[Regulation of geochemical activity of microorganisms in a petroleum reservoir by injection of H2O2 or water-air mixture].

In the course of pilot trials of biotechnologies for the enhancement of oil recovery in the Gangxi bed of the Dagang oil field (China), microbiological processes were investigated. The biotechnologies were based on injection into the petroleum reservoir of different oxygen sources (H2O2 solution or a water-air mixture) with nitrogen and phosphorus salts. The injection of water-air mixture with nitrogen and phosphorus salts resulted in an increase in the number of aerobic and anaerobic organotrophic bacteria, rates of sulfate reduction and methanogenesis in formation water and also the content of CO2 (from 4.8-12 to 15-23.2%) and methane (from 86-88 to 91.8%) in the gas. The preferential consumption of isotopically light bicarbonate by methanogens resulted in a higher content of the light 12C in methane; the delta13C/CH4 value changed from -45.1...-48.3 to -50.7...-59.3 per thousand). At the same time, mineral carbonates of the formation water became isotopically heavier; the delta13C/Sigmacarbonates value increased from 3.4...4.0 to 5.4...9.6 per thousand. Growth of hydrocarbon-oxidizing bacteria was accompanied by production of biosurfactants and decreased interfacial tension of formation water. Injection of H2O2 solution resulted in the activation of aerobic processes and in suppression of both sulfate reduction and methanogenesis. Methane content in the gas decreased from 86-88 to 75.4-79.8%, probably due to its consumption by methanotrophs. Due to consumption of isotopically light methane, the residual methane carbon became heavier, with the delta13C/CH4 values from -39.0 to -44.3 per thousand. At the same time, mineral carbonates of the formation water became isotopically considerably lighter; the delta13C/Sigmacarbonates value decreased from 5.4... 9.6 to -1.4...2.7 per thousand). The additional amount of oil recovered during the trial of both variants of biotechnological treatment was 3819 t.

[Microbiological and production characteristics of the high-temperature Kongdian bed revealed during field trial of biotechnology for the enhancement of oil recovery].

Microbiological technology for the enhancement of oil recovery based on the activation of the stratal microflora was tested in the high-temperature horizons of the Kongdian bed (60 degrees C) of the Dagang oil field (China). This biotechnology consists in the pumping of a water-air mixture and nitrogen and phosphorus mineral salts into the oil stratum through injection wells in order to stimulate the activity of the stratal microflora which produce oil-releasing metabolites. Monitoring of the physicochemical, microbiological, and production characteristics of the test site has revealed large changes in the ecosystem as a result of the application of biotechnology. The cell numbers of thermophilic hydrocarbon-oxidizing, fermentative, sulfate-reducing, and methanogenic microorganisms increased 10-10 000-fold. The rates of methanogenesis and sulfate reduction increased in the near-bottom zone of the injection wells and of some production wells. The microbial oil transformation was accompanied by the accumulation of bicarbonate ions, volatile fatty acids, and biosurfactants in the formation waters, as well as of CH4 and CO2 both in the gas phase and in the oil. Microbial metabolites promoted the additional recovery of oil. As a result of the application of biotechnology, the water content in the production liquid from the test site decreased, and the oil content increased. This allowed the recovery of more than 14000 tons of additional oil over 3.5 years.

[Microbiological investigations of high-temperature horizons of the Kongdian petroleum reservoir in connection with field trial of a biotechnology for enhancement of oil recovery].

The physicochemical conditions and microbiological characteristics of the formation waters of the Kongdian bed of the Dagang oil field (China) were studied. It was demonstrated that this bed is a high-temperature ecosystem with formation waters characterized by low mineralization. The concentrations of nitrogen and phosphorus compounds, as well as of electron acceptors, are low. Oil and oil gas are the main organic matter sources. The bed is exploited with water-flooding. The oil stratum was inhabited mostly by anaerobic thermophilic microorganisms, including fermentative (10(2)-10(5) cells/ml), sulfate-reducing (0-10(2) cells/ml), and methanogenic (0-10(3) cells/ml) microorganisms. Aerobic bacteria were detected mainly in the near-bottom zone of injection wells. The rate of sulfate reduction varied from 0.002 to 18.940 microg S(2-) l(-1) day(-1) and the rate of methanogenesis from 0.012 to 16.235 microg CH4 l(-1) day(-1). Microorganisms with great biotechnological potential inhabited the bed. Aerobic thermophilic bacteria were capable of oxidizing oil with the formation of biomass, the products of partial oxidation of oil (volatile acids), and surfactants. During growth on the culture liquid of oiloxidizing bacteria, methanogenic communities produced methane and carbon dioxide, which also had oil-releasing capabilities. Using various labeled tracers, the primary filtration flows of injected solutions at the testing site were studied. Our comprehensive investigations allowed us to conclude that the tested method for microbial enhancement of oil recovery based on the activation of the stratal microflora can be applied in the Kongdian bed horizons.

[Distribution and activity of microorganisms in the deep repository for liquid radioactive waste at the Siberian Chemical Combine].

The physicochemical conditions, composition of microbial communities, and the rates of anaerobic processes in the deep sandy horizons used as a repository for liquid radioactive wastes (LRW) at the Siberian Chemical Combine (Seversk, Tomsk oblast), were studied. Formation waters from the observation wells drilled into the production horizons of the radioactive waste disposal site were found to be inhabited by microorganisms of different physiological groups, including aerobic organotrophs, anaerobic fermentative, denitrifying, sulfate-reducing, and methanogenic bacteria. The density of microbial population, as determined by cultural methods, was low and usually did not exceed 10(4) cells/ml. Enrichment cultures of microorganisms producing gases (hydrogen, methane, carbon dioxide, and hydrogen sulfide) and capable of participation in the precipitation of metal sulfides were obtained from the waters of production horizons. The contemporary processes of sulfate reduction and methanogenesis were assayed; the rates of these terminal processes of organic matter destruction were found to be low. The denitrifying bacteria from the underground repository were capable of reducing the nitrates contained in the wastes, provided sources of energy and biogenic elements were available. Biosorption of radionuclides by the biomass of aerobic bacteria isolated from groundwater was demonstrated. The results obtained give us insight into the functional structure of the microbial community inhabiting the waters of repository production horizons. This study indicates that the numbers and activity of microbial cells are low both inside and outside the zone of radioactive waste dispersion, in spite of the long period of waste discharge.

[Degradation of machine oil by nocardiform bacteria]. / Degradatsiia neftianykh masel nokardiopodobnymi bakteriiami.

Gas liquid chromatography (GLC) was used for the first time to screen for machine oil-degrading microorganisms. Oil degradation was evaluated from the microorganism respiratory activity during the utilization of oil as the sole carbon and energy source. The results are consistent with those obtained by the conventional weighing method. Substrate specificity of the active strains with respect to different machine oils was studied. Bacterial communities exhibited the highest activity, whereas a Rhodococcus erythropolis strain was the most active among pure cultures. Various stages of bacterial interaction with oil drops were followed by means of fluorescent microscopy.

[Physiologo-biochemical properties of a strain of Beijerinckia mobilis 1phi Phn+--a degrader of polycyclic aromatic hydrocarbons]. / Fizilogo-biokhimicheskie svoistva shtamma Beijerinckia mobilis 1phi Phn+--destruktora politsiklicheskikh aromaticheskikh uglevodorodov.

Beijerinckia mobilis 1f capable of degrading polycyclic aromatic hydrocarbons (PAHs) was isolated from a soil contaminated with creosote. Strain 1f could utilize phenanthrene and naphthalene as the sole sources of carbon. The mean rate of phenanthrene degradation during culture growth was 7-8 micrograms/(ml h). After cultivation under nonselective conditions, strain 1f retained its ability to degrade phenanthrene. Cometabolism considerably widened the range of PAHs that could be transformed by strain 1f. The strain was able to grow in a mineral medium with creosote as the sole source of carbon. After 30 days of cultivation in this medium, the total concentration of PAHs decreased from 665.5 mg/l to 170 mg/l.

The catalytic mechanism of tyrosine phenol-lyase from Erwinia herbicola: the effect of substrate structure on pH-dependence of kinetic parameters in the reactions with ring-substituted tyrosines.

Apparently homogeneous tyrosine phenol-lyase (TPL) from Erwinia herbicola has been prepared by a new method. The pH-dependencies of the main kinetic parameters for the reactions of Erwinia TPL with tyrosine, 2-fluorotyrosine, 3-fluorotyrosine, 2-chlorotyrosine, and 3,4-dihydroxyphenylalanine (DOPA) have been studied. The pattern of pH-dependence of V(max) depends on the nature of the substituent in the aromatic ring. For the substrates bearing small substituents (H, 2-F, 3-F) V(max) values were found to be pH-independent. For 2-chlorotyrosine and DOPA V(max) decreased at lower pH, the effect being described by equation with one pKa. Generally two bases are reflected in the pH dependence of V(max)/Km. The first base, probably is responsible for the abstraction of alpha-proton, while the second one, interacts with the phenolic hydroxyl at the stage of binding. The reaction of TPL with DOPA differs from the reactions with other tyrosines by the requirement of an additional base which is reflected in the pH-profiles of both V(max) and V(max)/Km. For the reaction of TPL from Citrobacter intermedius with DOPA only V(max)/Km values could be determined. The activity of Citrobacter enzyme towards DOPA is considerably less than that of E. herbicola enzyme, and its maximal value is attained at higher pH.

[Monochlorophenols as enzyme substrates for the preparatory metabolism of phenol in Candida tropicalis yeasts]. / Monokhlorfenoly kak substraty fermentov podgotovitel'nogo metabolizma fenola u drozhzhei Candida tropicalis.

The object of this work was to find out whether Candida tropicalis can be used for monochlorophenol degradation. Phenol monooxygenase and pyrocatechase, enzymes involved in preparatory phenol metabolism were shown to catalyse transformation of 3- and 4-chlorophenols. Phenol monooxygenase catalyses hydroxylation of 3- and 4-chlorophenols to 4-chloropyrocatechol which yields beta-chloromuconic acid under the action of pyrocatechase. Synthesis of phenol monooxygenase is induced by 3- and 4-chlorophenols. beta-Chloromuconic acid is a terminal product of 3- and 4-chlorophenol transformation under neutral conditions. In a weakly acid medium (the Rieder medium, phosphate buffer, pH 5.5), transformation of these chlorophenols terminates with spontaneous lactonization of beta-chloromuconic acid and its dehalogenation. C. tropicalis hardly transforms 2-chlorophenol although certain oxygen uptake occurs in its presence. 3- and 4-chlorophenols are not nutrient sources for C. tropicalis. The yeast has not been adapted to 3- and 4-chlorophenols as sole nutrient sources.

[Preparatory metabolism of p-hydroxybenzoic acid in Candida tropicalis]. / Podgotovitel'nyi metabolizm para-oksibenzoinoi kisloty u drozhzhei Candida tropicalis.

A technique of experimental adaptation was used to obtain mutants of Candida tropicalis which were able to utilize p-hydroxybenzoic acid as the sole source of carbon and energy. The preparatory metabolism of p-hydroxybenzoic acid involves the following stages: PHBA leads to quinol leads to hydroxyquinol leads to maleylacetic acid leads to beta-ketoadipic acid. The enzyme system which catalyzes oxidative decarboxylation of PHBA mediates also oxidative decarboxylation of protocatechuic, beta-resorcylic and gallic acids, i.e. compounds having a hydroxyl group in para position with respect to the carboxyl of hydroxyl derivatives of benzoic acid. Benzoic, salicyclic and gentisic acids are not substrates of this enzyme system. A technique is proposed for rapid indentification of beta-ketoadipic acid by thin-layer chromatography. The authors believe that methods used to study preparatory metabolism, on the basis of the Stanier theory of "simultaneous adaptation", are not quite reliable and may lead to erroneous conclusions.

[Preliminary metabolism of D-ribose by Candida bombi]. / O podgotovitel'nom metabolizme D-ribozy drozhzhami Candida bombi

The activity of D-ribokinase increases and its constitutive synthesis takes place in the yeast Candida bombi after its adaptation to D-ribose. The activity of the enzymes of preparatory metabolism of D-ribose via the reductive pathway after the adaptation to this pentose is low and no growth is found in the medium containing ribitol; therefore, metabolism of D-ribose in C. bombi proceeds via the kinase pathway. D.ribokinase of C. bombi is briefly characterized.