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)

Broadly cross-reactive monoclonal antibodies (MAbs) against enterobacterial outer membrane (OM) porin (Po) protein were isolated after immunization of BALB/c mice with whole cells of E. coli 055:B5. MAbs (n = 6) of the IgG class but of four different isotypes were studied. Based on a competition ELISA, all of the MAbs were directed against one and the same Po protein domain (Po I). The MAbs cross-reacted with 72 of 74 strains from 10 different genera of the Enterobacteriaceae. One Morganella and one Salmonella strain showed no cross-reactivity. Also, nine strains of various Neisseria spp. cross-reacted while 21 strains of various other nonenteric Gram-negative bacteria showed no cross-reactivity. The Po I sites were inaccessible in intact homologous bacteria but partially accessible in the OM. Digestion of OM with lysozyme or lysostaphin affected the accessibility of the Po I sites in OMs of various enterobacteria. Lysostaphin strongly enhanced the immunoaccessibility, whereas lysozyme had lesser effects. The enzymes also affected the binding by Neisseria OMs of the anti-Po I MAb. The Po I site was immunogenic both in humans and rabbits. The data indicate that Po I is an important Po protein domain, and that the effects of peptidoglycan-degrading enzymes must be considered in studies of Po protein domains.
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PMID:A conserved domain on enterobacterial porin protein analysed by monoclonal antibody. 184 63

In the present studies we compared the ability of two commonly used assays, solid-phase radioimmunoassay and enzyme-linked immunosorbent assay (ELISA), to detect human antibodies to Staphylococcus aureus peptidoglycan. ELISA was superior, with a reproducibility of 12.0%, as compared with 18.1% in solid-phase radioimmunoassay. Much lower serum dilutions could be used in ELISA. We also studied the effects of solubilizing the antigen by lysostaphin, lysozyme, or ultrasonication. Lysostaphin-treated peptidoglycan cannot be recommended since solid-phase radioimmunoassay could not distinguish positive from negative serum samples with this preparation. On the other hand, the sensitivity in both assays was high when peptidoglycan treated with lysozyme for 240 min or with ultrasonication for 30 min was used as antigen. The interassay correlation between solid-phase radioimmunoassay and ELISA was slightly better with sonicated peptidoglycan (correlation coefficient = 0.94, P less than 0.01), as compared with lysozyme-treated peptidoglycan (correlation coefficient = 0.76, P less than 0.01). We recommend the ELISA with sonicated peptidoglycan as antigen for use in routine serology.
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PMID:Methodological aspects of Staphylococcus aureus peptidoglycan serology: comparisons between solid-phase radioimmunoassay and enzyme-linked immunosorbent assay. 637 39

In this study we report the development of matrix-assisted laser desorption ionization mass spectrometry (MALDI-MS)-based methods for the structural characterization of muropeptides derived from peptidoglycan. Prior to analysis, peptidoglycan samples were subjected to enzymatic digestion with muramidase and the resulting muropeptides were purified by HPLC. A new matrix, 5-chloro-2-mercaptobenzothiazole, was employed for the MALDI-MS analysis. The results have demonstrated that sub-picomole to femtomole detection can be achieved in both positive mode and negative mode, allowing unambiguous determination of the molecular masses of monomeric and oligomeric muropeptides. Structural information from monomeric muropeptides was obtained by further postsource decay (PSD) analysis. Fragmentation patterns in positive mode and negative mode PSD were complementary for the elucidation of the peptide chain sequence. Lysostaphin digestion was also incorporated with MALDI mass mapping analysis for determination of peptide chain cross-linking patterns of muropeptide oligomers from Staphylococcus aureus strains.
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PMID:Structural characterization of peptidoglycan muropeptides by matrix-assisted laser desorption ionization mass spectrometry and postsource decay analysis. 917 19

The gene of microbial lysozyme (lyz) of S. aureus 118 and the gene of lysostaphin (lzf) of S. aureus RN 3239 were cloned and their expression in B. subtilis cells was shown. Lysozyme production in B. subtilis recombinant clone pLF14-Lyz, obtained as the result of cloning, was 2.5-fold greater than lysozyme production in S. aureus wild strain 118. Lysostaphin production in B. subtilis recombinant strain pLF14-Lzf which had inherited the cloned genes was approximately equal to lysostaphin production observed in S. aureus initial strain RN 3239. The production of lysozyme and lysostaphin in the cells of B. subtilis recombinant strains was observed at 30 degrees C and pH 5.5, while in S. aureus initial strains 118 and RN 3239 bacteria produced lysozyme and lysostaphin at 37 degrees C and pH 7.5 respectively.
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PMID:[Cloning of lysozyme and lysostaphin genes of Staphylococcus aureus and their expression in Bacillus subtilis cells]. 1156 57

Different steps and conditions for DNA extraction for microbiota analysis in sputum have been reported in the literature. We aimed at testing both dithiothreitol (DTT) and enzymatic treatments of sputum samples and identifying the most suitable DNA extraction technique for the microbiota analysis of sputum. Sputum treatments with and without DTT were compared in terms of their median levels and the coefficient of variation between replicates of both DNA extraction yield and real-time PCR for the 16S rRNA gene. Treatments with and without lysozyme and lysostaphin were compared in terms of their median levels of real-time PCR for S. aureus. Two enzyme-based and three beads-based techniques for DNA extraction were compared in terms of their DNA extraction yield, real-time PCR for the 16S rRNA gene and microbiota analysis. DTT treatment decreased the coefficient of variation between replicates of both DNA extraction yield and real-time PCR. Lysostaphin (either 0.18 or 0.36 mg/mL) and lysozyme treatments increased S. aureus detection. One enzyme-based kit offered the highest DNA yield and 16S rRNA gene real-time PCR with no significant differences in terms of alpha-diversity indexes. A condition using both DTT and lysostaphin/lysozyme treatments along with an enzymatic kit seems to be preferred for the microbiota analysis of sputum samples.
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PMID:How to Process Sputum Samples and Extract Bacterial DNA for Microbiota Analysis. 3034 4