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Extracellular heme peroxidases in actinomycetes: a case of mistaken identity.

Actinomycetes secrete into their surroundings a suite of enzymes involved in the biodegradation of plant lignocellulose; these have been reported to include both hydrolytic and oxidative enzymes, including peroxidases. Reports of secreted peroxidases have been based upon observations of peroxidase-like activity associated with fractions that exhibit optical spectra reminiscent of heme peroxidases, such as the lignin peroxidases of wood-rotting fungi. Here we show that the appearance of the secreted pseudoperoxidase of the thermophilic actinomycete Thermomonospora fusca BD25 is also associated with the appearance of a heme-like spectrum. The species responsible for this spectrum is a metalloporphyrin; however, we show that this metalloporphyrin is not heme but zinc coproporphyrin. The same porphyrin was found in the growth medium of the actinomycete Streptomyces viridosporus T7A. We therefore propose that earlier reports of heme peroxidases secreted by actinomycetes were due to the incorrect assignment of optical spectra to heme groups rather than to non-iron-containing porphyrins and that lignin-degrading heme peroxidases are not secreted by actinomycetes. The porphyrin, an excretory product, is degraded during peroxidase assays. The low levels of secreted peroxidase activity are associated with a nonheme protein fraction previously shown to contain copper. We suggest that the role of the secreted copper-containing protein may be to bind and detoxify metals that can cause inhibition of heme biosynthesis and thus stimulate porphyrin excretion.

Actinomycetales↗

Distribution of thiols in microorganisms: mycothiol is a major thiol in most actinomycetes.

Mycothiol [2-(N-acetylcysteinyl)amido-2-deoxy-alpha-D-glucopyranosyl- (1-->1)-myo-inositol] (MSH) has recently been identified as a major thiol in a number of actinomycetes (S. Sakuda, Z.-Y. Zhou, and Y. Yamada, Biosci. Biotech. Biochem. 58:1347-1348, 1994; H. S. C. Spies and D. J. Steenkamp, Eur. J. Biochem. 224:203-213, 1994; and G. L. Newton, C. A. Bewley, T. J. Dwyer, R. Horn, Y. Aharonowitz, G. Cohen, J. Davies, D. J. Faulkner, and R. C. Fahey, Eur. J. Biochem. 230:821-825, 1995). Since this novel thiol is more resistant than glutathione to heavy-metal ion-catalyzed oxidation, it seems likely to be the antioxidant thiol used by aerobic gram-positive bacteria that do not produce glutathione (GSH). In the present study we sought to define the spectrum of organisms that produce MSH. GSH was absent in all actinomycetes and some of the other gram-positive bacteria studied. Surprisingly, the streptococci and enterococci contained GSH, and some strains appeared to synthesize it rather than import it from the growth medium. MSH was found at significant levels in most actinomycetes examined. Among the actinobacteria four Micrococcus species produced MSH, but MSH was not found in representatives of the Arthrobacter, Agromyces, or Actinomyces genera. Of the nocardioforms examined, Nocardia, Rhodococcus, and Mycobacteria spp. all produced MSH. In addition to the established production of MSH by streptomycetes, we found that Micromonospora, Actinomadura, and Nocardiopsis spp. also synthesized MSH. Mycothiol production was not detected in Propionibacterium acnes or in representative species of the Listeria, Staphylococcus, Streptococcus, Enterococcus, Bacillus, and Clostridium genera. Examination of representatives of the cyanobacteria, purple bacteria, and spirochetes also gave negative results, as did tests of rat liver, bonito, Candida albicans, Neurospora crassa, and spinach leaves. The results, which indicate that MSH production is restricted to the actinomycetes, could have significant implications for the detection and treatment of infections with actinomycetes, especially those caused by mycobacteria.

Actinomycetales↗

COMPARISON OF THE CELL-WALL COMPOSITION OF MORPHOLOGICALLY DISTINCT ACTINOMYCETES.

Yamaguchi, Tatsuro (The University of Tokyo, Tokyo, Japan). Comparison of the cell-wall composition of morphologically distinct actinomycetes. J. Bacteriol. 89:444-453. 1965.-Cell-wall composition of various morphologically distinct actinomycetes was studied to determine the relationship, if any, between cell-wall composition and morphological criteria in actinomycete taxonomy. The methods used were similar to those of Cummins and Harris. At least five types of cell-wall composition were obtained; however, these were not always correlated with groupings by the conventional classification system. For instance, the sporangium-forming actinomycetes, Actinoplanaceae, had three types of cell-wall composition; the composition of cell walls of Promicromonospora, Micromonospora, and Microbispora was the same as, or similar to, that of Actinomyces, Actinoplanes, and Streptosporangium, respectively; Chainia, Actinopycnidium, Actinosporangium, and Microellobosporia had the same cell-wall composition as Streptomyces, whereas that of Streptoverticillium was slightly different. Possible implications of cell-wall composition and morphological differentiation of hyphae for the taxonomy and phylogeny of actinomycetes are also discussed.

Actinobacteria↗

Characterization of exposure to molds and actinomycetes in agricultural dusts by scanning electron microscopy, fluorescence microscopy and the culture method.

Air samples from 79 farms with 10(5) to 10(11) microorganisms/m3 were analyzed by scanning electron microscopy (SEM), fluorescence microscopy (FM), and the culture method. The total exposure to microorganisms (particularly actinomycetes) was underestimated when assessed as colony-forming units (cfu). The average cfu count was one-sixth of the total count according to SEM or FM, and the individual variability was great. This occurrence was partly explained by the aggregation of spores. Single spores accounted for 2-65% of all spores in 35 samples. There was an average of three spores/particle, and 93 (range 67-100)% of the spores were single or in aggregates of respirable size. Aggregation was more pronounced for actinomycetes and at high spore counts. Actinomycetes and bacteria could not be distinguished by FM. Bacteria (other than actinomycetes) were not detected by SEM, yet the total count of microorganisms was similar for FM and SEM. Most particles were spores from actinomycetes and fungi of the genera Aspergillus or Penicillium.

Actinomycetales↗

[The application of succession analysis in combination with extremely high-frequency irradiation to the selective isolation of actinomycetes from soil].

A new method employing succession analysis and extremely high-frequency (EHF) irradiation is proposed for the selective isolation of actinomycetes from soil. Total actinomycetes were efficiently isolated from soil suspensions irradiated in the wavelength band 4.6-5.8 mm on the 14th and 45th days of succession initiated by soil wetting and from soil suspensions irradiated in the wavelength band 8-11.5 mm on the 7th day of succession. The rare actinomycete genera Actinomadura, Micromonospora, Nonomuraea, Microbispora, Amycolatopsis, Pseudonocardia, Saccharothrix, Streptosporangium, Actinosynnema, Nocardioides, and Saccharopolyspora were isolated by either of the two approaches (succession analysis and EHF irradiation); however, the range of isolated rare actinomycetes was considerably wider when the combination of the two approaches was used. For instance, actinomycetes of the rare genera Actinocorallia, Promicromonospora, Actinoplanes, and Kibdelosporangium were isolated only when EHF irradiation was employed at the early stages of succession.

Actinobacteria↗

[Number and structure of actinomycetes complexes in the rhizosphere winter rye, oat and red clover].

The actinomycetes complexes in the rhizosphere of three agricultural plants by using the methods of luminescense microscopy and cup sowing were investigated. It was established, that concentration of prokaryotic biomass and biomass of actinomycetes mycelium in rhizosphere of plants is higher than in free from the radicals to soil. Rhizosphera of the oat (Avena sativa L.) and red clover (Trifolium pratense L.) is colonized by Streptomyces, Micromonospora and olygospore species. Dominante actinomycetes of winter rye (Secale cereale L.) are classified into the genera Micromonospora. It was shown that numbers and biomass of actinomycetes mycelium were fond to decreased, diversity of actinomycetes in contrast is increased in the series: "winter rye--oat--red clover". In connection with ecological safety the capability of increase with prokaryotes naturally disease suppressive soil and stability of plants to pathogen is discussed.

Actinobacteria↗

[Numerical taxonomy of verticillate actinomycetes].

The paper discusses several methods used in numeric classification of verticillate actinomycetes. A method is suggested to estimate different significance of their characteristics. The use of the principle of nonequivalent characteristics has been found to increase the differentiating role of the most genetically stable traits, which makes it possible to reduce the number of necessary calculations using computers in the numeric taxonomy of actinomycetes. The production of polyene substances is the most informative property in the sampling of actinomycetes under study. The use of the methods of numeric taxonomy yields a system of actinomycetes that is most close to only one among the known systems for phenotypical classification of verticillate actinomycetes; therefore, the latter is a genuine one.

Computers↗

Cellulolytic activity of aerobic soil actinomycetes.

The cellulolytic activity of several aerobic soil actinomycetes against insoluble cellulose and soluble cellulose derivatives (CMC-carboxymethylcellulose) was studied. From the soil, 8 actinomycete strains were isolated after enrichment growth and purification on the same synthetic medium. The actinomycete strains were able to degrade insoluble cellulose, with the production of cellobiose and various oligosaccharide intermediates as degradation products, indicating the random attack of the cellulose chain. The actinomycete strains showed also a great activity against soluble cellulose (CMC). The viscosity of CMC solutions decreased rapidly and was followed by an increase in reducing compounds. The degree of substitution of CMC solutions had an effect on the degradation by the actinomycetes. The degree of polymerization did not affect the rate of hydrolysis, however.

Actinomycetales↗

Thermophilic actinomycetes associated with agro-environment of Punjab state (India).

Thermophilic actinomycetes (TAs) are unique high temperature aerobic bacteria which belong to the group actinomycetes. While working on hypersensitivity pneumonitis (HP), an immunological disorder, resulting from the inhalation of spores of thermophilic actinomycetes, it was considered worthwhile to record their occurrence in the environment of Punjab. Thermophilic actinomycetes isolated by spread plate method from nine different substrates of the Punjab environment were identified as 8 different species. Atmospheric occurrence of TAs was detected by exposing Petri dishes containing soy agar medium which were exposed in six different environments of the state. Seven thermophilic species were recorded. Thermoactinomyces vulgaris was the predominant species with 87.5, 87.1, and 100% prevalence in the soil, natural manure and the wheat field, respectively. The study is the first report of its kind from this region.

Actinomycetaceae↗

Distribution of clinically important thermophilic actinomycetes in vegetable substrates and soil in north-western India.

Medically important thermophilic actinomycetes were isolated from 218 (64%) of 341 samples of vegetable substrates and soil examined from sites in north-western India. Thermoactinomyces vulgaris (T. candidus) was the commonest species, occurring in 56% of samples, followed by Saccharomonospora viridis in 29%, Thermoactinomyces thalpophilus in 27%, Faenia rectivirgula (Micropolyspora faeni) in 21% and Thermoactinomyces sacchari in 14%. T. vulgaris and T. thalpophilus were isolated from all types of substrate examined, with T. vulgaris always more common than T. thalpophilus. Of the other thermophilic actinomycetes, F. rectivirgula was isolated predominantly from hay (44%) and S. viridis (56%) and T. sacchari (44%) from sugar-cane bagasse. The largest populations of T. vulgaris and T. thalpophilus were found in paddy straw, followed by T. sacchari, S. viridis and F. rectivirgula in sugar-cane bagasse. The widespread occurrence of these clinically important thermophilic actinomycetes suggests that exposure of humans and animals to them may be frequent in north-western India. Studies are required to determine the prevalence of extrinsic allergic alveolitis (hypersensitivity pneumonitis) caused by thermophilic actinomycetes in the local population.

Actinomyces↗

Thermophilic actinomycetes in cane sugar mills: an aeromicrobiologic and seroepidemiologic study.

Aerial prevalence of clinically important thermophilic actinomycetes and occurrence of precipitating antibodies against them in sera of 153 exposed workers have been reported. The study was carried out in two cane sugar mills namely, the Upper Doab Sugar Mills and the Ramala Sugar Mills, located in north-west India. In both the sugar mills, T. sacchari was the predominant species, it accounted for 55.1% and 50.3% of the total population of thermophilic actinomycetes, followed by T. vulgaris (19.7% and 23.7%), T. thalpophilus (21.1% and 17.1%), Saccharomonospora viridis (3.4% and 5.0%) and Saccharopolyspora rectivirgula (Faenia rectivirgula) (0.7% and 3.9%), respectively. Precipitating antibodies against thermophilic actinomycetes were demonstrable in 34 (22.2%) workers; T. sacchari alone accounted for 20 of the positive precipitin reactions, followed by S. rectivirgula in 10. The mean absorbance values for IgG antibody activity against T. sacchari as well as S. rectivirgula were found to be elevated significantly in the symptomatic workers than in the asymptomatic workers (p < 0.05) or unexposed controls (p < 0.001). However, the difference in IgG antibody activity was insignificant between precipitin-positive symptomatic workers and precipitin-positive asymptomatic workers. The results indicate that clinically important thermophilic actinomycetes are widely prevalent in cane sugar mills, and T. sacchari and S. rectivirgula are the major species involved in the sensitization of the bagasse workers in India.

Actinomycetales↗

Biosystematics and diversity amongst novel carboxydotrophic actinomycetes.

Fifty-four carboxydotrophic actinomycetes isolated from soils and composts were compared through 119 unit characters with representative mesophilic and thermophilic streptomycetes. The data were examined using the Jaccard, pattern and simple matching coefficients and clustering achieved using the unweighted pair group method with arithmetic averages algorithm. Acceptable cophenetic correlation and test error values allowed confidence to be placed in the resultant numerical taxonomies. The carboxydotrophic actinomycetes, which were distinct from cluster-groups corresponding to the mesophilic and thermophilic streptomycetes, formed two major cluster-groups the members of which were examined for the presence of diagnostic chemical markers. All but two of the carboxydotrophic actinomycetes had a profile of chemical properties consistent with their assignment to the genus Streptomyces. Quantitative fatty acid data were examined using the SIMCA package and the two statistically significant groups obtained corresponded with the cluster-groups circumscribed in the numerical phenetic analysis. Members of the two groups were also distinguished on the basis of their phospholipid composition. The two strains that contained meso-as opposed to LL-diaminopimelic acid in their peptidoglycan also showed a distinct chemotaxonomic profile. It was concluded that the carboxydotrophic actinomycetes form a novel and taxonomically diverse group.

Actinomycetales↗

New PCR primers for the screening of NRPS and PKS-I systems in actinomycetes: detection and distribution of these biosynthetic gene sequences in major taxonomic groups.

Nonribosomal peptide synthetases (NRPS) and type I polyketide synthases (PKS-I) are biosynthetic systems involved in the synthesis of a large number of important biologically active compounds produced by microorganisms, among others by actinomycetes. In order to assess the occurrence of these biosynthetic systems in this metabolically active bacterial group, we designed new PCR primers targeted to specifically amplify NRPS and PKS-I gene sequences from actinomycetes. The sequence analysis of amplified products cloned from two model systems and used to validate these molecular tools has shown the extreme richness of NRPS or PKS-I-like sequences in the actinomycete genome. When these PCR primers were tested on a large collection of 210 reference strains encompassing all major families and genera in actinomycetes, we observed that the wide distribution of these genes in the well-known productive Streptomyces species is also extended to other minor lineages where in some cases very few bioactive compounds have been identified to date.

Actinobacteria↗

Enzyme inhibitors and other bioactive compounds from marine actinomycetes.

Several enzyme-inhibitor-producing actinomycetes were isolated from various samples collected from the marine environment and characterized. Most of them produced novel compounds that are useful in medicine and agriculture. Actinomycete strain no. 18, which produces antibiotics against Gram-positive bacteria only in the presence of seawater, was isolated from sediment sampled from neritic sea water and characterized. The production of antibiotics was observed at seawater concentrations ranging from 60 to 110% (v/v). Thus, the production was seawater-dependent. The production of tetrodotoxin (TTX), known otherwise as puffer fish toxin, was investigated in various actinomycetes collected from the marine environment. Of 10 isolates from various sea areas, 9 produced TTX as judged by their retention times on high-performance liquid chromatography (HPLC). To our knowledge, this is the first report of actinomycetes from the marine environment that produce TTX.

Acetylglucosaminidase↗

Actinomycetes as agents of biodegradation in the environment--a review.

The diversity of form in the Actinomycetales is well-recognised, due to the sustained generation of environmental isolates for pharmaceutical screening. Actinomycetes isolated from soil and related substrates show primary biodegradative activity, secreting a range of extracellular enzymes and exhibiting the capacity to metabolise recalcitrant molecules. Composting is one process which relies heavily on such prolific actinomycete activity. Amongst actinomycetes in soil, there are examples of different strategies, from cycles of rapid proliferation and sporulation to the maintenance of populations by prolonged slow growth and scavenging, and the evidence for this is examined. The mechanisms of lignocellulose degradation by actinomycetes are discussed in relation to functional conservation within the group, and correlations with those described in other bacteria and fungi.

Actinomycetales↗

Actinomycete infections in humans--a review.

Diseases caused by pathogenic aerobic and facultatively anaerobic actinomycetes differ considerably with respect to their etiology, pathogenesis, clinical appearance and epidemiology. Facultatively anaerobic (fermentative) actinomycetes may not only be involved etiologically in the three classical forms of cervicofacial, thoracic and abdominal actinomycoses, but also in infections of the female genital organs, the eye, the tissue adjacent to dental implantation elements and tooth extraction wounds. The species distribution of the fermentative actinomycetes isolated from these conditions varied to a certain, but characteristic, extent, as did the concomitant actinomycotic flora. The sex ratio reported for human Actinomyces infections (male:female = 3:1) appeared to be restricted to actinomycotic abscesses and empyemas. The prevailing pathogenic, obligately aerobic actinomycete species in Germany was found to be Nocardia farcinica followed by Nocardia asteroides. The comparatively high incidence of N. farcinica infections was chiefly due to the occurrence of nosocomial postoperative wound infections by this pathogen observed in two German hospitals. Besides surgical treatment, immunosuppressive treatment appeared to be the most common factor predisposing for nocardiosis. Recent observations strongly suggested that the spectrum of human nocardial infections in Germany has been changing, as regards the overall incidence, the prevalence of N. farcinica, the sex ratio, the mean age of patients, as well as the role of N. farcinica as a possibly important nosocomial pathogen.

Actinomycetales Infections↗

Detection and identification of novel actinomycetes.

The actinomycetes are well known as a group of filamentous, Gram-positive bacteria that produce many useful secondary metabolites, including antibiotics and enzymes. Although they have been intensively studied for both theoretical and practical objectives, there is much scope for developing our basic knowledge of the means of detection and isolation of these microbes. This session concentrated on new methods for the detection and identification of novel actinomycetes from a range of environments. Approaches to the detection of actinomycetes ranged from investigations of neglected habitats and extreme environments (e.g. alkaline soils and oil drills) to the analysis of DNA extracted from the environment and use of specific phages. The continuing problems of the identification of actinomycete isolates were also considered. Topics discussed included use of phage typing, DNA probes, and correlation between phenetic and genotypic species of Streptomyces.

Actinomycetales↗

Novel bioactive compounds from actinomycetes.

Actinomycetes form an enormous reservoir of secondary metabolites and enzymes. The potential for exploiting rare actinomycetes is highlighted by the discovery of novel compounds from strains of Spirillospora and Nocardioides. Novel compounds of well known classes of antibiotics, such as polyenes, continue to be discovered. For compounds containing a chromophore, the analysis by high-performance liquid chromatography coupled with a diode-array detector enables the elimination of producers of known compounds and facilitates the discovery of novel compounds or derivatives. The complexity of the regulatory mechanisms is illustrated by glutamine synthetase. The characterization of thermostable amylolytic, lignolytic, peroxidase and neuramidase activities, and the isolation of novel cellulolytic actinomycetes clearly demonstrate the potential of Actinomycetes as producers of enzymes.

Actinomycetales↗