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)

Protoplasts were prepared from Streptococcus sanguis and some S. mutans serotypes by use of lysozyme (EC 3.2.1.17) under particular conditions: cells had to be grown in DL-threonine (20 mM) and harvested in early exponential phase. The efficiency of protoplast formation was enhanced by two additional steps: plasmolysis (in 12% PEG), prior to addition of lysozyme, and a swirling phase, after the enzymic action. This procedure allowed us to obtain clean protoplasts, with only 0.5% contamination by bacterial cell walls. Up to 90% protoplast lysis was obtained in 0.5 M-NaCl. Cytoplasmic membrane purification was achieved by centrifugation on a glycerol cushion.
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PMID:Protoplast and cytoplasmic membrane preparations from Streptococcus sanguis and Streptococcus mutans. 636 Dec 17

An effective method for protoplast fusion in S. griseus producing grisin was developed. The method requires the use of polyethylene glycol with a molecular weight of 1000. It was demonstrated that protoplasts formed most effectively in this organism, when the mycelium of the streptomycete previously treated with ultrasound in the process of its growth was used for the treatment with lysozyme. The efficacy of protoplast regeneration in the strains with the use of the modified hypertonic medium R2MD was 25-75 per cent. The possibility of using the protoplast fusion method for constructing phage resistant strains producing kormogrisin was shown.
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PMID:[Use of the protoplast fusion method in the selection of Streptomyces griseus--the producer of the streptothricin antibiotic grisin]. 641 70

The high molecular weight penicillin-binding proteins (PBP(s) ) Bacillus subtilis PBPs 1, 2, and 4 and Bacillus stearothermophilus PBPs 1-4 were shown to catalyze peptidoglycan synthesis from the undecaprenol-containing lipid intermediate substrate in two assay systems. In a filter paper assay system, high levels of substrate polymerization occurred when reaction mixtures were incubated on Whatman 3MM filter paper. The pH optimum for peptidoglycan synthesis was 7.5 for B. subtilis PBPs 1, 2, and 4 and 8.5 for B. stearothermophilus PBPs 1-4. Polymerization was Mg2+-independent and was unaffected by sulfhydryl reagents. Reconstitution with membrane lipids or addition of detergent (optimal concentration, 0.1%) was necessary for synthesis to occur. Bacitracin, penicillin, and cephalothin did not affect polymerization while vancomycin, ristocetin, moenomycin, and macarbomycin were strong inhibitors. In a test tube assay system, optimal synthesis occurred either in the presence of 10% ethylene glycol, 10% glycerol, and 8% methanol or in the presence of 10% N-acetylglucosamine. The products of lysozyme digestion of the synthesized peptidoglycan were analyzed by gel filtration and paper chromatography. B. stearothermophilus PBPs 1-4 synthesized a peptidoglycan product that was 5-7% cross-linked. No evidence for cross-linking was apparent in the peptidoglycan product of B. subtilis PBPs 1, 2, and 4.
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PMID:Synthesis of peptidoglycan by high molecular weight penicillin-binding proteins of Bacillus subtilis and Bacillus stearothermophilus. 642 Apr 10

Nonmarine luminous bacteria belonging to the genus Vibrio cholerae were extremely sensitive to the bactericidal activity of human serum. Luminous bacteria incubated in a medium containing serum showed a decrease in their in vivo luminescence that was directly proportional to the decrease in the viable count and was a function of the serum concentration. Both immunoglobulins and the complement system were required to exert the serum bactericidal activity. Serum lacking immunoglobulins or certain complement components, especially C3, did not affect the luminescence. The bactericidal effect of the serum on luminous bacteria was diminished by the presence of lipopolysaccharide or by pretreatment of the serum with different species of killed bacteria. As found in other systems, the bacteriolytic activity of serum was only augmented by lysozyme, but was not lysozyme dependent; although the luminous bacteria were converted into spheroplasts in serum containing 0.5 M sucrose, their in vivo luminescence was almost not affected. This system could easily distinguish between the C classical pathway and the properdin pathway. Ethylene glycol-bis (beta-aminoethyl ether)-N,N'-tetraacetic acid, which inhibits only the classical complement pathway, did not inhibit the decrease in luminescence as did EDTA. Thus, it was possible to distinguish between deficiencies in complement components participating in both pathways and complement components that were involved only in the classical pathway. This system could also be used as a substitute to the hemolytic system in complement fixation tests.
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PMID:Determination of serum bactericidal activity with the aid of luminous bacteria. 661 81

Purified peptidoglycan (PG) obtained from Neisseria gonorrhoeae was tested for the ability to consume complement in normal human sera. Sonicated PG (S-PG), a heterogeneous mixture of soluble fragments (molecular weight, greater than 10(6)), as well as intact (insoluble) PG, reduced the level of whole hemolytic complement in a pool of four human sera. The minimal concentration of S-PG required for this activity was approximately 500 micrograms of S-PG per ml of serum. Complete lysozyme digestion of S-PG, yielding PG fragments of less than 10(4) molecular weight, eliminated complement-consuming activity. S-PG-mediated complement consumption resulted in depletion of the individual complement components C4 and C3. Consumption of complement did not occur when C4-deficient human serum or normal human sera treated with Mg2+-(ethylene glycol-bis(beta-aminoethyl ether)-N,N-tetraacetic acid to specifically impair classical complement pathway activity were used. The addition of rabbit anti-PG antibody greatly enhanced gonococcal PG-mediated complement consumption. Together, the data suggested that gonococcal PG-mediated complement consumption occurred via the classical complement pathway, was dependent on the presence of anti-PG antibody, and required glycosidically linked polymers of PG. Individual human sera varied widely in the extent of gonococcal PG-mediated reduction of complement levels, presumably a reflection of either different amounts of natural antibody to gonococcal PG, different levels of human PG hydrolase(s) capable of degrading PG to inactive fragments, or both.
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PMID:Complement consumption gonococcal peptidoglycan. 679 3

A method developed for the lysis of oral streptococci that employed the action of lysozyme suspended in dilute tris(hydroxymethyl)aminomethane-hydrochloride buffer containing polyethylene glycol has been adapted for use with lactobacilli, actinomycetes, propionibacteria, and pediococci. Most of the cellular deoxyribonucleic acid was liberated from many strains of bacteria usually thought to be lysozyme resistant. The major observations were as follows: (i) supplementation of the growth medium with L-threonine, L-lysine, or both frequently produced cells that were more susceptible to lysis by lysozyme; (ii) glucose-containing media produced cells that were more easily lysed than those from cultures grown on other substrates; (iii) polyethylene glycol not only served as an osmotic stabilizer, it also enhanced the extent of lysis; and (iv) dilute tris(hydroxymethyl)aminomethane buffer was superior to the buffer systems most commonly employed in published muramidase-based lysis techniques. Stationary-phase cells of Lactobacillus casei and Streptococcus mutans were more easily lysed than those isolated from log-phase cultures. The method as detailed in this report should be generally applicable for the lysis of gram-positive, asporogenous bacteria.
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PMID:Method for the lysis of Gram-positive, asporogenous bacteria with lysozyme. 698 47

The experimental conditions for plasmid transfer and genetic recombination in Staphylococcus aureus and some coagulase-negative staphylococci by protoplast fusion are described. Protoplasts were prepared by treatment with lysostaphin and lysozyme in a buffered medium with 0.7 to 0.8 M sucrose. Regeneration of cell walls was accomplished on a hypertonic agar medium containing succinate and bovine serum albumin. Transfer of plasmids occurred after treatment of the protoplast mixtures with polyethylene glycol (molecular weight, 6,000) not only between strains of the same species but also between parents of different species, although at approximately 100 times lower frequency in the latter case. Recombination of the chromosomal genes in fused protoplasts required simultaneous treatment of the mixed protoplasts with polyethylene glycol and CaCl2. A method was developed for isolation of recombinants after fusion between mutants of S. areus carrying unselectable markers. Antibiotic resistance plasmids were introduced into the parental strains and used as primary markers to detect protoplast fusion. Chromosomal recombinants were found among the clones with both parental plasmids at a high frequency. The method appears to have simple applications in the construction of strains with multiple mutant characters.
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PMID:Plasmid transfer and genetic recombination by protoplast fusion in staphylococci. 700 33

In order to clarify the mechanism of polyol-induced stabilization of protein, the thermal denaturation of lysozyme was studied at pH 4 in aqueous mixtures of some polyols (ethylene glycol, glycerol, erythritol, xylitol, and sorbitol) by a differential scanning calorimetry (DSC). The denaturation temperature, Td, increased with increasing the polyol concentration and the number of hydroxymethyl groups per polyol molecule. The calorimetric enthalpy or denaturation, delta H cal, increased with the increase in polyol concentration, but it was not significantly affected by the chain length of the polyol: delta H cal was about 30 kcal/mol larger in 30% (w/w) aqueous polyols than in water. The standard thermodynamic parameters for denaturation, delta G degrees, delta S degrees, and delta H degrees, which were calculated for glycerol and sorbitol systems using Td and delta H cal and assuming a constant heat capacity change, were an increasing function of polyol concentration. According to the thermodynamics of three component systems, it appeared that one or two polyol molecules are preferentially excluded from the domain of this protein on thermal denaturation. These thermodynamic data support the hypothesis that the thermal stabilization of lysozyme by polyols is due to a preferential solvent interaction effect which strengthens the hydrophobic interaction of the protein.
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PMID:Calorimetric study on thermal denaturation of lysozyme in polyol-water mixtures. 709 84

To facilitate the study of mechanisms and patterns of proteinuria, radioimmunoassays for human lysozyme (LZM) and albumin (alb) were established to permit quantitation of physiologic amounts of these proteins in urine. Commercially available LZM and alb preparations were radiolabeled with I125, and single antibody, competitive protein binding assays were developed. Separation of free and antibody-bound radioprotein was achieved with 20 per cent polyethylene glycol. LZM and alb 24-hr excretion rates for 12 normal subjects were 7 to 64 microgram and 2.3 to 16.1 mg, respectively. Of 6 renal disease patients with undetectable urine LZM by bioassay, 5 were shown to have elevated LZM concentrations by radioimmunoassay. The ease of establishing and performing these assays and their reproducibility suggest that they may have clinical and investigative value.
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PMID:Lysozyme and albumin radioimmunoassays. New techniques for the study of proteinuria. 737 42

A method has been developed to transform plasmid deoxyribonucleic acid into protoplasts of the insect pathogen Bacillus thuringiensis. Protoplasts were formed by treatment of cells with lysozyme. The efficiency of formation of protoplasts was affected by the strain, the media, and the cell density. Deoxyribonucleic acid uptake was induced by polyethylene glycol. Deoxyribonucleic acid from the Staphylococcus aureus plasmid pC194 was used for transformation. Although this plasmid could not be isolated as a stable extrachromosomal element, its chloramphenicol resistance was transferred to the recipient protoplasts. This was confirmed by assay for the enzyme chloramphenicol acetyltransferase, which confers resistance to chloramphenicol. This suggested that pC194 acts as an insertion element in B. thuringiensis.
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PMID:Transformation of Bacillus thuringiensis protoplasts by plasmid deoxyribonucleic acid. 746 65


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