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The Journal of Biological Chemistry Oct 1995Although Micrococcus luteus UV endonuclease has been reported to be an 18-kDa enzyme with possible homology to the 16-kDa endonuclease V from bacteriophage T4 (Gordon,...
Although Micrococcus luteus UV endonuclease has been reported to be an 18-kDa enzyme with possible homology to the 16-kDa endonuclease V from bacteriophage T4 (Gordon, L. K., and Haseltine, W. A. (1980) J. Biol. Chem. 255, 12047-12050; Grafstrom, R. H., Park, L., and Grossman, L. (1982) J. Biol. Chem. 257, 13465-13474), this study describes three independent purification schemes in which M. luteus UV damage-specific or pyrimidine dimer-specific nicking activity was associated with two proteins of apparent molecular masses of 31 and 32 kDa. An 18-kDa contaminant copurified with the doublet through many of the chromatographic steps, but it was determined to be a homolog of Escherichia coli ribosomal protein L6. Edman degradation analyses of the active proteins yielded identical NH2-terminal amino acid sequences. The corresponding gene (pdg, pyrimidine dimer glycosylase) was cloned. The protein bears strong sequence similarities to the E. coli repair proteins endonuclease III and MutY. Nonetheless, traditionally purified M. luteus protein acted exclusively on cis-syn thymine dimers; it was unable to cleave site-specific oligonucleotide substrates containing a trans-syn -I, (6-4), or Dewar thymine dimer, a 5,6-dihydrouracil lesion, or an A:G or A:C mismatch. The UV endonuclease incised cis-syn dimer-containing DNA in a dose-dependent manner and exhibited linear kinetics within that dose range. Enzyme activity was inhibited by the presence of NaCN or NaBH4 with NaBH4 additionally being able to trap a covalent enzyme-substrate product. These last findings confirm that the catalytic mechanism of M. luteus UV endonuclease, like those of other glycosylase/AP lyases, involves an imino intermediate.
Topics: Amino Acid Sequence; Base Sequence; Cloning, Molecular; DNA Damage; DNA Glycosylases; DNA Primers; DNA, Bacterial; Deoxyribonuclease (Pyrimidine Dimer); Endodeoxyribonucleases; Escherichia coli Proteins; Genes, Bacterial; Imines; Micrococcus luteus; Molecular Sequence Data; Molecular Weight; Multienzyme Complexes; N-Glycosyl Hydrolases; Ribosomal Proteins; Sequence Homology, Amino Acid
PubMed: 7559510
DOI: 10.1074/jbc.270.40.23475 -
PloS One 2018The aim of the project was to find new catalysts capable of chlorolactone biotransformation. Three bicyclic chlorolactones with structures possessing one or two methyl...
The aim of the project was to find new catalysts capable of chlorolactone biotransformation. Three bicyclic chlorolactones with structures possessing one or two methyl groups in their cyclohexane ring were subjected to screening biotransformation using seven bacterial strains and one fungal strain from a salt mine. Three strains of bacteria (Micrococcus luteus Pb10, Micrococcus luteus WSP45, Gordonia alkanivorans Pd25) and one fungal strain (Aspergillus sydowii KGJ10) were able to catalyse hydrolytic dehalogenation of one substrate. The classification of the strains that were effective biocatalysts was confirmed by 16S rDNA analysis. The best result (76%) was obtained using Aspergillus sydowii KGJ10. All strains catalysed hydrolytic dehalogenation without changing the conformation. The equatorial position of the chlorine atom in the substrate turned out to be warrant of the positive result of the biotransformation process.
Topics: Aspergillus; Biocatalysis; Cyclohexanes; DNA, Ribosomal; Lactones; Micrococcus luteus; Mining; Sodium Chloride
PubMed: 29771957
DOI: 10.1371/journal.pone.0197384 -
BMC Research Notes Jun 2024The purpose of this study was to evaluate antibacterial activity of pigment extracted from bacteria, isolated from soil samples. During the study, 20 soil samples were...
The purpose of this study was to evaluate antibacterial activity of pigment extracted from bacteria, isolated from soil samples. During the study, 20 soil samples were collected from different areas (forest, agriculture fields, river sides and dumping sites) of Kathmandu and Lalitpur districts which were processed for isolation of pigment producing bacteria by spread plate technique. The pigmented bacterial isolates were identified and enriched in nutrient broth. Then, pigment was extracted in 95% methanol as solvent, which was further characterized using UV-Vis Spectrophotometric and TLC analysis. The obtained crude pigment extract was processed to carry out the antimicrobial susceptibility assay using agar well diffusion method. Out of 13 total pigmented bacteria isolates, four different colored pigmented bacterial isolates (S4O, S11Y, S14P and S17G) which produced efficient pigment on nutrient agar were chosen and they were further processed. Among these isolates, S4O was identified as Staphylococcus aureus, S11Y was identified as Micrococcus luteus, S14P was identified as Micrococcus roseus and S17G was identified as Pseudomonas aeruginosa respectively. On characterization using UV-Vis Spectrophotometric and TLC analysis, the pigment extracted from isolates S4O, S11Y and S14P were found to be Carotenoids and from isolate S17G was found to be Pyocyanin in nature. The maximum antibacterial activity was shown against Staphylococcus aureus from all the four pigments extracts. The green color pigment extract from isolate S17G was found to be most effective against all the Gram-positive and Gram-negative test bacteria. This study suggests that these pigment extracts from pigmented bacteria may have beneficial antibacterial roles that can be exploited in controlling unwanted bacterial growth.
Topics: Anti-Bacterial Agents; Soil Microbiology; Pigments, Biological; Microbial Sensitivity Tests; Staphylococcus aureus; Pseudomonas aeruginosa; Bacteria; Micrococcus luteus
PubMed: 38898523
DOI: 10.1186/s13104-024-06834-4 -
Journal of Bacteriology Dec 2019Bacteria have remarkable mechanisms to survive severe external stresses, and one of the most enigmatic is the nonreplicative persistent (NRP) state. Practically, NRP...
Bacteria have remarkable mechanisms to survive severe external stresses, and one of the most enigmatic is the nonreplicative persistent (NRP) state. Practically, NRP bacteria are difficult to treat, and so inhibiting the proteins underlying this survival state may render such bacteria more susceptible to external stresses, including antibiotics. Unfortunately, we know little about the proteins and mechanisms conferring survival through the NRP state. Here, we report that a universal stress protein (Usp) is a primary regulator of bacterial survival through the NRP state in NCTC 2665, a biosafety level 1 (BSL1) mycobacterial relative. Usps are widely conserved, and bacteria, including , , and , have multiple paralogs with overlapping functions that have obscured their functional roles. A kanamycin resistance cassette inserted into the universal stress protein A 616 gene (Δ::) ablates the UspA616 protein and drastically impairs survival under even short-term starvation (survival, 83% wild type versus 32% Δ::) and hypoxia (survival, 96% wild type versus 48% Δ::). We observed no detrimental UspA616 knockout phenotype in logarithmic growth. Proteomics demonstrated statistically significant log-phase upregulation of glyoxylate pathway enzymes isocitrate lyase and malate synthase in Δ:: We note that these enzymes and the UspA616 homolog (Rv2623) are important in virulence and chronic infection, suggesting that Usps are important stress proteins across diverse bacterial species. We propose that UspA616 is a metabolic switch that controls survival by regulating the glyoxylate shunt. Bacteria tolerate severe external stresses, including antibiotics, through a nonreplicative persistent (NRP) survival state, yet the proteins regulating this survival state are largely unknown. We show a specific universal stress protein (UspA616) controls the NRP state in Usps are widely conserved across bacteria, but their biological function(s) has remained elusive. UspA616 inactivation renders susceptible to stress: bacteria die instead of adapting through the NRP state. UspA616 regulates malate synthase and isocitrate lyase, glyoxylate pathway enzymes important for chronic infection. These data show that UspA616 regulates NRP stress survival in and suggest a function for homologous proteins in other bacteria. Importantly, inhibitors of UspA616 and homologs may render NRP bacteria more susceptible to stresses, including current antibiotics.
Topics: Bacterial Proteins; Citric Acid Cycle; Glyoxylates; Heat-Shock Proteins; Micrococcus luteus; Stress, Physiological
PubMed: 31548273
DOI: 10.1128/JB.00497-19 -
FEMS Immunology and Medical Microbiology Jan 1997Micrococcus luteus strains at a dose of 500 microg of whole cells caused anaphylactoid reactions leading to death in some instances within 1 h in C3H/HeN mice primed...
Micrococcus luteus strains at a dose of 500 microg of whole cells caused anaphylactoid reactions leading to death in some instances within 1 h in C3H/HeN mice primed with muramyl dipeptide (MDP, 100 microg). Tumor necrosis factor (TNF) and interleukin-6 (IL-6) were induced in the serum of half and of all the surviving mice, respectively. Cell wall specimens of M. luteus so far examined also caused anaphylactoid reactions accompanied by early death and one strain induced high levels of TNF and IL-6. Cytoplasmic membranes also induced IL-6. Essentially similar results were obtained with representative M. luteus cells and a cell wall specimen in MDP-primed C3H/HeJ mice. These results indicate that M. luteus has virulence activities that are associated with the induction of septic shock and systemic inflammatory diseases.
Topics: Acetylmuramyl-Alanyl-Isoglutamine; Anaphylaxis; Animals; Cell Wall; Cytokines; Female; Injections, Intravenous; Interleukin-6; Male; Mice; Mice, Inbred C3H; Micrococcus luteus; Tumor Necrosis Factor-alpha
PubMed: 9012443
DOI: 10.1111/j.1574-695X.1997.tb00995.x -
FEBS Open Bio Mar 2019The emergence of new antibiotic-resistant bacterial strains means it is increasingly important to find alternatives to traditional antibiotics, such as bacteriolytic...
The bacteriolytic activity of native and covalently immobilized lysozyme against Gram-positive and Gram-negative bacteria is differentially affected by charged amino acids and glycine.
The emergence of new antibiotic-resistant bacterial strains means it is increasingly important to find alternatives to traditional antibiotics, such as bacteriolytic enzymes. The bacteriolytic enzyme lysozyme is widely used in medicine as an antimicrobial agent, and covalent immobilization of lysozyme can expand its range of possible applications. However, information on the effect of such immobilized preparations on whole bacterial cells is quite limited. Here, we demonstrate the differential effects of glycine and charged (basic and acidic) amino acids on the enzymatic lysis of Gram-positive and Gram-negative bacteria by soluble and immobilized lysozyme. Glycine and basic amino acids (histidine, lysine, and arginine) significantly increase the rate of lysis of Gram-negative cells in the presence of soluble lysozyme, but they do not substantially affect the rate of enzymatic lysis of Gram-positive . Glutamate and aspartate significantly enhance enzymatic lysis of both and . When using immobilized lysozyme, the effects of amino acids on the rate of cell lysis are significantly reduced. For immobilized lysozyme, the presence of an external diffusion mode on cell lysis kinetics at bacterial concentrations below 4 × 10 colony-forming units·mL was shown. The broadening of the pH optimum of lysozyme activity after immobilization has been demonstrated for both Gram-positive and Gram-negative bacteria. The Michaelis constant () values of immobilized lysozyme were increased by 1.5-fold for cell lysis and 4.6-fold for cell lysis compared to soluble enzyme. A greater understanding of the effect of amino acids on the activity of native and immobilized lysozyme is important for both the development of new materials for medical purposes and elucidating the interaction of lysozyme with bacterial cells. Of particular interest is our finding that lysozyme activity against Gram-negative bacteria is enhanced in the presence of glycine and charged amino acids over a wide range of concentrations.
Topics: Amino Acids; Animals; Anti-Bacterial Agents; Chickens; Enzymes, Immobilized; Escherichia coli; Glycine; Microbial Sensitivity Tests; Micrococcus luteus; Muramidase; Particle Size; Sepharose; Surface Properties
PubMed: 30868059
DOI: 10.1002/2211-5463.12591 -
Microbiology Resource Announcements Jan 2019Micrococcus luteus has been found in a wide range of habitats. We report the complete genome sequence and methylome analysis of strain SA211 isolated from a hypersaline,...
Micrococcus luteus has been found in a wide range of habitats. We report the complete genome sequence and methylome analysis of strain SA211 isolated from a hypersaline, lithium-rich, high-altitude salt flat in Argentina with single-molecule real-time sequencing.
PubMed: 30701250
DOI: 10.1128/MRA.01557-18 -
Journal of Bacteriology Oct 1987The DNA sequence of the Micrococcus luteus str operon, which includes genes for ribosomal proteins S12 (str or rpsL) and S7 (rpsG) and elongation factors (EF) G (fus)... (Comparative Study)
Comparative Study
The DNA sequence of the Micrococcus luteus str operon, which includes genes for ribosomal proteins S12 (str or rpsL) and S7 (rpsG) and elongation factors (EF) G (fus) and Tu (tuf), has been determined and compared with the corresponding sequence of Escherichia coli to estimate the effect of high genomic G + C content (74%) of M. luteus on the codon usage pattern. The gene organization in this operon and the deduced amino acid sequence of each corresponding protein are well conserved between the two species. The mean G + C content of the M. luteus str operon is 67%, which is much higher than that of E. coli (51%). The codon usage pattern of M. luteus is very different from that of E. coli and extremely biased to the use of G and C in silent positions. About 95% (1,309 of 1,382) of codons have G or C at the third position. Codon GUG is used for initiation of S12, EF-G, and EF-Tu, and AUG is used only in S7, whereas GUG initiates only one of the EF-Tu's in E. coli. UGA is the predominant termination codon in M. luteus, in contrast to UAA in E. coli.
Topics: Amino Acid Sequence; Base Composition; Base Sequence; Cloning, Molecular; Codon; Cytosine; DNA, Bacterial; Escherichia coli Proteins; Genes, Bacterial; Guanine; Micrococcus; Molecular Sequence Data; Operon; Peptide Elongation Factor G; Peptide Elongation Factor Tu; Peptide Elongation Factors; Plasmids; Promoter Regions, Genetic; RNA, Messenger; Ribosomal Protein S9; Ribosomal Proteins; Sequence Homology, Nucleic Acid; Streptomycin; Transformation, Bacterial
PubMed: 3654584
DOI: 10.1128/jb.169.10.4770-4777.1987 -
Veterinary Research Feb 2021Tissue factor pathway inhibitors (TFPI), including TFPI-1 and TFPI-2, are Kunitz-type serine protease inhibitors that mainly inhibit the blood coagulation induced by...
Tissue factor pathway inhibitors (TFPI), including TFPI-1 and TFPI-2, are Kunitz-type serine protease inhibitors that mainly inhibit the blood coagulation induced by tissue factors. Previous reports on teleost proved TFPI play important roles in innate immunity. In this study, two TFPI (PoTFPI-1 and PoTFPI-2) molecules from Japanese flounder (Paralichthys olivaceus) were analyzed and characterized for their expression patterns, antibacterial and anticancer activities of the C-terminal derived peptides. Quantitative real time RT-PCR analysis shows that constitutive PoTFPI-1 expression occurred, in increasing order, in the brain, muscle, spleen, gills, head kidney, blood, intestine, heart, and liver; PoTFPI-2 was expressed, in increasing order, in the brain, gills, head kidney, muscle, intestine, spleen, liver, heart, and blood. Under the stimulation of fish pathogens, both PoTFPI-1 and PoTFPI-2 expressions increased significantly in a manner that depended on the pathogens, tissue type, and infection stage. Furthermore, C-terminal peptides TP25 and TP26, derived from PoTFPI-1 and PoTFPI-2, respectively, were synthesized and proved to be active against Micrococcus luteus (for TP25 and TP26) and Staphylococcus aureus (for TP25) via retardation effects on bacterial nucleic acids. In addition, TP25 and TP26 also displayed significant inhibitory effects on human colon cancer cell line HT-29. These results reveal that both PoTFPI-1 and PoTFPI-2 play important roles in host innate immunity. The antibacterial activity and anticancer cells function of TP25 and TP26 will add new insights into the roles of teleost TFPI.
Topics: Amino Acid Sequence; Animals; Anti-Bacterial Agents; Antineoplastic Agents; Fish Diseases; Fish Proteins; Flatfishes; Gene Expression Profiling; Gene Expression Regulation; Glycoproteins; Immunity, Innate; Lipoproteins; Micrococcus luteus; Phylogeny; Staphylococcus aureus
PubMed: 33632337
DOI: 10.1186/s13567-021-00908-y -
SpringerPlus 2014The screening of pollutant-degrading bacteria are limited due to most of bacteria in the natural environment cannot be cultivated. For the purpose of resuscitating and...
The screening of pollutant-degrading bacteria are limited due to most of bacteria in the natural environment cannot be cultivated. For the purpose of resuscitating and stimulating "viable but non-culturable" (VBNC) or uncultured bacteria, Micrococcus luteus proteins are more convenient and cost-effective than purified resuscitation-promoting factor (Rpf) protein. In this study, medium composition and culture conditions were optimized by using statistical experimental design and analysis to enhance protein production by M. luteus. The most important variables influencing protein production were determined using the Plackett-Burman design (PBD) and then central composite design (CCD) was adopted to optimize medium composition and culture conditions to achieve maximum protein yield. Results showed that the maximum protein yield of 25.13 mg/L (vs. 25.66 mg/L predicted) was obtained when the mineral solution, Lithium L-lactate, initial pH and incubation time were set at 1.5 ml/L, 8.75 g/L, 7.5 and 48 h, respectively. The predicated values calculated with the model were very close to the experimental values. Protein production was obviously increased with optimization fitting well with the observed fluorescence intensity. These results verified the feasibility and accuracy of this optimization strategy. This study provides promising information for exploring highly desirable pollutant-degrading microorganisms.
PubMed: 24616844
DOI: 10.1186/2193-1801-3-117