Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: EC:3.2.1.17 (lysozyme)
 21,489 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

The cells of unicellular photosynthetic cyanobacterium Anacystis nidulans were permeated with lysozyme, toluene, toluene-triton, toluene-triton-lysozyme. Transmission electron microscopy of semi-thin sections (500 nm) using TEM at 160 kV showed that cells permeated with only lysozyme or toluene showed the typical concentric arrangement of thylakoid membranes. However, when toluene-treated cells were further treated with triton and lysozyme the thylakoid membranes were disrupted. Sequential reactions of Calvin cycle were studied in the differentially permeated cells in vivo, using various intermediates such as 3-PGA, GA-3-P, FDP, SDP, R-5-P, RuBP and cofactors like ATP, NADPH depending on the requirement. RuBP and R-5-P + ATP dependent activities could be observed in all types of permeated cells. Sequential reactions of the entire Calvin cycle using 3-PGA could be detected in the cells that had retained the internal organisation of the thylakoid membranes after permeation and were lost on disruption of this organisation. Light dependent CO2 fixation could be detected only in the cells permeated with lysozyme. This activity was abolished in the cells after treatment with toluene. The results suggested that the integrity of thylakoid membranes may be essential for the organisation of sequential enzymes of the Calvin cycle in vivo and facilitate their functioning.
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PMID:Involvement of thylakoid membranes in supramolecular organisation of Calvin cycle enzymes in Anacystis nidulans. 1268 42

Grau, F. H. (University of Wisconsin, Madison), and P. W. Wilson. Hydrogenase and nitrogenase in cell-free extracts of Bacillus polymyxa. J. Bacteriol. 85:446-450. 1963.-Washed cells of Bacillus polymyxa strain Hino, treated with lysozyme, yield cell-free extracts that rapidly evolve hydrogen from reduced methyl viologen, formate, and pyruvate. Hydrogenase is particulate, 86% being sedimented at 105,000 x g for 60 min. About 65% of the pyruvate metabolized is oxidized to acetyl phosphate, hydrogen, and carbon dioxide; the rest is converted to acetoin. These extracts fix considerable amounts of N(2) (15) when pyruvate is supplied as substrate, but will not fix with formate or mannitol. Centrifugation studies, and the absence of fixation with mannitol, show that this fixation is not caused by residual whole cells or spheroplasts. Cell-free fixation by B. polymyxa is similar to that by Clostridium pasteurianum. A short time lag in fixation occurs, and an optimal concentration of pyruvate is needed for maximal fixation. Arsenate causes a strong inhibition of fixation, presumably because arsenolysis of acetyl phosphate makes high-energy phosphate unavailable for the fixation process.
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PMID:Hydrogenase and nitrogenase in cell-free extracts of Bacillus polymyxa. 1394 23

The author has previously shown that the addition of egg-white lysozyme to the reaction mixture used in the Treponema pallidum immobilization test (TPI test) can reduce the time required for immobilization from 18 hours to as little as 6 hours. This opened up the possibility of developing a one-day TPI test and of further simplifying the procedure, as the conditions do not need to be so strictly controlled to ensure survival of the treponemes for the shorter time. In this paper it is shown that the standard procedure of incubation under an atmosphere of nitrogen and carbon dioxide can be replaced by incubation under a layer of liquid paraffin. The survival of treponemes is satisfactory under these conditions and the oil technique does not alter the sensitivity of the 6-hour test with added lysozyme. Comparative tests using the standard procedure, the lysozyme-gas technique and the lysozyme-oil technique showed that both modifications give the same degree of sensitivity and specificity after incubation for 6 hours as does the standard method. The feasibility of introducing these modifications into routine laboratory practice is discussed.
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PMID:A ONE-DAY TREPONEMA PALLIDUM IMMOBILIZATION TEST. 1431 9

In this study, various molecular dynamics simulations were conducted to investigate the effect of supercritical carbon dioxide on the structural integrity of hen egg white lysozyme. The analyses of backbone root-mean-square deviation, radius of gyration, and secondary structure stability all show that supercritical CO(2) exhibits the ability to increase the stability of this protein, probably as a result of the solvent with less polarity, where hydrophobic interactions stabilizing the native structure are weakened and simultaneously the local hydrogen bonds are strengthened, resulting in stabilization of the secondary structures. The hydrophobic cores in the alpha- and beta-domains also play an important role in preventing this protein from thermal unfolding. As supercritical CO(2) has been attractive for biomedical applications because of the advantages of mild critical condition, nonflammability, nontoxity, and the purity of the resulting products, the structural stabilizing effect found in this study strongly suggests that it is possible to increase the thermostability of hen egg white lysozyme by pretreatment with supercritical CO(2), leading to better industrial applications of this protein.
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PMID:Molecular dynamics simulations to determine the effect of supercritical carbon dioxide on the structural integrity of hen egg white lysozyme. 1517 1

A novel isoelectric precipitation of proteins in a pressurized carbon dioxide-water-ethanol system was developed where carbon dioxide was used as a volatile acid. The pH-pressure curves of the system with the absence and presence of proteins were investigated. By introducing the pressurized carbon dioxide to a solution containing protein, the pH value in the solution was decreased to the isoelectric region of the model protein BSA. Addition of ethanol could lower the buffer capacity of the protein, which made the precipitation concentration of protein go beyond the limits in a system without ethanol and well exploited the application field of the technique. In addition, ethanol in solution played the role of aiding precipitation in the process. Another model protein, hen egg white lysozyme, was also studied but could not be precipitated in the above system. All of these phenomena prove that isoelectric precipitation is the key point in the pressurized carbon dioxide-water-ethanol system.
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PMID:Novel isoelectric precipitation of proteins in a pressurized carbon dioxide-water-ethanol system. 1529 45

In this study, we use supercritical carbon dioxide as a processing medium for the fabrication of poly(DL-lactic acid) P(DLLA) microparticles that encapsulate a protein material. We have previously demonstrated that this polymer and a dry powder of a protein can be mixed under supercritical carbon dioxide conditions (above 31.1 degrees C and 73.8 bar) and that the protein component retains its biological activity. In this paper, we progress the work to demonstrate that the plasticized polymer and dry powder protein mixture can be sprayed to form solid polymer particles that encapsulate the protein. Particle size range is between 10 and 300 microm after spraying. Ribonuclease A and lysozyme were encapsulated in the polymer without significant loss of enzymatic activity. Biological assays of insulin and calcitonin confirm retention of activity after fabrication of the microparticles and release of the peptides/proteins.
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PMID:The production of protein-loaded microparticles by supercritical fluid enhanced mixing and spraying. 1558 96

Supercritical fluid (SCF) drying has been proposed as an alternative for freeze-drying to stabilize proteins. Here we studied the influence of sucrose and trehalose during SCF drying on the protein stability and the physical powder characteristics of lysozyme and myoglobin formulations. The results obtained with SCF drying were compared with the results after freeze-drying of the same solutions. Aqueous protein solutions, with or without sugar, were sprayed into a SCF mixture of carbon dioxide and ethanol. The dried products were analyzed by residual water measurements, scanning electron microscopy, X-ray powder diffraction and differential scanning calorimetry. After reconstitution the protein structure was studied by UV/VIS, circular dichroism and fluorescence spectroscopy, sodium dodecyl sulfate-polyacrylamide gel electrophoresis and bioactivity assay (lysozyme). The SCF dried and freeze-dried formulations showed comparable water contents, but their physical properties were substantially different. All freeze-dried cakes were amorphous with fully preserved protein structure. SCF dried sucrose-containing formulations showed agglomerated crystalline particles, whereas SCF dried trehalose-containing formulations appeared to consist of amorphous spherical particles. Particle morphology of excipients-free proteins was protein specific. Nearly all SCF dried lysozyme could be readily reconstituted, but for myoglobin significant fractions of SCF protein did not dissolve, especially in the absence of sugars. Covalent aggregation was not observed for the two proteins. For the recovered soluble fractions, the secondary protein structure was preserved. The tertiary structure was preserved for lysozyme, but not entirely for myoglobin. Surprisingly, during SCF drying trehalose was less protective than sucrose for myoglobin.
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PMID:Distinct effects of sucrose and trehalose on protein stability during supercritical fluid drying and freeze-drying. 1633 23

The surface activities of lysozyme and dipalmitoyl phosphatidylcholine (DPPC) vesicles at aqueous/compressed fluid interfaces are examined via high-pressure interfacial tension measurements using the pendant drop technique. The density and interfacial tension in compressible fluid systems vary significantly with pressure, providing a versatile medium for elucidating interactions between biomolecules and fluid interfaces and a method to elicit pressure-dependent interfacial morphological responses. The effects of lysozyme concentration (0.0008, 0.01, and 1 mg/mL) and pressure (> or = 7 MPa) on the dynamic surface response in the presence of ethane, propane, N2, and CO2 at 298 K were examined. Interfacial lysozyme adsorption reduced the induction phase and quickly led to interfacial tensions consistent with protein conformational changes and monolayer saturation at the compressed fluid interfaces. Protein adsorption, as indicated by surface pressure, correlated with calculated Hamaker constants for the compressed gases, denoting the importance of dispersion interactions. For DPPC at aqueous/compressed or aqueous/supercritical CO2 interfaces (1.8-20.7 MPa, 308 K), 2-3-fold reductions in interfacial tension were observed relative to the pure binary fluid system. The resulting surface pressures infer pressure-dependent morphological changes within the DPPC monolayer.
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PMID:Surface activity of lysozyme and dipalmitoyl phosphatidylcholine vesicles at compressed and supercritical fluid interfaces. 1637 53

A treatment procedure using lysozyme and ethylenediaminetetracetic acid gave intact but permeable cells (permeaplasts) of Anacystis nidulans. Rates of electron transport from water to carbon dioxide, ferricyanide, 2,6-dichlorophenol indophenol, benzoquinone, and methyl viologen, and from reduced indophenol to methyl viologen were measured as a function of treatment time. Rates of oxygen evolution in complete photosynthesis and electron flow from water to methyl viologen showed rapid and parallel decline with treatment time. Electron flow from water to ferricyanide and from reduced indophenol to methyl viologen increased during the first half hour of treatment (phase 1) to 60 to 80% of the original photosynthetic rate. Longer treatment (phase 2) resulted in decreased rate of ferricyanide reduction but not in rate of methyl viologen reduction from indophenol. Electron flow from water to quinone was two to three times higher than for complete photosynthesis in intact cells. It remained high during phase 1 and declined during phase 2. Phase 1 permeaplasts apparently retain high activity for photosystems 1 and 2 photoreactions.
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PMID:Photosynthetic properties of permaplasts of anacystis. 1665 14

In the perspective of production of dry therapeutic protein formulations, spray drying of lysozyme (as a model protein) into supercritical carbon dioxide was studied. The effects of the nozzle (i.e., co-current coaxial converging and converging-diverging, and T-mixer impinging) and process conditions (i.e., flow rates, pressure) on the drying of the lysozyme prepared in aqueous solution dried with supercritical carbon dioxide enriched with ethanol were investigated. The particle size distribution, width of particle size distribution and morphology were used to determine the effect of the various parameters assessed. Particles with a median size of approximately 1.5, approximately 5 or approximately 25 microns were produced depending of the nozzle selected. A basic comparative study of the nozzle was done by computational fluid dynamics, but the differences in particle size could not be depicted by these computations. The proportional increase of the flow rates (up to fivefold) caused a decrease in particle size (7- to 12-fold), and doubling the pressure caused a moderate decrease of the size (5-20%). The individual effect of the supercritical carbon dioxide, ethanol and solution streams was explained with a mass transfer model. Changing the ratio between flow rates slightly affected the particle size in various ways because of the swelling and shrinking stages of the drying droplet in supercritical carbon dioxide enriched with ethanol.
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PMID:Effect of the spraying conditions and nozzle design on the shape and size distribution of particles obtained with supercritical fluid drying. 1853 33


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