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PubMed1983

Molecular properties of succinate dehydrogenase isolated from Micrococcus luteus (lysodeikticus).

Succinate dehydrogenase (EC 1.3.99.1) of Micrococcus luteus was selectively precipitated from Triton X-100-solubilized membranes by using specific antiserum. The precipitated enzyme contained equimolar amounts of four polypeptides with apparent molecular weights of 72,000, 30,000, 17,000, and 15,000. The 72,000 polypeptide possessed a covalently bound flavin prosthetic group and appeared to be strongly antigenic as judged by immunoprinting experiments. Low-temperature absorption spectroscopy revealed the presence of cytochrome b556 in the antigen complex. By analogy with succinate dehydrogenase purified from other sources, the 72,000 and 30,000 polypeptides were considered to represent subunits of the succinate dehydrogenase enzyme, whereas one (or both) of the low-molecular-weight polypeptides was attributed to the apoprotein of the b-type cytochrome. A succinate dehydrogenase antigen cross-reacting with the M. luteus enzyme complex could be demonstrated in membranes of Micrococcus roseus, Micrococcus flavus, and Sarcina lutea, but not in the membranes isolated from a wide variety of other gram-positive and gram-negative bacteria.

Cross Reactions↗
PubMed1986

Rapid lysostaphin test to differentiate Staphylococcus and Micrococcus species.

A rapid, simple lysostaphin lysis susceptibility test to differentiate the genera Staphylococcus and Micrococcus was evaluated. Of 181 strains from culture collections, 95 of 95 Staphylococcus strains were lysed, and 79 of 79 Micrococcus strains were not lysed. The seven Planococcus strains were resistant. Clinical isolates (890) were tested with lysostaphin and for the ability to produce acid from glycerol in the presence of erythromycin. Overall agreement between the methods was 99.2%. All clinical Micrococcus strains (43) were resistant to lysostaphin, and all clinical Staphylococcus strains (847) were susceptible. Seven of the Staphylococcus strains did not produce acid from glycerol in the presence of erythromycin. This lysostaphin test provides results in 2 h. It is easier to perform than previously described lysostaphin lysis methods. It is also more rapid and accurate than the glycerol-erythromycin test.

Bacteriological Techniques↗
PubMed1991

[Micrococcus luteus: a rare pathogen of valve prosthesis endocarditis].

A rare case of prosthetic valve endocarditis caused by Micrococcus luteus is described and compared with the few cases reported in the literature, as well as the clinical features, microbiological profile, therapy, and prognosis of common prosthetic valve endocarditis. Micrococcus luteus is a constituent of the normal human buccal bacterial flora which forms yellowish colonies and appears as a gram-positive coccus typically arranged in tetrades. Although of low virulence, the germ may become pathogenic in patients with impaired resistance, colonizing the surface of heart valves. In contrast to staphylococci (for which it may easily be mistaken) it is usually penicillin-sensitive. However, the most promising antibiotic regimen proposed for treatment of Micrococcus luteus seems to be a combination of vancomycin, amikacin, and rifampicin. If the infection leads to severe hemodynamic alterations, however, valve replacement may become necessary similar to the situation in prosthetic valve endocarditis caused by more aggressive and highly resistant bacteria.

Aged↗
PubMed1987

Accelerated tissue repair induced by micrococcus varians.

The present investigation was undertaken to study the effect of inoculated Micrococcus varians organisms on developing granulation tissue in rats. Subcutaneously implanted hollow cylindrical cellulose sponges were used as an inductive matrix for the growth of granulation tissue. The control implants were injected immediately after implantation with 1 milliliter of physiologic saline solution while the experimental implants were injected with a corresponding volume of saline solution containing live micrococci 10(7) microorganisms per milliliter. Cytologic and bacteriologic analyses of wound fluid aspirated from the central dead space of the implants were carried out three, seven and 14 days after implantation. Local blood flow and albumin extravasation were measured on day seven and granulation tissue grown into the implants was analyzed chemically on days seven and 14. No macroscopic infection with pus formation occurred, while Micrococcus varians was cultured from each inoculated implant. In the inoculated implants, the number of wound fluid neutrophils, granulation tissue blood flow and albumin extravasation increased significantly above the control level. Correspondingly, the amounts of granulation tissue deoxyribonucleic acid, nitrogen, collagen hydroxyproline, hexosamines and uronic acids in the inoculated implants exceeded significantly the control value on both days seven and 14. To conclude, inoculation of experimental wounds with nonpathogenic Micrococcus varians organisms enhanced local inflammatory reaction and blood flow, and promoted granulation tissue formation.

Animals↗
PubMedDate not supplied

[Use of an immunodiffusion analysis method in micrococcus species determination].

Eight species of the genus Micrococcus were studied in details for their antigenic specificities with the aid of the created bank of specific polyclonal antisera to the type strains of Micrococcus Cohn 1872--M. luteus CCM 169, M. varians CCM 884, M. roseus CCM 679 and M. nishinomiyaensis CCM 2140. Immunochemical analysis of 146 strains isolated from various natural and industrial substrata, as well as of 31 collection strains allowed us to reveal antigenic relatedness and distinctions between studied cultures and to accomplish taxonomic distribution of various Micrococcus species. By the immunoduffusion analysis the intraspecific antigenic relationship of M. roseus and M. varians was found, as well as significant antigenic heterogeneity of M. luteus. The results of antigenic analysis of micrococci may be used in the collection work and for the express-diagnostics of these microorganisms.

Antigens, Bacterial↗
PubMed1981

Reduced transcription activity of rat liver chromatin after protein restriction and selective digestion of nuclei with micrococcus nuclease.

Restriction of protein intake in the diet has been shown to produce a change in transcription activity as determined in vitro. Conditions for a reduced transcription activity were investigated with selective digestion of nuclei with micrococcus nuclease (EC 3.1.4.7). Liver nuclei from young male rats fed a diet containing either 20 or 3% casein for 6 days were digested with 1.3 microgram of micrococcus nuclease protein/mg of nuclear DNA (specific enzyme activity 15,000 or 150 units/mg of protein). Part of the chromatin-bound RNA polymerase activity was transferred from the 2,000 x g to the 102,000 x g pellet. Independent of the type of endonuclease use, the specific activity of chromatin-bound and soluble RNA polymerase I plus III was similar in the two groups of rats. After protein restriction RNA polymerase II activity was significantly diminished in the 2,000 x g pellet, and was unchanged in the 102,000 x g pellet. Heparin-stimulated and soluble RNA polymerase II activities were significantly reduced. Number and length of RNA chains synthetized by chromatin-bound RNA polymerase I plus III remained unchanged by dietary treatment. After a low protein diet, RNA polymerase II in the absence and presence of heparin synthesized an unchanged number of RNA chains with reduced length. A selective digestion of chromatin with micrococcus nuclease is needed to show the reduction in RNA polymerase II activity after protein restriction.

Animals↗
PubMed1983

Biotypes of Staphylococcus epidermidis and Micrococcus organisms, isolated from intramammary infections, reclassified into species of the genus Staphylococcus (epidermidis, hyicus, xylosus, and sciuri).

In a previous report (5), strains of Staphylococcus epidermidis and Micrococcus isolated from bovine intramammary infections were classified by the Baird-Parker (B-P) system and by serologic typing of the proteolytic enzymes. Since then, newer methods for distinguishing the genera Staphylococcus and Micrococcus and for defining new species have been reported. By utilizing these methods, the organisms from the original study were reclassified as follows: S. epidermidis B-P subgroup II, proteinase group F strains as S. epidermidis; S. epidermidis B-P subgroup III, proteinase group B strains as Staphylococcus hyicus subsp. chromogenes (pigmented) or subsp. hyicus (nonpigmented) and proteinase groups H and BH strains that were coagulase positive and nonpigmented as subsp. hyicus; B-P Micrococcus subgroup 6, proteinase group NR strains as Staphylococcus xylosus and subgroup 6, proteinase group G strains as Staphylococcus sciuri and S. xylosus. The reclassification of the strains increased the percentage of intramammary infections attributed to staphylococci from 62%, as first reported, to at least 86%.

Animals↗
PubMed1987

Pitted keratolysis. The role of Micrococcus sedentarius.

Though pitted keratolysis of the foot is generally viewed to be caused by bacteria, there is confusion regarding the identity of the causative organism. Species of Corynebacterium, Actinomyces, Dermatophilus, and Micrococcus have been proposed by various investigators. We have studied eight cases of pitted keratolysis and have cultured an organism identified as Micrococcus sedentarius on the basis of colonial morphology, micromorphology, biochemical reactions, and chemical analysis of whole-cell components. Furthermore, pitted keratolysis was produced experimentally in a human volunteer by applying M sedentarius under an occlusive dressing onto the surface of the heel.

Bacteriological Techniques↗
PubMed1995

Secretion of an antibacterial factor during resuscitation of dormant cells in Micrococcus luteus cultures held in an extended stationary phase.

A high proportion of Micrococcus luteus cells in cultures starved for 3-6 months in spent medium following growth to stationary phase in batch culture lost the ability to grow and form colonies on agar plates, but could be resuscitated from dormancy by incubation in liquid medium containing supernatant taken from the late log phase of viable cultures of the same organism (Kaprelyants et al. 1994). In the present work, we found that during the first 50-70 h of such resuscitation the dormant cells actually divide for 10-17 generations in lactate minimal medium containing yeast extract whilst remaining nonculturable on agar plates. Further incubation results in a decrease in the total cell number in liquid medium. The addition of viable (culturable) Micrococcus luteus cells in concentrations of up to 10(4) ml-1 to test tubes containing either resuscitating cells or supernatant from these cultures revealed the excretion of a factor or factors which inhibited the proliferation of otherwise viable cells. The maximum production of this factor took place after some 96 h of incubation of starved cells in resuscitation medium. Supernatant from late logarithmic phase batch cultures of M. luteus abolished the antibacterial effect of starved cultures incubated in resuscitation medium. It is concluded that the stimulating effect of viable cells, and of supernatant taken from batch cultures, on the resuscitation of dormant cells might be connected in part with overcoming the activity of an antibacterial factor causing self-poisoning of dormant cells during their resuscitation.

Anti-Bacterial Agents↗
PubMed1991

Identification of Staphylococcus and Micrococcus species with the STAPHYtest system.

A collection of 216 well-characterized strains of Staphylococcus, Micrococcus and Stomatococcus was examined by a commercially available STAPHYtest system (Lachema, Brno, Czechoslovakia). The results of STAPHYtest agreed with those of conventional tests. The STAPHYtest permitted a clear-cut separation of Staphylococcus from Micrococcus and Stomatococcus strains and correctly identified 104 of 145 (72%) Staphylococcus strains after 24 h of incubation. However, it allowed the identification only of 19 of 29 validly published Staphylococcus species. The STAPHYtest proved to be a simple and rapid system for the separation of staphylococci from micrococci and for the identification of most frequent clinically significant staphylococci.

Bacteriological Techniques↗
PubMed1975

Induction of nutritional mutants of Micrococcus glutamicus and their amino acid accumulation.

Micrococcus glutamicus ATCC 13032, a glutamic acid-producing organism, was treated with 0.2M ethylmethane sulfonate, the auxotrophs isolated showing varied patterns of extracellular amino acids. Eighty auxotrophic strains were obtained, out of which 31 excreted 1.0-4.0 mg threonine per ml and all the auxotrophs required biotin for growth and production of the amino acid. Eleven auxotrophs produced 1.5 to 3.0 mg alanine per ml and these auxotrophs required amino acids for their growth. Other auxotrophs lost their excretion capacity in subsequent fermentation trials. Further mutation of the biotin-requiring auxotroph Micrococcus glutamicus EM with gamma rays resulted in the isolation of 89 auxotrophic strains, out of which 28 excreted threonine (up to 5.0 mg per ml) higher than the parent auxotroph. Exposure to X-rays yielded 97 auxotrophs, out of these 35 producing 1.0-3.0 mg methionine per ml and requiring biotin for growth and production of the amino acid. Other auxotrophs produced alanine (0.5 to 2.0 mg per ml) and threonine (2.0 to 3.3 mg per ml). Irradiation with gamma rays favoured the development of threonine producing auxotrophs while X-rays favoured methionine-producing auxotrophs.

Alanine↗
PubMed1988

Elimination of mercury, cadmium and antibiotic resistance from Acinetobacter lwoffi and Micrococcus sp. at high temperature.

Resistance determinants for HgCl2 and CdCl2 were eliminated along with a number of antibiotic resistance factors from Acinetobacter lwoffi and Micrococcus sp. at 44 degrees C. These organisms were orginally resistant to HgCl2, merbromin, CdCl2, Pb(NO3)2, benzylpenicillin, erythromycin, carbenicillin, tetracycline and sulfadiazine. Four different types of mutants from A. lwoffi (type I to IV) and one type of mutant from Micrococcus sp. (type V) were obtained, depending on the loss of particular resistance factors for HgCl2, merbromin, CdCl2 and antibiotics. In general, frequency of elimination of all the missing markers was very low (in the range of 10(-3) per bacterium). However, the missing determinants did not revert spontaneously.

Acinetobacter↗
PubMed1975

Induction of mutation to streptomycin resistance in Micrococcus radiodurans.

No detectable induction of mutation to streptomycin resistance could be used in wild-type Micrococcus radiodurans and its radiation-sensitive and super-resistant mutants by ionizing or UV-radiation. N-methyl-N'-nito-N-nitrosoguanidine (NTG) was mutagenically active. The results suggest that repair of radiation-damaged DNA in Micrococcus radiodurans is mutation-proof.

Cobalt Radioisotopes↗
PubMed1994

Proteolytic and lipolytic activities of Micrococcus roseus (65), Halomonas elongata (16) and Vibrio sp. (168) isolated from Danish bacon curing brines.

Viable cells, cell free extracts and extracellular concentrates of Micrococcus roseus (65), Halomonas elongata (16) and Vibrio sp. (168) isolated from Danish bacon curing brines were examined for lipase, esterase, proteinase and aminopeptidase activities on natural and synthetic substrates. Micrococcus roseus (65) produced one intracellular esterase with affinity for short chain esters, and two intracellular aminopeptidases with affinity for nonpolar amino acids and L-arginine, respectively. One extracellular aminopeptidase with affinity for L-proline was also observed. Three intracellular esterases with affinity for short chain esters and a membrane bound esterase with affinity for butyric to capric esters were found in Halomonas elongata (16), but almost no aminopeptidase activity was found. Vibrio sp. (168) had four intracellular esterases with affinity for short chain esters and one esterase with affinity for all tested esters. Furthermore, an enzyme, which did not migrate on electrophoretic gels, had activity on all examined esters and tributyrin. Small intracellular activity on L-alanine was observed for this bacterial strain. There was no proteinase activities in the tested bacteria.

Aminopeptidases↗
PubMed1990

Micrococcus and Stomatococcus spp. from human infections.

Infections with Micrococcus spp. in six patients and Stomatococcus mucilaginosus in one patient are described. Two of the Micrococcus infections occurred in leukaemic patients with indwelling lines, six episodes occurred in three patients undergoing continuous ambulatory peritoneal dialysis and one occurred in a patient with a ventriculo-peritoneal shunt. Stomatococcus was isolated from fluid draining from a sub-dural haematoma. Colony morphology, oxidase reaction and resistance to nitrofuration were useful in differentiation of micrococci from staphylococci. Incidence and risk factors for micrococcal infections appear similar to those for infections with coagulase-negative staphylococci (CNS), but the literature is confused due to changes in the definitions of these genera.

Adolescent↗
PubMed1981

Successful immunotherapy with micrococcus, BCG or related polysaccharides on L1210 leukaemia after BCNU chemotherapy.

The experiments aimed at evaluating the optimal parameters in the chemo-immunotherapeutic treatment of the L1210 lymphoid leukaemia grafted to [female BALB/c (H2d) X male DBA/2 (H2d)]F1 hybrid mice, hereafter referred to as CDF1 mice. In vitro irradiation of leukaemic ascites cells by X- or gamma-rays and subsequent inoculation in mice showed that optimum immunogenicity is radiation dose-dependent. Grafting mice with 10(7) leukaemic ascites cells irradiated at optimum dose (80 GyX- or gamma-rays) delays mortality of the animals when challenged later with untreated L1210 cells, but is unable to cure mice. By contrast, specific immunoprophylaxis induced by Micrococcus, complement-triggering polysaccharides or BCG and irradiated leukaemic cells was able to protect mice against grafts of 10(4) L1210 cells. The i.p. route was notably superior to the i.v. route. When mice bearing advanced L1210 tumour were treated by chemotherapy (12 mg/kg of BCNU) on Day 6.5 after grafting 10(4) L1210 cells and subsequently treated by immunotherapy, a very high percentage (up to 90%) of mice with 10(8) leukaemic cells could be cured by repeated 1mg injections of bacterium or polysaccharide, and challenge with irradiated leukaemic cells was unnecessary. Because of the high cure rate obtained, the very regular response pattern and the non-pathogenicity, the bacterium Micrococcus lysodeikticus would seem a promising new candidate for chemo-immunotherapeutic antitumour strategies.

Animals↗
PubMed1974

Preparation and properties of a phosphoenzyme from the phosphoglucomutase of Micrococcus lysodeikticus.

1. Phosphoglucomutase from Micrococcus lysodeikticus was incubated with (14)C- and (32)P-labelled glucose 1,6-diphosphate and separated from the cofactor on a Sephadex column. (32)P-labelled phosphate (0.7mol/mol of enzyme) was associated with the enzyme, but no (14)C label was. 2. The (32)P-labelled enzyme exchanged its label with the substrates. When the labelled enzyme was incubated in Tris buffer, pH8.3, at 30 degrees C the proportion of exchangeable label slowly fell indicating a half-life of the phosphoenzyme of about 50h. 3. When HClO(4) was added to the labelled phosphoenzyme all of the label was precipitated with the protein and none was released as P(i). On alkaline hydrolysis P(i) was released at a rate comparable with the rate of hydrolysis of the phosphoenzyme from rabbit muscle. 4. We conclude that the phosphoenzyme from Micrococcus lysodeikticus yields a relatively stable, catalytically active phosphoenzyme when treated with cofactor, and that there is no evidence for the formation of an enzyme-glucose 1,6-diphosphate complex. The properties of the phosphoenzyme, which resemble those of rabbit muscle phosphoglucomutase, suggest that the phosphate may be bound to serine.

Animals↗
PubMed1980

Role of the subunits of the energy-transducing adenosine triphosphatase from Micrococcus lysodeikticus membranes studied by proteolytic digestion and immunological approaches.

An energy-transducing adenosine triphosphatase (ATPase, EC 3.6.1.3) that contains an extra polypeptide (delta) as well as three intrinsic subunits (alpha, beta, gamma) was purified from Micrococcus lysodeikticus membranes. The apparent subunit stoichiometry of this soluble ATPase complex is alpha 3 beta 3 gamma delta. The functional role of the subunits was studied by correlating subunit sensitivity to trypsin and effect of antibodies raised against holo-ATPase and its alpha, beta and gamma subunits with changes in ATPase activity and ATPase rebinding to membranes. A form of the ATPase with the subunit proportions 1.67(alpha):3.00(beta:0.17(gamma) was isolated after trypsin treatment of purified ATPase. This form has more than twice the specific activity of native enzyme. Other forms with less relative proportion of alpha subunits and absence of gamma subunit are not active. Of the antisera to subunits, only anti-(beta-subunit) serum shows a slight inhibitory effect on ATPase activity, but its combination with either anti-(alpha-subunit) or anti-(gamma-subunit) serum increases the effect. The results suggest that beta subunit is required for full ATPase activity, although a minor proportion of alpha and perhaps gamma subunit(s) is also required, probably to impart an active conformation to the protein. The additional polypeptide not hitherto described in Micrococcus lysodeikticus ATPase had a molecular weight of 20 000 and was found to be involved in ATPase binding to membranes. This 20 000-dalton component can be equated with the delta subunit of other energy-transducing ATPases and its association with the (alpha, beta, gamma) M. lysodeikticus ATPase complex appears to be dependent on bivalent cations. The present results do not preclude the possibility that the gamma subunit also plays a role in ATPase binding, in which, however, the major subunits do not seem to play a role.

Adenosine Triphosphatases↗