RESUMO
Screening of alkane hydroxylase genes (alkB) was performed in the thermophilic aerobic bacteria of the genus Geobacillus. Total DNA was extracted from the biomass of 11 strains grown on the mixture of saturated C10-C20 hydrocarbons, PCR amplification of fragments of alkB genes was performed with degenerate oligonucleotide primers, PCR products were cloned and sequenced. For the first time in the genome of thermophilic bacteria the presence of a set of alkB gene homologues was revealed. The strains each contain three to six homologues among which only two are universal for all of the strains. Comparative phylogenetic analysis of the nucleotide sequences and the inferred amino acid sequences showed close relatedness of six of the revealed variants of geobacilli sequences to the alkB4, alkB3, and alkB2 genes that had previously been revealed by other authors in Rhodococcus erythropolis strains NRRL B-16531 and Q15. The rest two variants of alkB sequences were unique. Analysis of the GC composition of all the Geobacillus alkB homologues revealed closer proximity to the rhodococcal chromosomal DNA than to the chromosomal DNA of geobacilli. This may be an indication of the introduction of the alkB genes into the Geobacillus genome by interspecies horizontal transfer; and rhodococci or other representatives of the Actinobacteria phylum were probably the donors of these genes. Analysis of the codon usage in fragments of alkB genes confirms the suggestion that the pool of these genes is common to the majority of Gram-positive and certain Gram-negative bacteria. Formation of a set of several alkB homologues in a genome of a particular microorganism may result from free gene exchange within this pool.
Assuntos
Bacillaceae/genética , Proteínas de Bactérias/genética , Citocromo P-450 CYP4A/genética , Transferência Genética Horizontal/fisiologia , Genes Bacterianos/fisiologia , Filogenia , Sequência de Aminoácidos/genética , Bacillaceae/enzimologia , Bacillaceae/crescimento & desenvolvimento , Bacillaceae/isolamento & purificação , Proteínas de Bactérias/metabolismo , Biomassa , Citocromo P-450 CYP4A/metabolismo , Rhodococcus/enzimologia , Rhodococcus/genética , Análise de Sequência de Proteína , Homologia de Sequência de AminoácidosRESUMO
Water-electrolyte homeostasis and its basic regulatory hormones have been studied in 36 patients with neurosurgical brain pathology during surgical interventions performed under balanced anesthesia (sodium hydroxybutyrate combined with NLA drugs). The study has revealed 3 types of reactions in hormones regulating water-electrolyte homeostasis: type I--a decrease in vasopressin (VP) concentration and renin-angiotensin-aldosterone system (RAAS) activity; type II--VP increase and RAAS activation; type III--an increase in VP content and RAAS disbalance. It has been shown that type I reaction is accompanied by marked osmotic disturbances. The impact of sodium hydroxybutyrate on RAAS is manifested in aldosterone secretion suppression.
Assuntos
Anestesia , Lesões Encefálicas/cirurgia , Neoplasias Encefálicas/cirurgia , Oxibato de Sódio , Equilíbrio Hidroeletrolítico/fisiologia , Adolescente , Adulto , Lesões Encefálicas/fisiopatologia , Neoplasias Encefálicas/fisiopatologia , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , NeuroleptanalgesiaAssuntos
Alcanos/metabolismo , Regulação Bacteriana da Expressão Gênica , Geobacillus/crescimento & desenvolvimento , Geobacillus/genética , Oxigenases de Função Mista/genética , Biodegradação Ambiental , Metabolismo dos Carboidratos/genética , Geobacillus/enzimologia , Petróleo , RNA Mensageiro/genética , Homologia de Sequência do Ácido Nucleico , Transcrição GênicaRESUMO
The number of microorganisms of major metabolic groups and the rates of sulfate-reducing and methanogenic processes in the formation waters of the high-temperature horizons of Dagang oilfield have been determined. Using cultural methods, it was shown that the microbial community contained aerobic bacteria oxidizing crude oil, anaerobic fermentative bacteria, sulfate-reducing bacteria, and methanogenic bacteria. Using cultural methods, the possibility of methane production from a mixture of hydrogen and carbon dioxide (H2 + CO2) and from acetate was established, and this result was confirmed by radioassays involving NaH14CO3 and 14CH3COONa. Analysis of 16S rDNA of enrichment cultures of methanogens demonstrated that these microorganisms belong to Methanothermobacter sp. (M. thermoautotrophicus), which consumes hydrogen and carbon dioxide as basic substrates. The genes of acetate-utilizing bacteria were not identified. Phylotypes of the representatives of Thermococcus spp. were found among 16S rDNAs of archaea. 16S rRNA genes of bacterial clones belong to the orders Thermoanaerobacteriales (Thermoanaerobacter, Thermovenabulum, Thermacetogenium, and Coprothermobacter spp.), Thermotogales, Nitrospirales (Thermodesulfovibrio sp.) and Planctomycetales. 16S rDNA of a bacterium capable of oxidizing acetate in the course of syntrophic growth with H2-utilizing methanogens was found at high-temperature petroleum reservoirs for the first time. These results provide further insight into the composition of microbial communities of high-temperature petroleum reservoirs, indicating that syntrophic processes play an important part in acetate degradation accompanied by methane production.
Assuntos
Euryarchaeota/genética , Petróleo/microbiologia , Filogenia , RNA Ribossômico 16S/genética , Microbiologia do Solo , Anaerobiose/fisiologia , China , Euryarchaeota/crescimento & desenvolvimento , Temperatura Alta , Análise de Sequência de RNA/métodosRESUMO
The distribution and species diversity of aerobic organotrophic bacteria in the Dagan high-temperature oil field (China), which is exploited via flooding, have been studied. Twenty-two strains of the most characteristic thermophilic and mesophilic aerobic organotrophic bacteria have been isolated from the oil stratum. It has been found that, in a laboratory, the mesophilic and thermophilic isolates grow in the temperature, pH, and salinity ranges characteristic of the injection well near-bottom zones or of the oil stratum, respectively, and assimilate a wide range of hydrocarbons, fatty acids, lower alcohols, and crude oil, thus exhibiting adaptation to the environment. Using comparative phylogenetic 16S rRNA analysis, the taxonomic affiliation of the isolates has been established. The aerobic microbial community includes gram-positive bacteria with a high and low G+C content of DNA, and gamma and beta subclasses of Proteobacteria. The thermophilic bacteria belong to the genera Geobacillus and Thermoactinomyces, and the mesophilic strains belong to the genera Bacillus, Micrococcus, Cellulomonas, Pseudomonas, and Acinetobacter. The microbial community of the oil stratum is dominated by known species of the genus Geobacillus (G. subterraneus, G. stearothermophilus, and G. thermoglucosidasius) and a novel species "Geobacillus jurassicus." A number of novel thermophilic oil-oxidizing bacilli have been isolated.