 0.33 g ethanol/l × h and a yield of 0.21 g ethanol/g initial starch were reached. The highest yield (0.37 g/g) was obtained after 48 h when using a medium containing 50 g/l initial starch.
    Additional Material: 2 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
    Electronic Resource
    Electronic Resource
    Berlin : Wiley-Blackwell
    Acta Biotechnologica 18 (1998) 
    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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  • 25
    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: No Abstracts.
    Type of Medium: Electronic Resource
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  • 27
    Electronic Resource
    Electronic Resource
    Berlin : Wiley-Blackwell
    Acta Biotechnologica 18 (1998), S. 17-27 
    ISSN: 0138-4988
    Keywords: Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Process Engineering, Biotechnology, Nutrition Technology
    Notes: Common theories of microbial growth and physiology are formulated exclusively in terms of the isolated microorganisms - especially bacteria. This is, however, an inadmissible simplification because it is obvious that the organization of microbial populations and colonies follows certain general rules.Bacterial colonies are able to generate complex interfacial growth patterns similar to those observed during diffusion-limited growth processes in non-living systems. One reason for these patterns is assumed to be the ability of many bacteria to swarm in an active manner on a substrate surface. Therefore the models of bacterial colony growth incorporate “random walkers”, which move actively in response to a gradient in the concentration of nutrients and communicate with each other by means of a chemotactic feedback.A selected number of yeasts were tested with regard to their colony growth patterns depending on the medium parameters such as nutrient concentration. Growth patterns similar to those which were described in literature for bacteria were also found in these experiments. It concerns in particular growth types like compact growth, fractal growth and dense-branching growth.This result allows a hypothesis to be formulated, that - especially in the case of fractal growth patterns - wandering of cells on a substrate surface may be induced by uncontrolled “swimming” on a thin water film caused by the metabolic activity (e.g. respiration) of the cells on the surface of the agar.Furthermore it was found that an interplay between changes in the individual morphology of yeast cells and the morphology transitions takes place. Such growth patterns are known for Candida sp. which are able to form pseudomycel and blastospores.
    Additional Material: 10 Ill.
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  • 28
    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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  • 29
    Electronic Resource
    Electronic Resource
    Berlin : Wiley-Blackwell
    Acta Biotechnologica 18 (1998), S. 43-50 
    ISSN: 0138-4988
    Keywords: Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Process Engineering, Biotechnology, Nutrition Technology
    Notes: Pioneering research efforts in the handling of municipal sewage in developing countries have involved the use of water hyacinth (Eichhornea crassipes) to purify sewage for possible re-use of the effluent water for domestic purposes. The ability of water hyacinth to remove pollution from raw sewage has been found to be impaired by sewage toxicity. Trials were therefore carried out to adapt water hyacinth to toxicity and thereby increase its ability to remove pollutants from raw sewage. The plants were adapted using an active bio-degrader consisting of Pseudomonas aeruginosa, Escherichia coli, Klebsiella ozaenae, Klebsiella edwardsiella and Baccillus subtilis. The adaptation progressed through 20, 40, 60 and 80% sewage dilution until plants capable of growth in 100% raw sewage were obtained. Plants were observed for morphological growth and at four weeks, samples were collected for tissue analysis.The plants progressively absorbed nutrients from sewage up to the fourth week, when signs of toxicity were obsereved through wilting, loss of turgidity and reduction in leaf number. However, plants that survived through a series of adaptations under various sewage dilutions exhibited luxuriant growth on raw sewage. In synergy with the active bio-degrader, the efficiency of the adapted water hyacinth to remove pollutants (nutrients) from raw sewage was enhanced by 93%.
    Additional Material: 4 Tab.
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  • 30
    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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  • 31
    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
    Electronic Resource
    Berlin : Wiley-Blackwell
    Acta Biotechnologica 18 (1998) 
    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
    Electronic Resource
    Electronic Resource
    Berlin : Wiley-Blackwell
    Acta Biotechnologica 18 (1998), S. 3-16 
    ISSN: 0138-4988
    Keywords: Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Process Engineering, Biotechnology, Nutrition Technology
    Notes: The reusability of biomass in lactic acid batch fermentation with free cells of Lactobacillus paracasei was studied in a 2-1 fermenter and in a 50-1 fermenter. In lab-scale fermentation experiments, 33 to 100% of the cell mass formed was reused in the subsequent batch in each case. In a series of seven consecutive batches, maximum values of lactate formation productivity of 6.32 to 11.54 g/l × h were observed at initial cell concentrations of 2.1 to 24.6 g/l. In all of the experiments, the initial cell viability was 78% or greater than 78%, and the final cell viability did not fall below 70%. At cell concentrations above 20 g/l, the productivity of lactic acid formation did not increase further, but remained constant. Because its level could be influenced by varying the proportions between the content of yeast extract, peptone and initial cell mass (1:1:2, 1:1:1 and 3.3.1) in the medium and no inhibitory effects were observed, this finding can be attributed to nutrient limitation. A low degree of cell reuse was reached in an analogous series of experiments carried out in a 50-1 fermenter. In this case, the initial cell concentration varied between 0.5 and 1.1 g/l, and therefore cell growth was not limited by nutrients in the first period of fermentation. Lactate production was still stable after six cell-reuse operations. The lactic acid yield did not fall below 90%. Temporary storage of the biomass in a refrigerator for a time interval of one to two weeks caused no significant impairment of overall lactate production, but a proportional prolongation of the lag phase occurred with increasing duration of storage.
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  • 34
    Electronic Resource
    Electronic Resource
    Berlin : Wiley-Blackwell
    Acta Biotechnologica 18 (1998), S. 29-41 
    ISSN: 0138-4988
    Keywords: Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Process Engineering, Biotechnology, Nutrition Technology
    Notes: A 4-nitrophenol degrading strain PNP1 isolated from the El-Harrach River near Algiers (Algeria) was studied with respect to its growth behaviour. According to the morphological and biochemical characteristics this strain was assigned to Pseudomonas putida. Besides 4-nitrophenol, the strain also used 1,2- and 1,4-dihydroxybenzene, benzoate, 4-hydroxybenzoate and 3,4-dihydroxybenzoate as sources of carbon and energy, degrading them exclusively via the ortho pathway. Pseudomonas putida PNP1 degrades 4-nitrophenol through a purely oxidative pathway with release of the nitro group as nitrite. During cultivation with 4-nitrophenol in ammonium-containing mineral medium, the strain PNP1 grew optimally at pH 7 and at a temperature between 30 and 35°C and showed stoichiometric nitrite release (at pH 7 and 30°C MONOD model parameters μmax = 0.615 h-1 and KS = 0.145 mg/l). A phenomenological model for the description of growth inhibition at high 4-nitrophenol concentrations was derived (below 400 mg/l only weak inhibition and at 600 mg/l acute toxicity). In ammonium-free medium, the maximum specific growth rate was reduced to 0.318 h-1 and part of the 4-nitrophenol-N was used as the nitrogen source (32% N in biomass and 68% N in nitrite). The yield coefficients of strain PNP1 were smaller in ammonium-free than in ammonium-containing medium (e.g. with 4-nitrophenol YX/S = 0.305 g/g compared to 0.350 g/g), which can be explained by the energy expense for the assimilatory nitrite reduction in the biosynthesis of N-containing cellular compounds. But the oxygen consumption was only slightly influenced by the ammonium content of the cultivation medium (e.g. with 4-nitrophenol YO/S = 1.005 g/g compared to 0.954 g/g).
    Additional Material: 5 Ill.
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  • 35
    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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  • 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.
    Type of Medium: Electronic Resource
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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.
    Type of Medium: Electronic Resource
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  • 38
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    Electronic Resource
    Berlin : Wiley-Blackwell
    Acta Biotechnologica 18 (1998), S. 63-75 
    ISSN: 0138-4988
    Keywords: Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Process Engineering, Biotechnology, Nutrition Technology
    Notes: An enzymic characteristic of Novo dextranase was presented. In addition to a high dextranolytic activity (7,200 U/ml), the crude enzyme also contained small amounts of protease, glucoamylase, polygalacturonase, carboxymethylcellulase, laminarinase and chitinase. A highly purified dextranase was then simply separated from a commercial preparation by column chromatographies on DEAE-Sepharose, CM-Sepharose, and by chromatofocussing on Polybuffer Exchanger PBE-94. The enzyme was recovered with an over 200-fold increase in specific activity and a yield of 84%. The final preparation was homogeneous, as observed during high performance liquid chromatography (HPLC). Size-exclusion HPLC indicated that dextranase had a molecular mass of 35 kDa and its isoelectric point, established by chromatofocussing, was 4.85. Analysis of the dextran break-down products indicated that purified dextranase represents an endolytic mode of action, and isomaltose and isomaltotriose were identified as the main reducing sugars of dextran hydrolysis. The enzyme was then covalently coupled to the silanized porous glass beads modified by glutaraldehyde (Carrier I) or carbodiimide (Carrier II). It was shown that immobilization of dextranase gave optimum pH and temperature ranges from 5.4 to 5.7 and from 50°C to 60°C, respectively. The affinity of the enzyme to the substrate decreased by a factor of more than 13 for dextranase immobilized on Carrier I and increased slightly (about 1.4-times) for the enzyme bound to Carrier II.
    Additional Material: 6 Ill.
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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: The ability of a mixed natural microbial population, collected in an aerated lagoon treating Fluff pulp effluent and Streptomyces viridosporus strain T7A, to degrade lignosulphonate was evaluated. S. viridosporus growing in a mineral medium containing glycerol (7 g/l) and lignosulphonate (1 g/l) allowed 20% of lignosulphonate to be degraded after 18 days of incubation. A culture of the mixed population on culture medium after S. viridosporus growth was unable to degrade lignosulphonate products. Moreover, antagonism between S. viridosporus and the mixed population or between S. viridosporus and the isolated strains from this population was observed. The enhancement of lignosulphonate biodegradation by naturally occurring microorganisms in association with S. viridosporus (bioaugmentation strategy) seems to be difficult.
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  • 40
    Electronic Resource
    Electronic Resource
    Berlin : Wiley-Blackwell
    Acta Biotechnologica 18 (1998) 
    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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  • 41
    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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  • 42
    Electronic Resource
    Electronic Resource
    Berlin : Wiley-Blackwell
    Acta Biotechnologica 18 (1998) 
    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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  • 43
    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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  • 44
    ISSN: 0138-4988
    Keywords: Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Process Engineering, Biotechnology, Nutrition Technology
    Notes: The cleaning of the exhaust gases of a bioreactor containing volatile hydrocarbons in a bioreactor system with a closed gas circuit is described. The bioreactor system consisted of three different reactor types: a stirred tank which was filled with hydrocarbon-containing waste water to simulate the exhaust gases of a remediation process; a trickle-bed reactor for aerobic treatment of the exhaust gas from the stirred tank; and a photoreactor containing an algae culture which assimilated CO2 from the trickle-bed reactor and also produced O2. With this bioreactor system, it was possible to efficiently remove volatile organic compounds (VOC) from the waste gases. Depending on the type of waste water investigated, elimination rates of 41% to 93% of BTEX (benzene, ethylbenzene, toluene, xylene) and 29% to 53% of VCH (volatile chlorinated hydrocarbons) were obtained. Due to the photosynthesis of the algae in the system's photoreactor, oxygen concentrations between 12% and 18% [v/v], equivalent to about 57% to 83% DOT, were obtained. This concentration permitted the aerobic degradation to be carried out without having to add fresh air. The trickle-bed reactor and the photoreactor worked continuously, whereas the waste water in the stirred bioreactor was replaced in different batches. The accumulation of toxic compounds in the nutrient solutions of the trickle-bed (EC-50 〉 30 g/l) and of the photoreactor (EC-50 〉 35 g/l) was low. Carbon dioxide concentrations in the gas flow were higher than in fresh air (1% to 3% [vol/vol]), but no long-term accumulation of CO2 occurred. This means that the algae in the photoreactor were active enough to assimilate the CO2 which had been produced. They were also able to produce sufficient oxygen for aerobic hydrocarbon degradation. The system described is a first step towards treating waste gases which results from the bioremediation of hydrocarbon-contaminated media in a closed gas circuit without any emission (e.g. VOC, CO2, germs).
    Additional Material: 3 Ill.
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  • 45
    Electronic Resource
    Electronic Resource
    Berlin : Wiley-Blackwell
    Acta Biotechnologica 18 (1998), S. 325-326 
    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
    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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  • 47
    Electronic Resource
    Electronic Resource
    Berlin : Wiley-Blackwell
    Acta Biotechnologica 18 (1998), S. 353-359 
    ISSN: 0138-4988
    Keywords: Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Process Engineering, Biotechnology, Nutrition Technology
    Notes: Protoplasts of a xylose-fermenting yeast strain (a fusion product of Pachysolen tannophilus and Saccharomyces cerevisiae) were fused with isolated nuclei of the xylan degrading filamentous fungus Fusarium moniliforme. Polyethyleneglycol 4000 was used as the fusogenic agent. Fourteen stable hybrids showing xylanase activity were obtained. It can be assumed that this ability was acquired from the nuclear genome of the fungus, since the parental yeast strain did not show any xylanase activity. The enzymatic activity was determined quantitatively. The parental strain of the fungus reached its maximum xylanase activity of 796 nkat/ml at 96 h of growth. Four of the hybrids had a xylanase activity of between 211 and 297 nkat/l at 24 h of growth. Zymograms of these hybrids showed the presence of xylanases when grown on xylan as the sole carbon source. Using pulse field electrophoresis gels, no difference between the chromosome pattern of the fusion products and the parental yeast strain was observed.
    Additional Material: 2 Ill.
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  • 48
    Electronic Resource
    Electronic Resource
    Berlin : Wiley-Blackwell
    Acta Biotechnologica 18 (1998), S. 367-367 
    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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  • 49
    Electronic Resource
    Electronic Resource
    Berlin : Wiley-Blackwell
    Acta Biotechnologica 18 (1998), S. 361-366 
    ISSN: 0138-4988
    Keywords: Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Process Engineering, Biotechnology, Nutrition Technology
    Notes: The calluses of two hydroxyproline-resistant lines (D20-1 and D30-1) of Solanum tuberosum L. were transferred to a solidified MS medium containing 1.0 mg/I IAA, 2.0 mg/l zeatin, 40.0 mg/l adenine sulphate, 1 g/l casein hydrolysate, 20 g/l sucrose and 10 g/l agar for plant regeneration. The shoot regeneration was only achieved from the callus of line D20-1. Regenerated shoots exhibited morphological variability. The degrees of frost tolerance were higher in the leaves of the regenerated plants compared with the leaves of the non-selected control plants, but lower than that of the callus from which they were regenerated.
    Additional Material: 1 Ill.
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  • 50
    Electronic Resource
    Electronic Resource
    Berlin : Wiley-Blackwell
    Acta Biotechnologica 18 (1998), S. 327-338 
    ISSN: 0138-4988
    Keywords: Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Process Engineering, Biotechnology, Nutrition Technology
    Notes: The batch production of gluconic acid in the presence of glucose, sucrose and molasses was investigated using free mycelia of Aspergillus foetidus NRRL 337 in shake flasks. Eight growth parameters were chosen as independent variables. The temperature, pH, substrate type and initial concentrations, inoculum percentage and shake rate directly affected the specific microorganism growth and gluconic acid production rates. The optimum temperature and initial pH values were found to be 33°C and five to six, respectively. The maximum specific growth and gluconic acid production rates were established as 57 g/dm3 of glucose, 75 g/dm3 of sucrose and 150 g/dm3 of molasses. The optimum values of the shake rate, inoculum percentage and initial ammonium nitrate concentration were determined as 100 1/min, 0.5% and 1.5 g/dm3, respectively. The maximum gluconic acid concentrations corresponding to these initial substrate concentrations were observed to be 8.3 g/dm3, 17.4 g/dm3 37.0 g/dm3, respectively. The optimum specific microbial growth and gluconic acid production rates were found as 0.0145 1/h and 0.0375 g/g × h, respectively, for the fermentation conditions of SGo = 57 g/dm3, T = 28°C, initial pH = 6.5, N = 84 1/min, A = 0.5 g/dm3 and I = 0.5%.
    Additional Material: 3 Ill.
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  • 51
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    Berlin : Wiley-Blackwell
    Acta Biotechnologica 18 (1998), S. 339-351 
    ISSN: 0138-4988
    Keywords: Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Process Engineering, Biotechnology, Nutrition Technology
    Notes: Covalent immobilization of thermostable α-amylases from catabolite resistant and sensitive Bacillus licheniformis strains on controlled pore glass (CPG) and porous silica (Spherosil) beads and ionic binding on DEAE-cellulose, Amberlite and Dowex were investigated. Preparations with satisfactory operational stabilities and activities up to 1,600 U/g of support (ionic binding) and 800 U/g carrier (covalent coupling) were obtained. Immobilization led to a narrowing of the pH interval of maximum activity. The fixed amylases were stable in limited pH regions around the optimum pH level. An enhancement of the enzyme thermostability was observed. Apparent shifts of the optimum temperatures were not found. The apparent Vmax decreased up to 80 times. The Km′ remained unchanged (for amylopectin as the substrate) or increased up to 10 times (soluble starch). Maltose, maltotriose and maltopentaose were the main products of the hydrolysis. A significant increase in maltopentaose content was observed.
    Additional Material: 3 Ill.
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  • 52
    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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  • 53
    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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  • 54
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    Electronic Resource
    Berlin : Wiley-Blackwell
    Acta Biotechnologica 18 (1998), S. 135-146 
    ISSN: 0138-4988
    Keywords: Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Process Engineering, Biotechnology, Nutrition Technology
    Notes: The formation of reactive groups on polyamide nets (nylon 6) and the subsequent immobilization of glucoamylase were investigated. Different mesh sizes of the nets and two chemical methods of enzyme coupling - i( partial hydrolysis of the polyamide with subsequent glutaraldehyde binding and ii) O-alkylation of the carrier using a treatment with a benzene-methyl sulphate mixture - were used. The reactivity of immobilized glucoamylase (GA) was tested by hydrolysis reactions using 1% starch solutions. The highest reactivity (140 μg glc/)min × cm2 was obtained for methylated nylon samples attached to a glass rod and by coupling glucoamylase on the nylon surface which had been treated with lysine and glutaraldehyde. This method resulted in a more reactive and more stable preparation of immobilized glucoamylase as compared to a simpler method of coupling glutaraldehyde to partially hydrolyzed nylon.
    Additional Material: 8 Ill.
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  • 55
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    Berlin : Wiley-Blackwell
    Acta Biotechnologica 18 (1998), S. 148-148 
    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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  • 56
    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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    Berlin : Wiley-Blackwell
    Acta Biotechnologica 18 (1998), S. 157-166 
    ISSN: 0138-4988
    Keywords: Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Process Engineering, Biotechnology, Nutrition Technology
    Notes: Different cultural parameters that regulate pectinolytic enzyme production in vitro by Trametes trogii were studied. When grown in a medium containing pectin, T. trogii produced extracellular polymethylgalacturonase, polygalacturonase and pectin lyase but no pectate lyase activity. No significant differences in the maximum enzyme activities measured were observed with the addition of xylan, carboxymethylcellulose or both to the medium containing pectin. The addition of glucose to that medium considerably decreases all the activities studied, and in a medium with glucose as the sole carbon source no galacturonase activity could be measured, and pectin lyase activity was at its minimum. The low synthesis of pectin lyase in cultures containing glucose suggests that this enzyme is constitutive in contrast to the polygalacturonases that were not detected. The increase in pectin concentration stimulated growth and enzyme production. The highest specific activities were attained with the greatest concentration tested (15 g/l). Casamino acids were the best nitrogen source for enzyme production. Maximum growth was measured at pH 3.3; pH values of around 4.5 stimulated enzyme production, but high pectinase activities were also detected in media with more alkaline initial pH values (6.2 for galacturonases and 6.6 for lyases), probably owing to the specific induction of particular isoforms. In the range of 23 to 28°C, good results were obtained in growth as well as in enzyme production. The addition of Tween 80 promoted growth and gave the highest yield of polymethylgalacturonase and pectin lyase (0.37 and 36.2 E.U./ml, respectively). The highest polygalacturonase activity (1.1 E.U/ml) was achieved with polyethylene glycol. Tween 20 and Triton X-100 inhibited growth and pectinase production.
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  • 58
    ISSN: 0138-4988
    Keywords: Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Process Engineering, Biotechnology, Nutrition Technology
    Notes: Cell growth and organic acid production by Propionibacteria are dependent on the vitamin-nitrogen source in the culture medium. Final cell and propionic acid concentrations produced by Propionibacterium shermanii, using corn-steep liquor, were higher than those obtained utilizing yeast extracts. Since corn-steep liquor is much cheaper than yeast extract, the process becomes more attractive. By calculating the specific growth rates, it was observed that the critical propionic acid concentration, that prevents all growth (μX = 0), is different depending on the vitamin-nitrogen source used and its concentration. For example, for 5.0 and 15.0 g/l Oxoid yeast extract, those critical propionic acid concentrations were 16.0 and 27.0 g/l, respectively. Such propionic acid concentrations inhibit the cell growth, but not the formation of acid. The specific propionic acid production rate also indicates that the critical concentration for metabolic activity, when propionic acid is no longer produced (μP = 0), varies according to the vitamin-nitrogen source and its concentration in the medium. For 5.0 and 15.0 g/l Oxoid yeast extract, those concentrations were 22.1 and 30.1 g/l, respectively.
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  • 59
    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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  • 60
    ISSN: 0138-4988
    Keywords: Life Sciences ; Life Sciences (general)
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Process Engineering, Biotechnology, Nutrition Technology
    Notes: The fermentation process of acid curd whey using pure cultures of L. bulgaricus and L. acidophilus was investigated. The influence of the starter culture amount on the acidification rate in the fermentation was specified, the biological value of fermented and fermented-ammoniated curd whey was determined, and the ability of fermented whey to prevent the injurious effect of Bac. mesenthericus on the wheat bread quality was examined.Acid curd whey was fermented up to a titratable acidity of 19.8-21.6 g lactic acid/kg whey using L. acidophylus and L. bulgaricus. Mathematical equations were developed on the basis of experimental data to calculate the titratable acidity (A) as a functionof fermentation time (τ) and temperature (t). Fermentation and fermentation-ammoniation processes increase the biological value of whey (the content of the vitamins B1, B2, B6, PP and the free amino acids increase). A new dry fodder BIOLAKTS was developed from fermented curd whey and was recommended for use in veterinary medicine. The fermentation-ammoniation process of curd whey was carried out by adding calculated amounts of non-protein nitrogen NH4OH to increase the total protein equivalent and to achieve mutual proportions of protein and lactose 1:1.4, as in skimmed milk. Fermented-ammoniated curd whey was used to obtain a skimmed milk substitute. A dry flour lactic acid concentrate (FLC) was created as a mixture of high quality wheat flour and evaporated fermented whey in established ratios. As our experiments prove, it can be used as an additive in bread-making to prevent the spoiling of wheat bread by Bac. mesenthericus.
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  • 61
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    Journal of Supramolecular Structure 3 (1975), S. 391-400 
    ISSN: 0091-7419
    Keywords: Life Sciences ; Molecular Cell Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Epithelial cells (80-90% “granular” type) were isolated from urinary bladders of Bufo marinus and Rana catesbiana. The inhibitory effect of α-methyl-D-mannoside on fluorescein-labeled concanavalin A (Con A) binding to these cells indicates that they possess specific binding sites for Con A. The lectin also mediates adsorption of erythrocytes to these cells. Both Con A binding and Con A-mediated hem-adsorption to epithelial cells are depressed at 4°C, as compared with cells maintained at 22°C. Elevation of temperature to 37°C, however, enhances hemadsorption independently of alterations in lectin binding. Treatment of cells with antidiuretic hormone (ADH) at 22°C followed by 15 min of incubation at 22° or 37°C before exposure of cells to Con A promotes increments in Con A-mediated hemadsorption, but not in lectin binding, at 22° or 37°C. These hormonal effects are not significant when hemadsorption is assayed at 4°C. Treatment of cells with another octapeptide, angiotensin, elicits a small, but significant, increment in hemadsorption to epithelial cells which is likewise uninfluenced by quantitative changes in lectin binding. Collectively, these data and other independent observations suggest that treatment with octapeptide hormones acts to enhance the redistribution and aggregation of lectin-binding proteins in the membranes of granular epithelial cells from amphibian urinary bladder. Such changes, in turn, may contribute to the alterations in membrane transport properties which characterize the hormonal response.
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  • 62
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    Journal of Supramolecular Structure 3 (1975), S. 376-390 
    ISSN: 0091-7419
    Keywords: Life Sciences ; Molecular Cell Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: According to current models of muscle contraction (Huxley, H. E., Science 164: 1356-1366 [19691]), motion of flexible myosin crossbridges is essential t o the contractile cycle. Using a spin-label analog of iodoacetamide bound to the subfragment # 1 (S1) region of myosin, we have obtained rotational correlation times (τ2) for this region of the molecule with the ultimate goal of making quantitative measurements of the motion of the crossbridges under conditions comparable to those in living, contracting muscle. We used the newly developed technique of saturation transfer electron paramagnetic resonance spectroscopy (Hyde, J.S., and Thomas, D.D., Ann. N.Y. Acad. Sci. 222:680-692 [1973]), which is uniquely sensitive t o rotational motion in the range of 10-7-10-3 sec. Our results indicate that the spin label is rigidly bound to S1 (τ2 for isolated S1 is 2 × 10-7 sec) and that the motion of the label reflects the motion of the S1 region of myosin. The value of τ2 for the S1 segment of myosin is less than twice that for isolated S1, while the molecular weights differ by a factor of 4, indicating flexibility of myosin in agreement with the conclusions of Mendelson et aL (Biochemistry 12:2250-2255 [1973]). Adding F-actin increases τ2 in either myosin or isolated S1 by a factor of nearly 103, indicating rigid immobilization of S1 by actin. Formation of myosin filaments (at an ionic strength of 0.15 or less) increases τ2 by a factor of 10-30, depending o n the ionic strength, indicating a decrease of the rotational mobility of S1 in these aggregates. The remaining motion is at least a factor of 10 faster than would be expected for the filament itself, suggesting motion of the S1 region independent of the filament backbone but slower than in a single molecule. F-actin has a strong immobilizing effect on labeled S l in myosin filaments (in 0.137 M KC1), but the immobilization is less complete than that observed when F-actin is added t o labeled myosin monomers (in 0.5 M KC1). A spin-label analog of maleimide, attached to the SH-2 thiol groups of S1, is immobilized to a much lesser extent by F-actin than is the label on SH-1 groups. The maleimide label also was attached directly to F-actin and was sufficiently immobilized to suggest rigid binding to actin.
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  • 63
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    Journal of Supramolecular Structure 3 (1975), S. 112-124 
    ISSN: 0091-7419
    Keywords: Life Sciences ; Molecular Cell Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Evidence for a proposed activation mechanism is summarized. The low rate of ATP cleavage in the resting state of muscle is considered to result from the formation of a stable ring structure involving the two essential sulfhydryl groups on each myosin head and MgATP. Activation is thought to occur by interaction of actin in the vicinity of one of the essential sulfhydryl groups, Thus opening the stable ring leading to rapid dissociation of split products. This idea is consistent with the kinetic scheme of ATP cleavage developed recently by other workers and allows a prediction of the shift in population of intermediate states with changes in solvent conditions. It is also supported by our recent studies on the spatial geometry of the ring. The possibility that other nucleophilic groups may replace the sulfhydryl groups in other contractile systems is considered. The relevance of the ring structure to the tension generating event is discussed on the basis of recent measurements of the rate of contraction of modified (SH1-blocked) actomyosin threads. Results indicate that the ability to form the ring structure is an essential requirement of the contractile process in these systems, and, moreover, that single, modified heads of myosin can act independently to produce the same rate of contraction as native myosin. This latter finding suggests that the myosin duplex exhibits some type of negative cooperativity in the contractile process.
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  • 64
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    Journal of Supramolecular Structure 3 (1975), S. 146-153 
    ISSN: 0091-7419
    Keywords: Life Sciences ; Molecular Cell Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: The extent of actin polymerization has been studied for samples in which the bound nucleotide of the actin was ATP, ADP, or an analog of ATP that was not split (AMPPNP). The equilibrium constants for the addition of a monomer to a polymer end were determined from the concentration of monomer coexisting with the polymer. An analysis of these results concludes that the bound ATP on G-actin provides little energy to promote the polymerization of the actin. AMPPNP was incorporated into F-actin and the interaction of F-actin · AMPPNP with myosin was studied. F-actin · AMPPNP activated the ATPase of myosin to the same extent as did F-actin · ADP. However, the rate of superprecipitation was slower in the case of F-actin · AMPPNP than in the control.
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  • 65
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    Journal of Supramolecular Structure 3 (1975), S. 256-260 
    ISSN: 0091-7419
    Keywords: Life Sciences ; Molecular Cell Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: 1,N6-Ethenoadenosine diphosphate (∊-ADP) inhibits reverse electron flow (succinate → NAD+ driven by ATP) by competing with ATP, in contrast to ADP which we have shown previously to be a noncompetitive inhibitor. From these and other data it is concluded that the noncompetitive inhibition noted with ADP results from a combination of competitive inhibition plus non- or uncompetitive inhibition, the former occuring at a relatively nonspecific catalytic site and the latter at an extracatalytic site apparently quite specific for ADP.ADP, which stimulates ATP ⇌ H2O and Pi ⇌ H2O exchanges appears to be necessary for inhibition by arsenate of these exchanges. It is suggested that the ATP-supported Pi ⇌ H2O exchange may be predominantly of the medium or intermediate type, depending on the concentrations of the Mg2+ complexes of ADP and Pi. Thus only exchanges involving medium ADP and Pi would be expected to show arsenate sensitivity.
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  • 66
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    Journal of Supramolecular Structure 3 (1975), S. 261-274 
    ISSN: 0091-7419
    Keywords: Life Sciences ; Molecular Cell Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Highly purified preparations of Streptococcus faecalis ATPase contain a similar but inactive protein detected by prolonged polyacrylamide gel electrophoresis. The inactive protein appears to arise by proteolytic cleavage of the major subunits in the enzyme. By use of a new technique, subunit analysis in SDS gels was performed on the enzyme band and the inactive protein band excised from a polyacrylamide gel after electrophoresis. The results indicated that the ATPase has the composition α3β3γ in which α = 60,000, β = 55,000, and γ = 37,000 daltons. The inactive protein appears to have the composition (f)6 in which f = 49,000 daltons. There is also evidence that the enzyme band contains some slightly modified forms of the ATPase, such as α3β2 (f)γ. The inactive protein lacks the capacity for tight nucleotide binding.Our experiments show that the tight ATPase-nucleotide complex formed in S. faecalis cells (the endogenous complex) behaves differently from the tight complex formed in vitro (the exogenous complex). We prepared a doubly labeled complex containing endogenous 32P-labeled ADP and ATP and exogenous 3H-labeled ADP. We observed that the addition of free nucelotide to the doubly labeled ATPase displaced the exogenous bound ligand from the enzyme but not the endogenous bound nucleotide. We suggest that the displaceable and nondisplaceable forms of the tight ATPase-nucleotide complex correspond to two different conformational states of the enzyme.
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  • 67
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    Journal of Supramolecular Structure 3 (1975) 
    ISSN: 0091-7419
    Keywords: Life Sciences ; Molecular Cell Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
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  • 68
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    Journal of Supramolecular Structure 3 (1975), S. 1-16 
    ISSN: 0091-7419
    Keywords: Life Sciences ; Molecular Cell Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: The choline-containing teichoic acids of pneumococci can be modified by biosynthetic replacement of the choline residues with certain structural analogues, such as ethanolamine (EA) or the N-monomethyl- (MEA) and N-dimethyl- (DEA) amino derivatives of ethanolamine. Cells containing such analogues in their teichoic acids develop pleiomorphic alterations in several physiological properties, which include resistance to detergent-induced lysis and inhibition of cell separation (chain formation). We report here the results of physiological studies on the mechanism of these two phenomena. Our results are summarized in the following: (a) Pneumococci grown on various amino alcohols produce cell walls of identical amino sugar and amino acid composition. (b) Both choline- and EA-containing teichoic acids seem to follow the same conservative pattern of segregation during growth and cell division. (c) Lysis sensitivity of pneumococci requires the juxtaposition of lysissensitive (choline-containing) cell walls and endogenous autolysin at the cell wall growth zone. (d) Upon readdition of choline to ethanolamine-containing cells, lysis sensitivity and catalytically active (C-type) autolysin reappear in the bacteria with the same kinetics. (e) The chains of EA-grown pneumococci contain fully compartmentalized cells and normal cross walls.
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  • 69
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    Journal of Supramolecular Structure 3 (1975), S. 24-38 
    ISSN: 0091-7419
    Keywords: Life Sciences ; Molecular Cell Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: The protein products of at least 21 phage genes are needed for the formation of the tail of bacteriophage T4. Cells infected with amber mutants defective in these genes are blocked in the assembly process. By characterizing the intermediate structures and unassembled proteins accumulating in mutant-infected cells, we have been able to delineate most of the gene-controlled steps in tail assembly. Both the organized structures and unassembled proteins serve as precursors for in vitro tail assembly.We review here studies on the initiation, polymerization, and termination of the tail tube and contractile sheath and the genetic control of these processes. These studies make clear the importance of the baseplate; if baseplate formation is blocked (by mutation) the tube and sheath subunits remain essentially unaggregated, in the form of soluble subunits.Seventeen of the 21 tail genes specify proteins involved in baseplate assembly. The genes map contiguously in two separate clusters, one of nine genes and the other of eight genes. Recent studies show that the hexagonal baseplate is the end-product of two independent subassembly pathways. The proteins of the first gene cluster interact to form a structure which probably represents one-sixth of the outer radius. The products of the other gene cluster interact to form the central part of the baseplate.Most of the phage tail precursor proteins appear to be synthesized in a non-aggregating form; they are converted to a reactive form upon incorporation into preformed substrate complexes, without proteolytic cleavage. Thus reactive sites are limited to growing structures.
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  • 70
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    Journal of Supramolecular Structure 3 (1975), S. 51-60 
    ISSN: 0091-7419
    Keywords: Life Sciences ; Molecular Cell Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Lactoperoxidase-catalyzed iodination selectively labels the two glycoproteins (VP1 and VP2) of Newcastle disease virus. The low-molecular-weight, nonglycosylated major viral protein, VP6, was not iodinated in the intact virus but was iodinated in disrupted virions, suggesting a localization on the inner, rather than the outer envelope surface. Studies on the distribution of virion proteins labeled with 125I and 3H-isoleucine between detergent-soluble and detergent-insoluble fractions show that the virion proteins VP4, VP5, and VP6 are solubilized to a much lesser extent than are VP1 and VP2.
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  • 71
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    Journal of Supramolecular Structure 3 (1975) 
    ISSN: 0091-7419
    Keywords: Life Sciences ; Molecular Cell Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
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  • 72
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    Journal of Supramolecular Structure 3 (1975), S. 105-111 
    ISSN: 0091-7419
    Keywords: Life Sciences ; Molecular Cell Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
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  • 73
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    Journal of Supramolecular Structure 3 (1975), S. 154-161 
    ISSN: 0091-7419
    Keywords: Life Sciences ; Molecular Cell Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: A new approach to the direct estimation of the value of the off constant for dissociation of ATP from myosin subfragment 1 (S1) has been developed. From measurements of the extremely slow rate of release of [32P]-ATP formed from 32Pi by S1 catalysis and the amount of rapidly formed [32P]-ATP tightly bound to S1, the value of the off constant is approximately 2.8 × 10-4 sec-1 at pH 7.4.The concentration dependencies for Pi ⇌ H18 OH exchange and for 32Pi incorporation into myosin-bound ATP give direct measurements of the dissociation constant of Pi from S1. Both approaches show that the enzyme has a very low affinity for Pi, with an apparent Kd of 〉 400 mM.Measurement of the average number of water oxygens incorporated into Pi released from ATP by S1-catalyzed hydrolysis in the presence of Mg2+ suggests that the hydrolytic step reverses an average of at least 5.5 times for each ATP cleaved. With the Ca2+-activated hydrolysis, less than one oxygen from water appears in each Pi released. This finding is indicative of a possible isotope effect in the attack of water on the terminal phosphoryl group of ATP.
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  • 74
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    Journal of Supramolecular Structure 3 (1975), S. 175-180 
    ISSN: 0091-7419
    Keywords: Life Sciences ; Molecular Cell Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Methods are described whereby initial processes of muscular contraction may be investigated in a mammalian preparation, the soleus muscle of the rat. Conditions are chosen so that recovery is avoided. An isometric tetanus is investigated and an energy balance sheet is drawn up. It is found that there is more heat evolved than can be accounted for in terms of measured chemical reaction. This discrepancy is discussed with reference to the similar results that have been obtained using frog muscle.
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  • 75
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    Journal of Supramolecular Structure 3 (1975), S. 169-174 
    ISSN: 0091-7419
    Keywords: Life Sciences ; Molecular Cell Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: From comparative studies of the association with polymeric actin of the bifunctional species heavy meromyosin and its monofunctional constituents, information about the relative freedom of these paired elements can be derived. An isotherm for the former binding process is presented which involves, as an experimentally determinable parameter, the local concentration of a second segment after the first of a pair is attached to the lattice. From combined data for these two association reactions a value of 10-4 M is obtained for this quantity. The large degree of segmental flexibility reported for the free heavy meromyosin is still manifested in the association with actin.
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  • 76
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    Journal of Supramolecular Structure 3 (1975), S. 162-168 
    ISSN: 0091-7419
    Keywords: Life Sciences ; Molecular Cell Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Time-dependent fluorescence depolarization and lifetime studies have been made on myosin subfragment 1 to obtain information about mobility changes and dye environment changes when different nucleotides are added. Data are reported for static and actively hydrolyzing systems containing G- and F-actin. Preliminary data indicate that myosin labeled with the fluorophore 1,5 IAEDANS1-and treated with DTT preserves its actin-activated Vmax. S1 prepared in this manner gives lifetime changes which are nearly identical for all systems studied. S1 labeling without DTT addition gives a pattern of lifetimes similar, though not identical to ESR work. Either type of labeling produces no observable change in the polarization decay, and we set an upper limit of 15% length change for the elongate S1. An unusually long fluorescence decay lifetime for the S1-Mg++ ATP-G-actin system is found which may indicate a new acto-S1 state stabilized by G-actin. The method for obtaining the bound fraction of S1's in the presence of actin is presented and applied to the S1-F-actin-Mg++ ATP system. Qualitative agreement is obtained with other methods.
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  • 77
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    Journal of Supramolecular Structure 3 (1975), S. 17-23 
    ISSN: 0091-7419
    Keywords: Life Sciences ; Molecular Cell Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: The temperature-dependent dissociation of neutral salt-soluble collagen into its component chains was measured in 0.6-1.6 M urea solutions at pH 7.3. The temperature-dependent association of the same radiocactively labeled collagen into fibers was measured in 0-0.4 M urea solutions, pH 7.3. The effect of urea on the temperature, Tm(G), for half dissociation into chains was small, and the value extrapolated to zero urea concentration was 39°C. In contrast, the effect of urea on the temperature, Tm(F), for half association into fibers was large, and the value at zero urea concentration was 30°C.We conclude that while body temperature provides excellent conditions for the matching of collagen chains to form molecules, the conditions are not optimal for the formation of highly ordered fibers. The large effects of 0.1 M urea suggest that other factors in vivo may help to destabilize mismatched molecular association during fiber growth. Alternately this might be facilitated by parts of the extension peptides of procollagen.
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  • 78
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    Journal of Supramolecular Structure 3 (1975), S. 39-50 
    ISSN: 0091-7419
    Keywords: Life Sciences ; Molecular Cell Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: The porcine neurotubule and its basic subunit were found to be modified in vitro by iodination of amino acids (principally tyrosine) using lactoperoxidase. Iodide ion, H2O2, or lactoperoxidase singly or in any pairwise combination had virtually no effect on neurotubules. However, when all three reagents were present, permitting covalent iodination, it was found that at 0.1 iodotyrosines per tubulin dimer the microtubules unravel to form structures which morphologically resemble strands of protofilaments twisted or wound around each other. These abnormal tubules are stable at room temperature and 4°C. Both monomers of tubulin are labeled to approximately the same extent. Iodinated tubulin (0.1 iodotyrosines/dimer) is unable to assemble in vitro under normal assembly conditions. Heavily iodinated microtubules (8 iodines per tubulin dimer) are similar in morphology to the slightly iodinated structures.
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  • 79
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    Journal of Supramolecular Structure 3 (1975), S. 61-66 
    ISSN: 0091-7419
    Keywords: Life Sciences ; Molecular Cell Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: To reveal the presence of rhodopsin on the surface of the mouse retina, a scanning electron microscope study of the immunolabeling of rhodopsin was attempted. The glutaraldehyde-fixed mouse retina was treated first with rabbit antibodies specific against bovine rhodopsin and then with hemocynin-labeled goat antibodies specific against rabbit antibody. The distribution of hemocyanin label on mouse retina and the technique used for labeling are discussed.
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  • 80
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    Journal of Supramolecular Structure 3 (1975), S. 125-140 
    ISSN: 0091-7419
    Keywords: Life Sciences ; Molecular Cell Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: The reactivity of myosin to [14C]-labeled N-ethylmaleimide ([14C] NEM) or to tritium was determined in functionally different frog muscles. The incorporation of [14C] NEM into myosin decreased during isotonic or isometric contractions, as compared to resting muscle. The cysteine residues which were protected during contraction were not involved in the ATPase activity or the actin-binding ability of myosin. Peptide mapping revealed that several residues were protected simultaneously. The incorporation of tritium into the peptide N-H groups of myosin was also decreased during muscle activity. These data support the idea that activation and subsequent contraction of muscle are correlated with structural changes in the myosin molecule.The reactivity of myosin to [14C] NEM was increased when the muscle was stretched to 140% rest length and treated with iodoacetate to deplete ATP. Based on in vitro experiments and on literature data, it is suggested that in the resting muscle myosin contains bound MgATP which decreases the rate of incorporation of [14C] NEM into myosin and that upon the irreversible loss of ATP the rate increases.31P nuclear magnetic resonance signals from a number of phosphates were detected in the intact frog muscle. The data indicated that the minimum concentration of ATP in the muscle is 3 mM, a value which agrees with that of chemical determination. The characteristic chemical shifts, coupling constants, and line widths of ATP in the muscle were considerably altered from that of either free ATP in aqueous solutions or ATP in perchloric acid extracts of muscle.
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  • 81
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    Keywords: Life Sciences ; Molecular Cell Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Recent reports in the literature have indicated a physical association of creatinephosphokinase (CPK) with the tail portion of the myosin molecule. The present paper describes further studies on the interaction of CPK with myosin and myosin fragments, using the techniques of electron paramagnetic resonance (EPR) and nanosecond fluorescence depolarization. From EPR work, spin-labeled CPK appears to interact with myosin, tail-less myosin (heavy meromyosin [HMM]), and myosin heads (subfragment-1 [S1]), the extent of interaction being proportional to the S1 content of myosin or its fragments. Spin-labeled CPK did not evidence interaction with the headless myosin “rods”, with myosin tails (light meromyosin [LMM]), with S2 necks (which connect S1 to the rest of the myosin molecule), or with actin. When a fluorescent dye is directed to the essential ∊-amino group of CPK, nanosecond fluorescence depolarization studies indicate a substantial interaction with myosin, HMM, and S1, but very little with F-actin. When the “fast-reacting” thiol of the S1 moiety or the “essential thiol” of CPK was labeled with either a fluorescent dye or a spin label, no interaction between CPK and myosin (or S1) was detected.
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  • 82
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    Keywords: Life Sciences ; Molecular Cell Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
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  • 83
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    Journal of Supramolecular Structure 3 (1975), S. 510-519 
    ISSN: 0091-7419
    Keywords: Life Sciences ; Molecular Cell Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: The effect of substrate on the far UV (185-250 nm) and near UV (250-325 nm) circular dichroism (CD) of myosin and heavy meromyosin (HMM) was studied. The following results were obtained with the addition of ATP (during various conditions of hydrolysis), ADP, and pyrophosphate: (1) no changes were observed in the far UV CD, (2) ATP and ADP perturbed the near UV CD only at spectral regions below 280 nm coinciding with the regions of their optical activity, (3) the optically inactive pyrophosphate caused no change in the near UV CD, and (4) myosin and HMM gave exactly the same results. These results suggest that myosin-substrate interaction in vitro does not result in a delocalized conformational change.
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  • 84
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    Journal of Supramolecular Structure 3 (1975), S. 80-89 
    ISSN: 0091-7419
    Keywords: Life Sciences ; Molecular Cell Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Fluorescent polymers were obtained by oxidizing partly emulsified linolenic acid with different oxidants. The speed of formation of polymers differed for the various oxidants, and the difference was not a simple function of the oxidation potential. The speed of polymerization also depended on the nature of the emulsion.The presence of egg albumen in the emulsion enhanced polymer formation with all oxidants. When the oxidants used are arranged in the order of decreasing speed of polymer formation, the order is different in the presence of albumen from what it is in the absence of albumen.With different oxidation catalysts most antioxidants and amino acids tested enhanced polymerization. In oxidation with ferric ions, with K-dichromate, and without added oxidants the only antioxidants which delayed polymerization were “inhibitors”. “Retarders” enhanced polymerization. With KMnO4 slight delay was caused by some retarders.The findings indicate that not only oxidation catalysts, but also proteins, amino acids, and antioxidants enhance polymerization. The possibility is suggested that in animal cells lipid pigment formation might represent a mechanism for neutralizing free radicals.
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  • 85
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    Journal of Supramolecular Structure 3 (1975), S. 304-313 
    ISSN: 0091-7419
    Keywords: Life Sciences ; Molecular Cell Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: EPR and water proton relaxation rate (1/T1) studies of partially (40%) and “fully” (90%) purified preparations of membrane-bound (Na++K+) activated ATPase from sheep kidney indicate one tight binding site for Mn2+ per enzyme dimer, with a dissociation constant (KD = 0.88 μM) in agreement with the kinetically determined activator constant, identifying this Mn2+-binding site as the active site of the ATPase. Competition studies indicate that Mg2+ binds at this site with a dissociation constant of 1 mM in agreement with its activator constant.Inorganic phosphate and methylphosphonate bind to the enzyme-Mn2+ complex with similar high affinities and decrease l/T1 of water protons due t o a decrease from four to three in the number of rapidly exchanging water protons in the coordination sphere of enzyme-bound Mn2+. The relative effectiveness of Na+ and K+ in facilitating ternary complex formation with HPO2-4 and CH3PO2-3 as a function of pH indicates that Na+ induces the phosphate monoanion t o interact with enzyme-bound Mn2+, while K+ causes the phosphate dianion to interact with the enzyme-bound Mn2+. Thus protonation of an enzyme-bound phosphoryl group would convert a K+-binding site to a Na+-binding site. Dissociation constants for K+ and Na+, estimated from NMR titrations, agreed with kinetically determined activator constants of these ions consistent with binding t o the active site.Parallel 32Pi-binding studies show negligible formation (〈 7%) of a covalent E-P complex under these conditions, indicating that the NMR method has detected an additional noncovalent intermediate in ion transport. Ouabain, which increases the extent of phosphorylation of the enzyme to 24% at pH 7.5 and t o 106% at pH 6.1, produced further decreases in l/T 1 of water protons. Preliminary 31P-relaxation studies of CH3PO2-3 in the presence of ATPase and Mn2+ yield an Mn to P distance (6.9 ± 0.5 Å) suggesting a second sphere enzyme-Mn-ligand-CH3PO2-3 complex.Previous kinetic studies have shown that T1+ substitutes for K+ in the activation of the enzyme but competes with Na+ at higher levels. From the paramagnetic effect of Mn2+ at the active site on the enzyme on I/T1 of 205T1 bound at the Na+ site, a Mn2+ to T1+ distance of 4.0 ± 0.1 Å is calculated, suggesting the sharing of a common ligand atom by Mn2+ and T1+ on the ATPase. Addition of P. increases this distance to 5.4 Å consistent with the insertion of P between Mn2+ and T1+. These results are consistent with a mechanism for the \documentclass{article}\pagestyle{empty}\begin{document}$ (\mathop {\rm N}\limits^{\rm i} {\rm a}^{\rm + } {\rm + K}^ +) $\end{document}-ATPase and for ion transport in which the ionization state of Pi at a single enzyme active site controls the binding and transport of Na+ and K+, and indicate that the transport site for monovalent cations is very near the catalytic site of the ATTase. Our mechanism also accounts for the order of magnitude weaker binding of Na+ compared to K+.
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  • 86
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    Journal of Supramolecular Structure 3 (1975), S. 103-103 
    ISSN: 0091-7419
    Keywords: Life Sciences ; Molecular Cell Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
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  • 87
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    Journal of Supramolecular Structure 3 (1975), S. 521-521 
    ISSN: 0091-7419
    Keywords: Life Sciences ; Molecular Cell Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Additional Material: 1 Ill.
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  • 88
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    Journal of Supramolecular Structure 3 (1975), S. 248-255 
    ISSN: 0091-7419
    Keywords: Life Sciences ; Molecular Cell Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: We have partially purified active delta and epsilon subunits of the E. coli membranebound Mg2+ -ATPase (ECF1). Treating purified ECF1 with 50% pyridine precipitates the major subunits (α, β, and γ) of the enzyme, but the two minor subunits (δ and ∊), which are present in relatively small amounts, remain in solution. The delta and epsilon subunits were then resolved from one another by anion exchange chromatography. The partially purified epsilon strongly inhibits the hydrolytic activity of ECF1. The epsilon fraction inhibits both the highly purified five-subunit ATPase and the enzyme deficient in the δ subunit. The latter result indicates that the delta subunit is not required for inhibition by epsilon. By contrast, two-subunit enzyme, consisting chiefly of the α and β subunits, was insensitive to the ATPase inhibitor, suggesting that the γ subunit may be required for inhibition by epsilon.The partially purified delta subunit restored the capacity of ATPase deficient in delta to recombine with ATPase-depleted membranes and to reconstitute ATP-dependent transhydrogenase. Previously we reported (Biochem. Biophys. Res. Commun. 62:764 [1975]) that a fraction containing both the delta and epsilon subunits of ECF1 restored the capacity of ATPase missing delta to recombine with depleted membranes and to function as a coupling factor in oxidative phosphorylation and for the energized transhydrogenase. These reconstitution experiments using isolated subunits provide rather substantial evidence that the delta subunit is essential for attaching the ATPase to the membrane and that the epsilon subunit has a regulatory function as an inhibitor of the ATPase activity of ECF1.
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  • 89
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    Journal of Supramolecular Structure 3 (1975), S. 333-337 
    ISSN: 0091-7419
    Keywords: Life Sciences ; Molecular Cell Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Myosin catalyzed exchange between 32Pi and ATP in reaction medium during its enzymatic hydrolysis of ATP only by a very small amount. Addition of actin increased to a great extent the rate of incorporation of 32Pi in the presence of Mg. Glycerinated smooth muscle fibers also exhibited the ability to exchange 32Pi and ATP upon the application of external force (repeated stretching and releasing). A schematic mechanism of the action of actin and external force on acceleration of 32Pi incorporation is proposed and the importance of the M*-ADP complex for force generation is suggested.
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  • 90
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    Journal of Supramolecular Structure 3 (1975), S. 315-322 
    ISSN: 0091-7419
    Keywords: Life Sciences ; Molecular Cell Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: From transient kinetic studies of the Mg2+-dependent adenosine triphosphatase of myosin subfragment 1, prepared from rabbit skeletal muscle, a seven-step mechanism has been proposed. Features of this mechanism include two-step processes for ATP and ADP binding in which the binary complex isomerizes in addition to a rapid nucleotide association step. In the case of ATP a large negative standard free energy change is associated with the isomerization. An overall rate-limiting isomerization of the myosin-product complex prior to product release has been identified. Studies on the mechanism of cleavage of ATP bound to the active site indicate the process is readily reversible and can account for the observation that more than one oxygen of the product phosphate arises from water. This proposal has been substantiated by the finding that the oxygen atoms of the γ-phosphoryl group of bound ATP also undergo extensive exchange with water.
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  • 91
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    Journal of Supramolecular Structure 3 (1975), S. 361-367 
    ISSN: 0091-7419
    Keywords: Life Sciences ; Molecular Cell Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: The binding of ADP to heavy meromyosin has been studied by microcalorimetry. Minute amounts of myokinase interfere with binding measurements, but by selection of appropriate conditions, we can estimate that the value of the apparent ΔHbinding lies between -1.0 and -3.0 kcal per mole of ADP bound (0.3 M KCl, 2 mM MgCl2, 20 mM Tris, pH 8.00, 20°C). Values of ΔHbinding reported to date are an order of magnitude larger, and we suggest that these values are artifactual results due to myokinase contamination.
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  • 92
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    Journal of Supramolecular Structure 3 (1975), S. 348-353 
    ISSN: 0091-7419
    Keywords: Life Sciences ; Molecular Cell Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Structural changes in myosin may take place within the heavy chains, or may represent rearrangement of subunits. Changes induced by ATP and possibly restricted to the heavy chains are indicated by several different optical studies, including changes in absorbence of aromatic residues (1), changes in the fluorescene of dyes coupled to myosin (2), changes in electron spin resonance spectrum (3), and changes in the intrinsic fluorescence of tryptophan residues (4, 5). A calcium dependent subunit rearrangement in scallop myosin is indicated by fluorescene depolarization studies (Cheung and Szent-Györgyi, unpublished observations). In scallop muscles, calcium regulates contraction by directly reacting with myosin (6), and the changes in the quaternary structure of myosin are likely to be related in the way calcium control works.At first I will disucss the changes of fluorenscene that Drs. Weber, Fasman, and I have described several years ago (4). Then I will speculate on the implications of a subunit rearrangement on the mechanism of regulatory light chain funciton.
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  • 93
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    Journal of Supramolecular Structure 3 (1975), S. 354-360 
    ISSN: 0091-7419
    Keywords: Life Sciences ; Molecular Cell Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: The interaction of myosin and paramyosin was investigated by enzymological and ultrastructural techniques. The actin-activated Mg+2 ATPase of rabbit skeletal muscle myosin can be inhibited by clam adductor paramyosin. Both proteins must be rapidly coprecipitated to form filaments for this inhibition. Slowly formed cofilaments are fully activatable by F-actin. In both cases, the cofilaments possess unique structural characteristics when compared to homofilaments.The mode of inhibition appears to be competitive when different concentrations of paramyosin and F-actin are compared. The apparent affinity of the myosin heads for actin is reduced by the presence of paramyosin within rapidly reconstituted thick filaments. These results suggest that paramyosin may serve as part of a relaxing mechanism within invertebrate muscles. It is unlikely that paramyosin plays a role in the initiation and maintenance of catch within specialized molluscan muscles.
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  • 94
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    Journal of Supramolecular Structure 3 (1975) 
    ISSN: 0091-7419
    Keywords: Life Sciences ; Molecular Cell Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
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  • 95
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    Journal of Supramolecular Structure 3 (1975), S. 181-191 
    ISSN: 0091-7419
    Keywords: Life Sciences ; Molecular Cell Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: The changes in birefringence in the rigor to relax transition of single Triton-extracted rabbit psoas muscle fibers have been investigated. The total birefringence of rigor muscle fibers was dependent on sarcomere length and ranged from (1.46 ± 0.08) × 10-3 to (1.60 ± 0.06) ± 10-3 at sarcomere lengths from 2.70 μm to 3.40 μm. An increase in total birefringence was measured dependent on sarcomere length when 55 single fibers were relaxed from the rigor state with Mg-ATP. Pyrophosphate relaxation produced a smaller increase in retardation when compared to Mg-ATP. The expected change in intrinsic birefringence during the rigor to relax transition was calculated assuming a hinge function of the subfragment 2 moiety of myosin. The changes in birefringence during isometric contraction and relaxation have been discussed in relation to possible structural changes.
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  • 96
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    Journal of Supramolecular Structure 3 (1975), S. 201-213 
    ISSN: 0091-7419
    Keywords: Life Sciences ; Molecular Cell Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Energy conservation and uncoupling in mitochondria are examined in the light of three important new findings: (a) Studies with the photoaffinity-labeling uncoupler 2-azido-4-nitrophenol have shown that mitochondria contain a specific uncoupler binding site (apparently a polypeptide of Mr = 30,000 ± 10%). (b) This site fractionates into an enzyme complex (complex V), which is capable of oligomycin- and uncoupler-sensitive ATP-Pi exchange. It is absent from electron transfer complexes I, III, and IV, which represent segments of the respiratory chain containing coupling sites 1, 2, and 3, respectively. (c) Trinitrophenol is a membrane-impermeable uncoupler (uncouples submitochondrial particles, but not mitochondria) and a poor protonophore. There is an excellent correlation between the uncoupling potencies and the affinities of uncouplers for the mitochondrial uncoupler-binding site. There is no correlation between uncoupling potency and protonophoric activity of uncouplers when a membrane-permeable uncoupler is compared with a membrane-impermeable one.
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  • 97
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    Journal of Supramolecular Structure 3 (1975), S. 192-199 
    ISSN: 0091-7419
    Keywords: Life Sciences ; Molecular Cell Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: The structural flexibility of creatine kinase has been investigated with the covalent hydrophobic probe 2-[4′-(2″-iodoacetamido) phenyl] aminonaphthalene-6-sulfonic acid (IAANS) which reacts at vastly different rates with the two subunits to give a protein conjugate with fluorescence characteristic of reaction with a site in a hydrophobic cleft. Binding of purine nucleotides greatly enhances the probe fluorescence while pyrimidine nucleotides quench the fluorescence. Small anions bind to nucleotide-free creatine kinase near the location of the transferable phosphoryl group and quench both the IAANS fluorescence of modified creatine kinase and the tryptophan fluorescence of native creatine kinase. Chloride and nitrate non-competitively inhibit MgADP binding both with and without creatine. Fluorescence energy transfer demonstrates that the active sites of creatine kinase are well separated and become further apart after the nucleotide-induced conformational change.
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  • 98
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    Journal of Supramolecular Structure 3 (1975), S. 214-221 
    ISSN: 0091-7419
    Keywords: Life Sciences ; Molecular Cell Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Evidence is presented that mitochondrial ATPase has two types of sites that bind adenine nucleotides. The catalytic site, C, binds the substrates ATP, GTP, or ITP and the inhibitor guanylyl imidodiphosphate (GMP-PNP). A second type of site, R, binds ATP, ADP, adenylyl imidodiphosphate (AMP-PNP), and the chromium complexes of ATP or ADP. All of these substances binding to the R site inhibit the hydrolysis of ATP in a competitive manner; their inhibition of hydrolysis of ITP and GTP is noncompetitive. GMP-PNP inhibits oxidative phosphorylation in submitochondrial particles but AMP-PNP does not. The localization on mitochondrial membranes of sites for the binding of various antibiotics that inhibit oxidative phosphorylation is discussed.
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  • 99
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    Journal of Supramolecular Structure 3 (1975), S. 231-241 
    ISSN: 0091-7419
    Keywords: Life Sciences ; Molecular Cell Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: The membrane bound coupling factor-latent ATPase was solubilized from the membrane vesicles of Mycobacterium phlei by using 0.25 M sucrose or low ionic strength buffer. Purification of the solubilized enzyme by use of Sepharose-ADP conjugate gel yielded a homogenous preparation of latent ATPase which was purified about 216-fold in a single step with an 84% yield. The enzyme exhibits a specific activity of 39 μmoles of ATP hydrolyzed per min per mg protein. The purified enzyme exhibits coupling factor activity. Electrophoresis in two dissociating solvent systems indicates that the enzyme contains at least three major polypeptides of molecular weights 56,000, 51,000 and 46,000 daltons, and two minor polypeptides of 30,000 and 17,000 daltons. Equilibrium binding studies of ADP with purified coupling factor-latent ATPase reveal the presence of two nucleotide binding sites per molecule with an apparent Ka of 8.1 × 10-5 M.By use of affinity chromatography, another latent ATPase has been isolated from the solubilized enzyme, which does not exhibit coupling factor activity.
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  • 100
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    Journal of Supramolecular Structure 3 (1975), S. 284-296 
    ISSN: 0091-7419
    Keywords: Life Sciences ; Molecular Cell Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: The tightly bound nucleotides of the beef-heart mitochondrial ATPase are released during cold inactivation followed by ammonium sulfate precipitation. During incubation at 0°C the sedimentation coefficient (s20 W) of the ATPase first declines from 12.1 S to 9 S. Prolonged incubation or precipitation with ammonium sulfate leads to dissociation of the 9 S component into subunits with s20 W of 3.5 S. The 9 S component still bears bound nucleotides which exchange more extensively and rapidly with added nucleotides than those bound to the active 12.1 S component. The bound nucleotides are lost when the 9 S form dissociates into the smaller subunits. Thus, firm binding of nucleotides is a property of the quarternary structure of the enzyme.The exchangeability of the nucleotides bound to the ATPase of chloroplast membranes is greatly increased in membranes illuminated in the presence of pyocyanine. Pi can exchange into both the β and γ positions of the bound nucleotides when the membranes are energized in the presence of Mg2+. The exchange of the nucleotides and the incorporation of Pi are insensitive to the inhibitor Dio-9 but are inhibited by the uncoupler S13.Abbreviation: S13, 5-chloro-3-t-butyl-2′-chloro-4′nitrosalicylanilide. This inhibition by S13 parallels that of the inhibition of photosynthetic phosphorylation. These findings are discussed with regard to our hypothesis that electron transfer causes release of preformed tightly bound ATP from the ATPase by inducing a conformational change.
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