Different Scenarios for the Origin and the Subsequent Succession of a Hypothetical Microbial Community in the Cloud Layer of Venus.

The possible existence of a microbial community in the venusian clouds is one of the most intriguing hypotheses in modern astrobiology. Such a community must be characterized by a high survivability potential under severe environmental conditions, the most extreme of which are very low pH levels and water activity. Considering different scenarios for the origin of life and geological history of our planet, a few of these scenarios are discussed in the context of the origin of hypothetical microbial life within the venusian cloud layer. The existence of liquid water on the surface of ancient Venus is one of the key outstanding questions influencing this possibility. We link the inherent attributes of microbial life as we know it that favor the persistence of life in such an environment and review the possible scenarios of life's origin and its evolution under a strong greenhouse effect and loss of water on Venus. We also propose a roadmap and describe a novel methodological approach for astrobiological research in the framework of future missions to Venus with the intent to reveal whether life exists today on the planet.

Water-Sulfuric Acid Foam as a Possible Habitat for Hypothetical Microbial Community in the Cloud Layer of Venus.

The data available at the moment suggest that ancient Venus was covered by extensive bodies of water which could harbor life. Later, however, the drastic overheating of the planet made the surface of Venus uninhabitable for Earth-type life forms. Nevertheless, hypothetical Venusian organisms could have gradually adapted to conditions within the cloud layer of Venus-the only niche containing liquid water where the Earth-type extremophiles could survive. Here we hypothesize that the unified internal volume of a microbial community habitat is represented by the heterophase liquid-gas foam structure of Venusian clouds. Such unity of internal space within foam water volume facilitates microbial cells movements and trophic interactions between microorganisms that creates favorable conditions for the effective development of a true microbial community. The stabilization of a foam heterophase structure can be provided by various surfactants including those synthesized by living cells and products released during cell lysis. Such a foam system could harbor a microbial community of different species of (poly)extremophilic microorganisms that are capable of photo- and chemosynthesis and may be closely integrated into aero-geochemical processes including the processes of high-temperature polymer synthesis on the planet's surface. Different complex nanostructures transferred to the cloud layers by convection flows could further contribute to the stabilization of heterophase liquid-gas foam structure and participate in chemical and photochemical reactions, thus supporting ecosystem stability.

Comparison of the Wild-Type Obligate Methylotrophic Bacterium Methylophilus quaylei and its Isogenic Streptomycin-Resistant Mutant via Metal Nanoparticle Generation.

Metal nanoparticles synthesized by green methods with the use of microorganisms are currently one of the most closely studied types of nanomaterials. It has accurately been shown that the characteristics of metal nanoparticles generated in the presence of different bacteria vary. For the two isogenic strains of obligate methylotrophic bacteria of the wild type (M. quaylei MTT) and its streptomycin-resistant mutant (M. quaylei SMR), the pleiotropic character of streptomycin resistance mutation in the SMR cells has been revealed. It has been shown that both cultures can generate silver nanoparticles. There is a dramatic difference in the formation of palladium nanoparticles, which are formed only in the presence of cells of the streptomycin-resistant mutant M. quaylei SMR. This study shows that closely related isogenic strains of obligate methylotrophic bacteria can be distinguished by the spectra of biogenic nanoparticles of two noble metals. While palladium nanoparticles are only generated by the cells of the streptomycin-resistant mutant M. quaylei SMR, biogenic silver nanoparticles can be generated from both cultures. Thus, the assessment of the ability of microorganisms to form biogenic nanoparticles of different metals allows the revelation of subtle metabolic differences of even close cultures.

Antimicrobial activity of silver salt and silver nanoparticles in different forms against microorganisms of different taxonomic groups.

The development of antiseptics and medical products (bandaging materials, sponges, etc.) based on silver nanoparticles is an essential task due to the growing resistance of pathogenic microorganisms to medicines long used in clinical practice. Using silver nanoparticles for the same purpose is promising, but the potential hazards and cumulative effects in the application of nanoparticles requires a thorough study of those materials. To evaluate the efficiency of antiseptics and medical products based on silver nanoparticles, it is necessary to conduct an in-depth study of the activity of silver nanoparticles in different forms and immobilized in carriers. The study examines the resistance of bacterial and fungal cultures to silver nanoparticles produced by chemical reduction and microbiological synthesis. The study of resistance was carried out in different growth phases of pathogenic microorganisms and in both liquid and solid media. Chemically and microbiologically synthesized nanoparticles were added in the form of a suspension, as well as encapsulated in chitosan-PVA matrices. It was experimentally discovered that, depending on the medium and form of the silver, the antibacterial effect would significantly differ due to changes in the mechanisms regarding the release of nanoparticles and their activity against the cells of pathogenic and potentially pathogenic microorganisms.

Halobacterium salinarum storage and rehydration after spray drying and optimization of the processes for preservation of carotenoids.

Spray drying is appropriate for the preservation of halophilic microorganisms due to the nature of these microorganisms, as they survive in adverse environmental conditions by being encapsulated in salt crystals. Artificial neural networks were in this study used to optimize practically significant spray-drying regimes of the C50-carotenoids producer Halobacterium salinarum. Immediately after drying, the samples contained up to 54% halobacterial biomass and less than 5% moisture, and the level of preservation of carotenoids was 95-97%. The storage of biomass at 4 °C resulted in the gradual degradation of the carotenoids, which reached 58-64% in the best samples after 1 year. A comprehensive study of changes in halobacteria biomass after spray drying and the nature of the damage provided new data on the survival and preservation of cells and biologically active substances in the various spray-drying regimes and at different storage times.

Efficient continuous biosynthesis of silver nanoparticles by activated sludge micromycetes with enhanced tolerance to metal ion toxicity.

The method for producing AgNPs by granules of activated sludge micromycetes with enhanced tolerance to metal ion toxicity - Penicillium glabrum, Fusarium nivale and Fusarium oxysporum has been developed; the optimum conditions for AgNP biosynthesis being found: the Ag+ ion concentration, duration of the contact of microbial cells with silver ions, a growth phase of microorganisms, medium composition, a рН value, mixing conditions, and also lighting intensity. The effect of Cl-, SO42- and HPO42- ions binding Ag+ ions was eliminated, that brought to significant increase of the yield of NPs. Under batch conditions, silver particles of 60-110 nanometers in size were formed with a 65% yield. It was established that the nanoparticles were covered with microbial cell membrane proteins composed up to 70% by weight of the NPs that prevented their aggregation. In addition, it was the first time stable AgNPs had been formed by continuous AgNP biosynthesis by living cells of F. oxysporum with an 80% yield for a long time.

High-effective cultivation of Halobacterium salinarum providing with bacteriorhodopsin production under controlled stress.

Submerged growth of Halobacterium salinarum and therefore synthesis of bacteriorhodopsin (BR) and carotenoids depend greatly on products of both chemical and/or photochemical oxidation of medium components and cellular metabolism which act as inhibitors. Some cultivation variants which allowed eliminating an adverse effect of inhibitors on biomass accumulation and BR synthesis are reviewed. The application of activated charcoal or ion exchange resin as adsorbents at preparing inoculums and the main cultivation stages was shown to allow controlling, namely lowering overstress of the halobacterial cells by metabolites. The halobacterial biomass containing BR up to 1,750mgL(-1) and the minimum amount of carotinoids that would BR greatly facilitate isolation was accumulated up to 45gL(-1) during eight-day cultivation with cell recycling through adsorbent suspension in a fed-batch mode. To control BR biosynthesis the express method of BR quantification based on colour shades of cell suspension was developed.

Toxic influence of silver and uranium salts on activated sludge of wastewater treatment plants and synthetic activated sludge associates modeled on its pure cultures.

Toxic impact of silver and uranium salts on activated sludge of wastewater treatment facilities has been studied. Some dominating cultures (an active nitrogen fixer Agrobacterium tumifaciens (A.t) and micromyces such as Fusarium nivale, Fusarium oxysporum, and Penicillium glabrum) have been isolated and identified as a result of selection of the activated sludge microorganisms being steadiest under stressful conditions. For these cultures, the lethal doses of silver amounted 1, 600, 50, and 300 µg/l and the lethal doses of uranium were 120, 1,500, 1,000, and 1,000 mg/l, respectively. A.tumifaciens is shown to be more sensitive to heavy metals than micromyces. Synthetic granular activated sludge was formed on the basis of three cultures of the isolated micromyces steadiest against stress. Its granules were much more resistant to silver than the whole native activated sludge was. The concentration of silver causing 50 % inhibition of synthetic granular activated sludge growth reached 160-170 µg/l as far as for the native activated sludge it came only to 100-110 µg/l.

Antiamoebin I in methanol solution: rapid exchange between right-handed and left-handed 3(10)-helical conformations.

Antiamoebin I (Aam-I) is a membrane-active peptaibol antibiotic isolated from fungal species belonging to the genera Cephalosporium, Emericellopsis, Gliocladium, and Stilbella. Antiamoebin I has the amino acid sequence: Ac-Phe(1)-Aib-Aib-Aib-Iva-Gly-Leu-Aib(8)-Aib-Hyp-Gln-Iva-Hyp-Aib-Pro-Phl(16). By using the uniformly (13)C,(15)N-labeled sample of Aam-I, the set of conformationally dependent J couplings and (3h)J(NC) couplings through H-bonds were measured. Analysis of these data along with the data on magnetic nonequivalence of the (13)C(beta) nuclei (Deltadelta((13)C(beta))) in Aib and Iva residues allowed us to draw the univocal conclusion that the N-terminal part (Phe(1)-Gly(6)) of Aam-I in MeOH solution is in fast exchange between the right-handed and left-handed 3(10)-helical conformations, with an approximately equal population of both states. An additional conformational exchange process was found at the Aib(8) residue. The (15)N-NMR-relaxation and CD-spectroscopy measurements confirmed these findings. Molecular modeling and Monte Carlo simulations revealed that both exchange processes are correlated and coupled with significant hinge-bending motions around the Aib(8) residue. Our results explain relatively low activity of Aam-I with respect to other 15-amino acid residue peptaibols (for example, zervamicin) in functional and biological tests. The high dynamic 'propensity' possibly prevents both initial binding of the antiamoebin to the membrane and subsequent formation of stable ionic channels according to the barrel-stave mechanism.

Biosynthetic uniform 13C,15N-labelling of zervamicin IIB. Complete 13C and 15N NMR assignment.

Zervamicin IIB is a member of the alpha-aminoisobutyric acid containing peptaibol antibiotics. A new procedure for the biosynthetic preparation of the uniformly 13C- and 15N-enriched peptaibol is described This compound was isolated from the biomass of the fungus-producer Emericellopsis salmosynnemata strain 336 IMI 58330 obtained upon cultivation in the totally 13C, 15N-labelled complete medium. To prepare such a medium the autolysed biomass and the exopolysaccharides of the obligate methylotrophic bacterium Methylobacillus flagellatus KT were used. This microorganism was grown in totally 13C, 15N-labelled minimal medium containing 13C-methanol and 15N-ammonium chloride as the only carbon and nitrogen sources. Preliminary NMR spectroscopic analysis indicated a high extent of isotope incorporation (> 90%) and led to the complete 13C- and 15N-NMR assignment including the stereospecific assignment of Aib residues methyl groups. The observed pattern of the structurally important secondary chemical shifts of 1H(alpha), 13C=O and 13C(alpha) agrees well with the previously determined structure of zervamicin IIB in methanol solution.