The protective role of pigments against UV rays in fungi isolated from the mesosphere

Five among six species of microorganisms isolated from the mesosphere contained pigments which made them more resistant to the action of UV as compared to pigmentless microorganisms in the atmosphere of Earth. UV irradiation in the atmosphere is supposed to select resistant pigmented forms, so that they predominate in the mesosphere. To confirm this assumption, mutants of Aspergillus niger, Penicillium notatum and Circinella muscae were sported by irradiating them four times and then subjecting to stepwise selection. These mutants either synthesized pigments at a very low rate or did not produce them at all. No significant differences were found by studying the biomass, mycelium and sporeforming organs of the parent cultures and their mutants. However, their resistance to UV was not the same. Addition of the pigment apsergillin, isolated from the conidia of Aspergillus niger, to a suspension of the pigmentless (mutant) conidia of Penicillium notatum, the spores of Circinella muscae, and the vegetative cells of Micrococcus albus, before their irradiation with UV, considerably increased their resistance to this factor.     

Microorganisms of the upper layers of the atmosphere

Samples for microbiological studies were taken at a high altitude using meterological rockets with specially constructed analyzers so that no extraneous microorganisms could get into them before sampling. The analyzers were sterilized by gamma-irradiation before sampling. For the first time, microscopic fungi, bacteria and mycobacteria were detected at a height of 48--77 km. Five among six of the microbial species were found to contain pigments. The prevalence of these forms at high altitudes suggests that natural selection may take place in the upper atmospheric layer since cells containing chromogenic pigments (carotenoids, melanins) are more resistant to the action of ultraviolet rays.     

Isolation and characterization of melanin pigment from <Emphasis Type="Italic">Pleurotus cystidiosus</Emphasis> (telomorph of <Emphasis Type="Italic">Antromycopsis</Emphasis><Emphasis Type="Italic">macrocarpa</Emphasis>)

Melanins are enigmatic pigments that are produced by a wide variety of microorganisms including several species of bacteria and fungi. For more than 40 years, fungi have been known to produce pigments called melanins. Melanin pigment production by mushrooms was not intensively studied. The present study was carried out on isolation and characterization of melanin from an edible mushroom Pleurotus cystidiosus var. formosensis. The mushroom produced dark mucous mass of hyaline arthrospores on mycelium. The coremia exclusively produced dikaryotic arthrospores with the remnant of a clamp connection. Continuous cell extension and division in the coremium stipe supplied cells for arthroconidiation at the coremium apex, which is surrounded by a liquid droplet (coremioliquid). The black coloured coremea (conidia) were produced by Antromycopsis macrocarpa (anamorph of P. cystidiosus) when cultured on potato dextrose agar medium. The agar plate was incubated at continuous light illumination for high amount of pigment (coremea) production. The slimy layer of the coremea was extracted and partially purified by alkaline and acid treatment. The black pigment was confirmed as melanin based on UV, IR and EPR spectra apart from chemical analysis. This is the first report on characterization of melanin obtained from Pleurotus cystidiosus var. formosensis.     

A novel enzyme immunoassay for conidia of Fusarium oxysporum f.sp. cucumerinum F504

This study was undertaken to develop a new tool to study fusarial diseases of plants. Micro- and macro-conidia of a strain (F504) of Fusarium oxysporum were isolated and antiserum against the conidia was elicited in rabbits. A highly specific and sensitive competitive-type enzyme-linked immunosorbent assay (ELISA) for conidia of the strain was developed using the antiserum with beta-D-galactosidase-labeled anti-rabbit IgG as the secondary antibody and conidia fragments of the strain as antigen attached to Amino-Dylark solid-phase balls. The assay was highly specific to conidia of the strain F504, while conidia-free hypha of the strain F504 as well as all other microorganisms tested including nine other strains of Fusarium species showed little cross-reactivity. Application of the ELISA to following the growth rates of conidia in hyphae of the strain F504 under several conditions are also reported.     

Structure and function of the conidiospore pigments of Penicillium cyclopium.

The cell wall of mature, green condiospores of Penicillium cyclopium Westling contains at least two pigments: A green chromoprotein which is extractable by means of formic acid or liquified phenol and a black insoluble pigment. Both fractions after long term treatment with boiling conc. HCl leave black amorphous residues which, due to their chemical and physico-chemical properties, belong to the group of melanins. The chemical structure of these melanins is still unidentified. No degradation products typical for indol-type or catechol-type melanins have so far been detected. During spore maturation parallel to an increase of pigmentation (determined by remission), the melanin residue left after acid hydrolysis of spores increases. The mature, dark green spores of the wild type strain contain about 40% melanin, the yellow-green spores of the mutant aux-glu 1 about 36%. The unpigmented spores of mutant res-eth 1 possess a melanin content of only about 5%. This value is nearly the same as that found in hyphae, which in all strains are yellowish-brown. The heavily pigmented condia of the wild type strain are about 100-times less sensitive to UV-radiation compared with the unpigmented spores of the mutant res-eth 1. The reduced sensitivity indicates that, as with other microorganisms, the conidia pigments of P. cyclopium are protective components of the spores.     

Monascus secondary metabolites: production and biological activity

The genus Monascus, comprising nine species, can reproduce either vegetatively with filaments and conidia or sexually by the formation of ascospores. The most well-known species of genus Monascus, namely, M. purpureus, M. ruber and M. pilosus, are often used for rice fermentation to produce red yeast rice, a special product used either for food coloring or as a food supplement with positive effects on human health. The colored appearance (red, orange or yellow) of Monascus-fermented substrates is produced by a mixture of oligoketide pigments that are synthesized by a combination of polyketide and fatty acid synthases. The major pigments consist of pairs of yellow (ankaflavin and monascin), orange (rubropunctatin and monascorubrin) and red (rubropunctamine and monascorubramine) compounds; however, more than 20 other colored products have recently been isolated from fermented rice or culture media. In addition to pigments, a group of monacolin substances and the mycotoxin citrinin can be produced by Monascus. Various non-specific biological activities (antimicrobial, antitumor, immunomodulative and others) of these pigmented compounds are, at least partly, ascribed to their reaction with amino group-containing compounds, i.e. amino acids, proteins or nucleic acids. Monacolins, in the form of β-hydroxy acids, inhibit hydroxymethylglutaryl-coenzyme A reductase, a key enzyme in cholesterol biosynthesis in animals and humans.     

PCR-RFLP is used to investigate relations among species in the entomopathogenic genera Eryniopsis and Entomophaga

The shape and nucleation of primary conidia are important characters in the classification of the Entomophthoraceae (Zygomycetes). The five species in the genus Eryniopsis vary in the shapes of primary conidia, although within most genera in the order Entomophthorales species have the same shapes of primary conidia. Using PCR-RFLP, we investigated two species in Eryniopsis, Ery. caroliniana with oblong-ovoid primary conidia and Ery. ptychopterae with pear-shaped primary conidia, with five species of Entomophaga, all having pear-shaped conidia. Molecular results merged with morphological data indicate that Ery. ptychopterae belongs in the genus Entomophaga while Ery. caroliniana clearly differs from Entomophaga. Ery. ptychopterae and Ery. transitans are transferred to the genus Entomophaga. Our results support the idea that morphology of primary conidia is of major importance in defining entomophthoralean genera. These results also show that such studies can be conducted with species that have not been isolated, if fungal-filled cadavers can be obtained.     

Use of the i-STAT portable clinical analyzer in mice

The use of portable chemistry analyzers is an attractive option for obtaining clinical pathology panels in mice, because these analyzers require only small volumes of whole blood. However, in studies with other animals, portable analyzers do not always agree with results obtained using standard laboratory equipment. The authors evaluated the use of the i-STAT handheld portable clinical analyzer compared to the use of standard nonportable laboratory instruments in mice. As shown with other species, the i-STAT results did not always agree with standard laboratory instruments; however, the i-STAT does show reliability for certain chemistry assays.     

The nitrosophenol system of iron metabolism regulation in actinomycetes

Iron-containing o-nitrosophenol compounds have been isolated from 64 strains of microorganisms of 18 species. The analysis of author's and published data allowed to divide iron-containing microbial compounds into two physiological groups: ferri-ionophores, i. e. compounds participating in active iron transfer, and those immobilizing iron at increase in its content in medium up to 10(-4)-10(-2) M. O-nitrosophenol pigments were found to belong to the latter group. A scheme of participation of o-nitrosophenol compounds in regulation of iron metabolism in actinomycetes has been proposed.     

Scanning electron microscopy of Penicillium conidia

The morphology of conidia in 211 species and 12 varieties belonging to the genus Penicillium Link ex Gray have been studied and compared.According to surface ornamentation, conidia have been classified into six groups: A, smooth-walled (7% of the species); B, delicately roughened (13%); C, warty (28%); D, echinate (10%); E, striate with low irregular ridges (36%); and F, striate with scarce high ridges or bars (6%). Whereas the first two groups are closely related in both shape and average size, a gradual reduction was observed in size and in the length/width (l/w) ratio in the remaining groups. Echinate conidia were globose, having the largest average size. Only four species produced conidia not surpassing 2 m in diameter. Maximum length observed was 8 m, and most elongated conidia had a l/w ratio of 3.5. Forty per cent of the species studied had globose conidia.Conidia of the monoverticillate species were generally smaller, more globose and frequently with ridges. In the Asymmetrica, the conidia were generally larger, and showed ridges in comparatively few species. Conidia of the Symmetrica, which were frequently striate with ridges, presented the most elongated forms. The largest average size was found in the conidia of the Polyverticillata which were generally warty. Finally, we have considered the variations in surface ornamentation of conidia during the evolution of the genus Penicillium and drawn attention to their possible relationship with certain habitats and ways of conidial dispersion.     

Kinetic studies on the aggregation of Aspergillus niger conidia

Morphology has a crucial effect on productivity and the supply of substrate for cultures of filamentous fungi. However, cultivation parameters leading to the desired morphology are often chosen empirically as the mechanisms governing the processes involved are usually unknown. For coagulating microorganisms like Aspergillus niger the morphological development is considered to start with the aggregation of conidia right after inoculation. To elucidate the mechanism of this process, kinetic studies were carried out using an in-line particle size analyzer. Based on the data obtained from these experiments a model for conidial aggregation is proposed in this article. It consists of two separate aggregation steps. The first one takes place immediately after inoculation, but only leads to a small decrease of total particle concentration. Most suspended conidia aggregate after a second aggregation step triggered by germination and hyphal growth. Aggregation velocity of this second phase is linearly dependent on the particle growth rate.     

Anamorphs in Mycocalicium albonigrum and M. subtile (Caliciales)

Anamorphs of Mycocalicium albonigrum and M. subtile have been obtained in culture from ascospores, conidia collected in the field, and conidia produced in culture. The ontogenies of the conidiomata and conidia are described. Differences between the species with respect to cultural characteristics, septation of ascospores during germination, occurrence of chlamydospores and pyriform cells as well as ultrastructural details of conidiogenous cells and conidia were studied. Conidia are suggested to possibly be important in the dispersal of the species.     

The contribution of melanin to microbial pathogenesis

Melanins are enigmatic pigments that are produced by a wide variety of microorganisms including several species of pathogenic bacteria, fungi and helminths. The study of melanin is difficult because these pigments defy complete biochemical and structural analysis. Nevertheless, the availability of new reagents in the form of monoclonal antibodies and melanin-binding peptides, combined with the application of various physical techniques, has provided insights into the process of melanization. Melanization is important in microbial pathogenesis because it has been associated with virulence in many microorganisms. Melanin appears to contribute to virulence by reducing the susceptibility of melanized microbes to host defence mechanisms. However, the interaction of melanized microbes and the host is complex and includes immune responses to melanin-related antigens. Production of melanin has also been linked to protection against environmental insults. Interference with melanization is a potential strategy for antimicrobial drug and pesticide development. The process of melanization poses fascinating problems in cell biology and provides a type of pathogenic strategy that is common to highly diverse pathogens.     

微生物产色素机制及其生物活性

近年来,随着人工合成色素的大量使用引起一系列环境和健康问题,增加了人们对安全、无毒天然色素的需求.天然色素主要来源于植物和微生物,由于植物生长周期较长使植物源色素在大规模应用中受限.与植物源天然色素相比,微生物源色素易于大规模快速培养,具有更广阔的应用前景.本文系统总结了不同微生物源色素产生机制,及其在抗菌、抗氧化及抗...     

Yeast and yeast-like fungi associated with dry indehiscent fruits of Nothofagus nervosa in Patagonia, Argentina

Nothofagus nervosa (Raulí) is a native tree species that yields valuable timber. It was overexploited in the past and is currently included in domestication and conservation programs. Several research programs have focused on the characterization of epiphytic microorganisms because it has been demonstrated that they can affect plant-pathogen interactions and/or promote plant growth. Although the microbial ecology of leaves has been well studied, less is known about microorganisms occurring on seeds and noncommercial fruits. In this work, we analyzed the yeast and yeast-like fungi present on N.?nervosa fruits destined for the propagation of this species, as well as the effects of fruit preservation and seed dormancy-breaking processes on fungal diversity. Morphological and molecular methods were used, and differences between fungal communities were analyzed using a similarity index. A total of 171 isolates corresponding to 17 species were recovered, most of which belong to the phylum Ascomycota. The majority of the species develop mycelia, produce pigments and mycosporines, and these adaptation strategies are discussed. It was observed that the preservation process considerably reduced yeast and yeast-like fungal diversity. This is the first study concerning microbial communities associated with this ecologically and economically important species, and the information presented is relevant to domestication programs.     

Cell culture monitoring via an auto‐sampler and an integrated multi‐functional off‐line analyzer

Mammalian cell‐based bioprocesses are used extensively for production of therapeutic proteins. Off‐line monitoring of such cultivations via manual sampling is often labor‐intensive and can introduce operator‐dependent error into the process. An integrated multi‐functional off‐line analyzer, the BioProfile FLEX (NOVA Biomedical, Waltham MA) has been developed, which combines the functionality of three off‐line analyzers (a cell counter, an osmometer, and a gas/electrolyte & nutrient/metabolite bio‐profile analyzer) into one device. In addition, a novel automated sampling system has also been developed that allows the BioProfile FLEX to automatically analyze the culture conditions in as many as ten bioreactors. This is the first report on the development and function of this integrated analyzer and an auto‐sampler prototype for monitoring of mammalian cell cultures. Evaluation of the BioProfile FLEX was conducted in two separate laboratories and involved two BioProfile FLEX analyzers and two sets of reference analyzers (Nova BioProfile 400, Beckman‐Coulter Vi‐Cell AS, and Advanced Instruments Osmometer 3900), 13 CHO cell lines and over 20 operators. In general, BioProfile FLEX measurements were equivalent to those obtained using reference analyzers, and the auto‐sampler did not alter the samples it provided to the BioProfile FLEX. These results suggest that the system has the potential to dramatically reduce the manual labor involved in monitoring mammalian cell bioprocesses without altering the quality of the data obtained, and integration with a bioreactor control system will allow feedback control of parameters previously available only for off‐line monitoring. © 2009 American Institute of Chemical Engineers Biotechnol. Prog., 2010     

靛蓝及其同类色素的微生物生产与转化

靛蓝类色素广泛应用于印染、食品和医药工业, 其环境友好的合成或生产途径越来越受到人们的关注, 特别是微生物生物合成。已经鉴定和分离了能够合成靛蓝类色素的多种微生物, 并且明确了起催化作用的主要是单加氧酶和双加氧酶。已经克隆和利用了一些加氧酶的基因, 构建了工程菌, 优化了其发酵过程。同时, 微生物合成靛蓝的生物转化也已经起步。这些进展将带来环境友好的靛蓝类色素的合成与生产。     

Statistical and molecular analyses of evolutionary significance of red-green color vision and color blindness in vertebrates

Red-green color vision is strongly suspected to enhance the survival of its possessors. Despite being red-green color blind, however, many species have successfully competed in nature, which brings into question the evolutionary advantage of achieving red-green color vision. Here, we propose a new method of identifying positive selection at individual amino acid sites with the premise that if positive Darwinian selection has driven the evolution of the protein under consideration, then it should be found mostly at the branches in the phylogenetic tree where its function had changed. The statistical and molecular methods have been applied to 29 visual pigments with the wavelengths of maximal absorption at approximately 510-540 nm (green- or middle wavelength-sensitive [MWS] pigments) and at approximately 560 nm (red- or long wavelength-sensitive [LWS] pigments), which are sampled from a diverse range of vertebrate species. The results show that the MWS pigments are positively selected through amino acid replacements S180A, Y277F, and T285A and that the LWS pigments have been subjected to strong evolutionary conservation. The fact that these positively selected M/LWS pigments are found not only in animals with red-green color vision but also in those with red-green color blindness strongly suggests that both red-green color vision and color blindness have undergone adaptive evolution independently in different species.     

Antioxidant capacity of novel pigments from an Antarctic bacterium

In Antarctica microorganisms are exposed to several conditions that trigger the generation of reactive oxygen species, such as high UV radiation. Under these conditions they must have an important antioxidant defense system in order to prevent oxidative damage. One of these defenses are pigments which are part of the non-enzymatic antioxidant mechanisms. In this work we focused on the antioxidant capacity of pigments from an Antarctic microorganism belonging to Pedobacter genus. This microorganism produces different types of pigments which belong to the carotenoids group. The antioxidant capacity of a mix of pigments was analyzed by three different methods: 1,1-diphenyl-2-picrylhydrazyl, ROS detection and oxygen electrode. The results obtained from these approaches indicate that the mix of pigments has a strong antioxidant capacity. The oxidative damage induced by UVB exposure to liposomes was also analyzed. Intercalated pigments within the liposomes improved its resistance to lipid peroxidation. Based on the analysis carried out along this research we conclude that the antioxidant properties of the mix of pigments protect this bacterium against oxidative damage. These properties make this mix of pigments a powerful antioxidant mixture with potential biotechnological applications.