Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts: Gene/Protein Disease Symptom Drug Enzyme Compound   Target Concepts: 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)

Formation of thick, stable foams and scums on activated sludge wastewater treatment plants is a worldwide problem, and to better understand what causes this foam and to cure it, there is a need to identify and quantify the bacteria present there. Fluorescence in situ hybridisation (FISH) overcomes the difficulties experienced with microscopic methods of identification for the mycolic-acid-containing actinomycetes (the mycolata), which are present in foams, where many share the morphotype of right-angled branching filaments. However, the presence of hydrophobic mycolic acids in their cell wall makes this group of bacteria particularly difficult to permeabilise, which greatly reduces the usefulness of FISH. While several permeabilisation treatments have been described, none appear to adequately permeabilise all genera of the mycolata. In this study several protocols for permeabilisation were assessed with both pure cultures of selected genera of the mycolata and foam samples. Combining mild acid hydrolysis with enzyme treatments (either mutanolysin/lysozyme or lipase/proteinase K) was found to be the most effective method, although other evidence presented here suggests that negative FISH results can not always be explained in terms of cell permeability to the probes.
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PMID:Improved permeabilization protocols for fluorescence in situ hybridization (FISH) of mycolic-acid-containing bacteria found in foams. 1567 95

Based on inorganic matrix controlled pore glass (CPG) and macro-pore silica sphere, by using polyethylene glycol (PEG 1000) as a ligand, a preparation method of hydrophobic interaction chromatographic (HIC) packing material was improved by adding a proper catalyst during the bonding process. The packing material can be synthesized in a scale-up batch, for example 150g for each batch, both for analytical and preparative columns. The retention of proteins, such as cytochrome C (Cyt-C), chymotrypsingen-A (Chy-A), lysozyme (Lys) and ribonuclease(Rnase), is increased with the increasing of (NH4)2SO4 concentration in the eluant 2.5 mol/L of salt concentration for the mobile phase was chosen by considering the separation efficiency and equipment life. After comparing the effect of pH for the retention of proteins it is found that the proteins are well separated at pH 7. The time of linear gradient elution program was optimized in considering the separation efficiency and speed. It is better to take 30 minutes of the gradient program for the separation. Six standard proteins can be well separated with the high-performance HIC column in the linear gradient elution program from 2.5 to 0 mol/L of (NH4)2SO4 in 50 mmol/L of phosphate buffer solution within 30 minutes. Cyt-C, Rnase, Lys and Chy-A can be separated by the HIC column based on CPG matrix. Six proteins, Cyt-C, Rnase, Lys, Chy-A, insulin(Ins) and lipase (Lip) can be well separated on the column based on silica matrix with gradient elution program. The recovery of trypsin detected with BAEE method is over 95% after purification with the HIC column.
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PMID:[Scale-up preparation of hydrophobic interaction chromatographic packing materials based on inorganic matrix]. 1573 63

Syringacin 4-A, a bacteriocin produced by Pseudomonas syrinagae 4-A, was obtained by induction with ultraviolet irradiation or mitomycin C. Approximately 1,000-fold purification of the bacteriocin was achieved by manganous chloride precipitation, differential centrifugation, and chromatography on hydroxyapatite columns. The purified syngacin was homogeneous on hydroxyapatite columns and sucrose density gradients; it also sedimented as a single entity in the analytical ultracentrifuge. The buoyant density of purified syringacin in cesium chloride was 1.294 g/ml. The sedimentation coefficient was calculated as 120S, and the diffusion coefficient was 6.49 x 10(-8) cm(2)/s. The molecular weight was calculated as 1.6 x 10(7) from physical data and 1.7 x 10(7) from biological data. The syringacin was composed of about 88.4% protein, 8.5% arabinose, 2.2% galacturonic acid, and 0.7% glucosamine. Amino acid analysis indicated a predominance of leucine (12.1%), aspartic acid (12.2%), and glutamic acid (12.7%). The ultraviolet spectrum showed a maximum absorbance peak at 276 nm. The syringacin was heat and alcohol sensitive, but resistant to trypsin, chymotrypsin, carboxypeptidase, Pronase, protease, lysozyme, steapsin, deoxyribonuclease, and ribonuclease. Maximum pH stability was between 5 and 8. Crude bacteriocin was stable at room temperature for at least a year, and purified material was stable for at least 3 months at 4 C.
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PMID:Purification and characterization of syringacin 4-A, a bacteriocin from pseudomonas syringae 4-A. 1582 74

The mouse-protective activity of Erysipelothrix rhusiopathiae culture supernatant fluids exists in a polydisperse form, ranging in density from aggregates which sediment at 10,000 x g for 3 hr to soluble units which will not sediment at 198,000 x g for 12 hr. A partially purified protective antigen has been isolated from the aggregates sedimented from a concentrate of the culture supernatant fluid at 20,000 x g for 3 hr. These aggregates contained the major protective antigen or antigens of E. rhusiopathiae, since, in addition to inducing active immunity, they adsorbed essentially all of the passively protecting antibody from rabbit antiserum produced by immunization with whole culture. The protective activity in these aggregates was destroyed by trypsin and greatly diminished by muramidase and heating at 64 C, but was not affected by lipase or ribonuclease.
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PMID:Isolation and Characterization of a Protective Antigen-Containing Particle from Culture Supernatant Fluids of Erysipelothrix rhusiopathiae. 1655 45

Three antigenic fractions from the cell walls of eight strains of mycobacteria were studied. Isolation and purification of these antigens were effected by enzymatic digestions, differential and sucrose gradient centrifugations, dialyses, and column chromatography. Two of the fractions were termed cell wall tuberculins (CWT-1, solubilized with lipase; CWT-2, solubilized with lysozyme); the third was termed "C" (cross-reacting) antigen. All appeared to be lipopolysaccharides. The CWT antigens, as compared with purified protein derivatives (human), were relatively species (group)-specific in both double immunodiffusion and guinea pig skin tests; in the latter, the reactions resembled those of delayed hypersensitivity. The C antigens reacted heterologously in double immunodiffusion and skin tests; the latter were the "immediate" type of reaction.
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PMID:Antigens of mycobacterial cell walls. 1655 48

The biological activity (D-value determination) of eggshell membrane (ESM) was examined to determine the membrane components and mechanisms responsible for antibacterial activity. Biological and enzymatic activities (i.e., beta-N-acetylglucosaminidase [beta-NAGase], lysozyme, and ovotransferrin) of ESM denatured with trypsin, lipases, or heat were compared with those of untreated ESM. Trypsin-treated ESM lost all biological activity (D-values at 54 degrees C were 5.12 and 5.38 min for immobilized and solubilized trypsin, respectively) but showed no significant loss of enzymatic activities. Treatments with porcine lipase and a lipase cocktail did not impact biological or enzymatic activities. Heat denaturation of ESM (at 80 and 100 degrees C for 15 min) resulted in significant decreases in biological activity (D-values of 3.99 and 4.43 min, respectively) and loss of beta-NAGase activity. Lysozyme and ovotransferrin activities remained but were significantly reduced. Purified ESM and hen egg white components (i.e., beta-NAGase, lysozyme, and ovotransferrin) were added to Salmonella Typhimurium suspensions (in 0.1% peptone water) at varying concentrations to evaluate their biological activity. D-values at 54 degrees C were 4.50 and 3.68 min for treatment with lysozyme or beta-NAGase alone, respectively, and 2.44 min for ovotransferrin but 1.47 min for a combination of all three components (similar to values for ESM). Exposure of Salmonella Typhimurium cells to a mixture of ovotransferrin, lysozyme, and beta-NAGase or ESM resulted in significant increases in extracellular concentrations of Ca2+, Mg2+, and K+. Transmission electron microscopic examination of Salmonella Typhimurium cells treated with a combination of ovotransferrin, lysozyme, and beta-NAGase revealed membrane disruption and cell lysis. The findings of this study demonstrate that ovotransferrin, lysozyme, and beta-NAGase are the primary components responsible for ESM antibacterial activity. The combination of these proteins and perhaps other ESM components interferes with interactions between bacterial lipopolysaccharides, sensitizing the outer bacterial membrane to the lethal affects of heat and possibly pressure and osmotic stressors.
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PMID:Identifying the components in eggshell membrane responsible for reducing the heat resistance of bacterial pathogens. 1662 12

We have studied the thermal stability of the triglyceride-hydrolyzing enzyme cutinase from F. solani pisi at pH values straddling the pI (pH 8.0). At the pI, increasing the protein concentration from 5 to 80 microM decreases the apparent melting temperature by 19 degrees C. This effect vanishes at pH values more than one unit away from pI. In contrast to additives such as detergents and osmolytes, the hydrophobic fluorophore 1,8-ANS completely and saturably suppresses this effect, restoring 70% of enzymatic activity upon cooling. ANS binds strongly to native cutinase as a noncompetitive inhibitor with up to 5 ANS per cutinase molecule. Only the first ANS molecule stabilizes cutinase; however, the last 4 ANS molecules decrease Tm by up to 7 degrees C. Similar pI-dependent aggregation and suppression by ANS is observed for T. lanuginosus lipase, but not for lysozyme or porcine alpha-amylase, suggesting that this behavior is most prevalent for proteins with affinity for hydrophobic substrates and consequent exposure of hydrophobic patches. Aggregation may be promoted by a fluctuating ensemble of native-like states associating via intermolecular beta-sheet rich structures unless blocked by ANS. Our data highlight the chaperone activity of small molecules with affinity for hydrophobic surfaces and their potential application as stabilizers at appropriate stoichiometries.
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PMID:pH-dependent aggregation of cutinase is efficiently suppressed by 1,8-ANS. 1696 99

The predicted amino acid sequence of Bacillus subtilis ycsK exhibits similarity to the GDSL family of lipolytic enzymes. Northern blot analysis showed that ycsK mRNA was first detected from 4 h after the onset of sporulation and that transcription of ycsK was dependent on SigK and GerE. The fluorescence of the YcsK-green fluorescent protein fusion protein produced in sporulating cells was detectable in the mother cell but not in the forespore compartment under fluorescence microscopy, and the fusion protein was localized around the developing spores dependent on CotE, SafA, and SpoVID. Inactivation of the ycsK gene by insertion of an erythromycin resistance gene did not affect vegetative growth or spore resistance to heat, lysozyme, or chloroform. The germination of ycsK spores in a mixture of L-asparagine, D-glucose, D-fructose, and potassium chloride and LB medium was also the same as that of wild-type spores, but the mutant spores were defective in L-alanine-stimulated germination. In addition, zymogram analysis demonstrated that the YcsK protein heterologously expressed in Escherichia coli showed lipolytic activity. We therefore propose that ycsK should be renamed lipC. This is the first study of a bacterial spore germination-related lipase.
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PMID:A novel lipolytic enzyme, YcsK (LipC), located in the spore coat of Bacillus subtilis, is involved in spore germination. 1722 Feb 30

The mass of air breathed by a human per day is equivalent to 10-times the mass of food consumed in that time. However, fundamental safety measures for atmospheric bacterial control have not yet been implemented. The purpose of our research is to develop a cell wall Iytic filter using a cell wall Iytic enzyme, which can inactivate the bacteria in air that cause infectious diseases by decomposing their cell envelope. In this study, the use of Iytic enzyme mixture was suggested, including glycosidase, protease and lipase. The performance of the Iytic enzyme mixture was evaluated using lysozyme, a typical Iytic enzyme, as a control. The substrate that we used was Micrococcus luteus, a gram-positive bacteria. The experimental results showed that the use of the Iytic enzyme mixture exhibited a Iytic rate per hour that was 13 - 39% greater than the control. Furthermore, although there are some different phases during bacterium multiplication, the Iytic rate per hour improved for all of the phases when the Iytic enzyme mixture was used.
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PMID:Inactivation of atmospheric bacteria using lytic enzyme mixture. 1727 36

Microbial carotenoids are difficult to extract because of their embedding into a compact matrix and prominent sensitivity to degradation. Especially for carotenoid analysis of bacteria and yeasts, there is lack of information about capability, precision and recovery of the method used. Accordingly, we investigated feasibility, throughput and validity of a new small-scale method using Micrococcus luteus and Rhodotorula glutinis for testing purposes. For disintegration and extraction, we combined primarily mild techniques: enzymatically we used combinations of lysozyme and lipase for bacteria as well as lyticase and lipase for yeasts. Additional mechanical treatment included sonication and freeze-thawing cycles. Chemical treatment with dimethylsulfoxide was applied for yeasts only. For extraction we used a methanol-chloroform mixture stabilized efficiently with butylated hydroxytoluene and alpha-tocopherol. Separation of compounds was achieved with HPLC, applying a binary methanol/tert-butyl methyl ether gradient on a polymer reversed C30 phase. Substances of interest were detected and identified applying a photodiode-array (PDA) and carotenoids quantitated as all-trans-beta-carotene equivalents. For evaluation of recovery and reproducibility of the extraction method, we used beta-8'-apo-carotenal as internal standard. The method provides a sensitive tool for the determination of carotenoids from bacteria and yeasts and also for small changes in carotenoid spectrum of a single species. Corequisite large experiments are facilitated by the high throughput of the method.
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PMID:A small-scale method for quantitation of carotenoids in bacteria and yeasts. 1750 7


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