The use of13C/12C isotope abundance ratio for characterization of the origin of ethyl alcohol

The carbon isotope composition of ethyl alcohol produced during alcohol fermentation depended on the substrate used and was characterized by the value of δ¹³C equal to −24.7 ± 0.8%. (wheat grain), −22 ± 0.1%. (rye grain), −22 ± 0.5%. (products of wood hydrolysis), −15.3 ±0.3%. (maize grain) and −10 ± 0.1%. (sugar cane). The isotope composition of carbon of ethyl alcohol obtained during catalytic hydroxylation of ethylene has a δ¹³C of −30.6 ± 0.3%.. The possibility of quantitative determination of specific components in mixtures of ethanol samples with various isotope compositions (chemical synthesis and alcohol fermentation of raw material from C₃ or C₄ plants) was shown.     

Use of glucose and carbon isotope fractionation by microbial cells immobilized on solid-phase surface

By the example of glucose uptake by the soil bacteria Pseudomonas aureofaciens BS1393(pBS216) and Rhodococcus sp. 3–30 immobilized on a solid-phase surface (quartz sand), their growth parameters were determined: growth rate (doubling time), total CO₂ production, CO₂ production per cell, lag period with respect to substrate uptake, respiratory quotient. The growth of P. aureofaciens and Rhodococcus sp. on glucose revealed (1) differences of the lag period with respect to substrate (lag time of ～4 h for P. aureofaciens and ～26 h for Rhodococcus sp.); (2) differences between the maximal rates of CO₂ production (～50 μg C-CO₂ g^?1 sand h^?1 for P. aureofaciens and ～8.5 μg C-CO₂ g^?1 sand h^?1 for Rhodococcus sp.); (3) differences in CO₂ production per cell (～1.94 × 10^?9 μM CO₂/CFU for P. aureofaciens and more than ～3.4 × 10^?9 μM CO₂/CFU for Rhodococcus sp.). The kinetics of the metabolic CO₂ isotopic composition was shown to be determined by the difference in the carbon isotopic characteristics of products in the cell. Upon introduction of glucose into the medium (the preparatory stage of the metabolism), the uptake of intracellular ¹³C-depleted products (lipids) is noted; at the stage of the maximal cell growth rate, introduced glucose is mainly metabolized; and at the final stage, upon exhaustion of substrate, the “stored” products—the lipid fraction—get involved in the metabolism. At the maximal rate of glucose uptake, the CO₂ carbon isotopic fractionation coefficient relative to organic products of microbial biosynthesis was determined to be α = 1.009 ± 0.002.     

Microbial Activity and 13C/12C Ratio as Evidence of N-Hexadecane and N-Hexadecanoic Acid Biodegradation in Agricultural and Forest Soils

The dynamics of microbial degradation of exogenous contaminants, n-hexadecane and its primary microbial oxidized metabolite, n-hexadecanoic (palmitic) acid, was studied for topsoils, under agricultural management and beech forest on the basis the changes in O₂ uptake, CO₂ evolution and its associated carbon isotopic signature, the respiratory quotient (RQ) and the priming effect (PE) of substrates. Soil microbial communities in agricultural soil responded to the n-hexadecane addition more rapidly compared to those of forest soil, with lag-periods of about 23 ± 10 and 68 ± 13 hours, respectively. Insignificant difference in the lag-period duration was detected for agricultural (t_lag = 30 ± 13 h) and forest (t_lag = 30 ± 14 h) soils treated with n-hexadecanoic (palmitic) acid. These results demonstrate that the soil microbiota has different metabolic activities for using n-hexadecane as a reductive hydrocarbon and n-hexadecanoic acid as a partly oxidized hydrocarbon. The corresponding δ¹³C of respired CO₂ after the addition of the hydrocarbon contaminants to soils indicates a shift in microbial activity towards the consumption of exogenous substrates with a more complete degradation of n-hexadecane in the agricultural soil, for which some initial contents of hydrocarbons are inherent. It is supposed that the observed deviation of RQ from theoretically calculated value under microbial substrate mineralization is determined by difference in the time (Δt_i) of registration of CO₂ production and O₂ consumption. Positive priming effect (PE) of n-hexadecane and negative PE of n-hexadecanoic (palmitic) acid were detected in agricultural and forest soils. It is suggested that positive PE of n-hexadecane is conditioned by the induction of microbial enzymes that perform hydroxylation/oxygenation of stable SOM compounds mineralized by soil microbiota to CO₂. The microbial metabolism coupled with oxidative decarboxylation of n-hexadecanoic acid is considered as one of the most probable causes of the revealed negative PE value.     

The Use of the [13C]/[12C] Ratio for the Assay of the Microbial Oxidation of Hydrocarbons

The study deals with a comparative analysis of the relative abundances of the carbon isotopes ¹²C and ¹³C in the metabolites and biomass of the Burkholderia sp. BS3702 and Pseudomonas putida BS202-p strains capable of utilizing aliphatic (n-hexadecane) and aromatic (naphthalene) hydrocarbons as sources of carbon and energy. The isotope compositions of the carbon dioxide, biomass, and exometabolites produced during the growth of Burkholderia sp. BS3702 on n-hexadecane (¹³C = –44.6 ± 0.2) were characterized by the values of ¹³C_{CO 2} = –50.2 ± 0.4, ¹³C_biom = –46.6 ± 0.4, and ¹³C_exo = –41.5 ± 0.4, respectively. The isotope compositions of the carbon dioxide, biomass, and exometabolites produced during the growth of the same bacterial strain on naphthalene (¹³C = –21 ± 0.4) were characterized by the isotope effects ¹³C_{CO 2} = –24.1 ± 0.4, ¹³C_biom = –19.2 ± 0.4, and ¹³C_exo = –19.1 ± 0.4, respectively. The possibility of using the isotope composition of metabolic carbon dioxide for the rapid monitoring of the microbial degradation of petroleum hydrocarbons in the environment is discussed.     

Ratio [13C]/[12C] as an index for express estimation of hydrocarbon-oxidizing potential of microbiota in soil polluted with crude oil

The hydrocarbon-oxidizing potential of soil microbiota and hydrocarbon-oxidizing microorganisms introduced into soil was studied based on the quantitative and isotopic characteristics of carbon in products formed in microbial degradation of oil hydrocarbons. Comparison of CO₂ production rates in native soil and that polluted with crude oil showed the intensity of microbial mineralization of soil organic matter (SOM) in the presence of oil hydrocarbons to be higher as compared with non-polluted soil, that is, revealed a priming effect of oil. The amount of carbon of newly synthesized organic products (cell biomass and exometabolites) due to consumed petroleum was shown to significantly exceed that of SOM consumed for production of CO₂. The result of microbial processes in oil-polluted soil was found to be a potent release of carbon dioxide to the atmosphere.     

The Fractionation of Chlorine Isotopes by the Aerobic Methylotrophic Bacterium Methylobacterium dichloromethanicum Grown on Dichloromethane

Methylobacterium dichloromethanicum was found to be able to utilize dichloromethane (DCM) as the source of carbon and energy with the production of biomass, CO₂, and HCl. A comparative analysis of the abundances of the major DCM isotopomers ³⁵Cl₂ ¹²C¹H₂, ³⁵Cl³⁷Cl¹²C¹H₂, and ³⁷Cl₂ ¹²CH₂1H₂ made it possible to estimate the fractionation of chlorine isotopes during the bacterial metabolism of DCM. The kinetic chlorine isotope effects for ³⁵Cl³⁷Cl¹²C¹H₂ (m/z 86) and ³⁷Cl₂ ¹²C¹H₂ (m/z 88) relative to ³⁵Cl₂ ¹²C¹H₂ (m/z 84) were characterized by _86/84 = 1.006 ± 0.002 and _88/84 = 1.023 ± 0.003, respectively. The inference is made that the growth of M. dichloromethanicum on DCM is accompanied by the mass-independent fractionation of the DCM isotopomers.     

Application of <Superscript>13</Superscript>C mineral carbon for assessment of the primary production of organic matter in aquatic environments

The possibility of measuring the rates of light and dark CO₂ assimilation using ¹³C carbonate was demonstrated on Lake Kichier (Marii El). The application of methods utilizing the stable ¹³C and the radioactive ¹⁴C isotopes resulted in comparable values of the rates of light and dark CO₂ fixation. Due to its absolute environmental safety, the method with ¹³C mineral carbon can be recommended as an alternative to radioisotope methods for qualitative measurements of CO₂ fixation rates in aquatic ecosystems.     

Microbiological and isotopic geochemical research in the arid steppe lakes and sor solonchaks of Western Transbaikalia

The rates of microbial processes in the sor solonchaks and the silts of the Western Transbaikalean saline lakes were investigated. The rate of deep CO₂ fixation in the silt of Lake Beloe was as high as 0.19 mg C dm⁻³ day⁻¹. The content of cellulose and protein in the sor solonchaks (for air-dry material) were up to 5.0 and 0.42 mg g⁻¹, respectively. The rates of cellulose decomposition and proteolysis in the silt were 1.08 and 0.96 % day⁻¹, respectively. Sulfate reduction (up to 1.82 mg S dm⁻³ day⁻¹) prevails at the terminal stages of anaerobic decomposition of organic matter in the silt. During the flooding period, methanogenesis producing isotopically light CH₄ probably prevails in the sor solonchaks and arid soils of the region. This suggestion is supported by the surface foaming of the local arid soils after abundant monsoon rains, an increase in the δ¹³C value for soil carbonates to -5.94‰ along the solonchak profile, and a decrease in the ·¹³C value for the soil carbonates formed by methane-oxidizing bacteria to −11.41‰. The seasonal peaks of isotopically heavier (to −16.60‰) organic matter in the bottom sediments of the lake may be explained as the result of activity of heterotrophic sulfate reducers, which exhibited the highest growth rates at the peak sulfate concentrations in the silt.     

Changes of chlorine isotope composition characterize bacterial dehalogenation of dichloromethane

Fractionation of dichloromethane (DCM) molecules with different chlorine isotopes by aerobic methylobacteria Methylobacterium dichloromethanicum DM4 and Albibacter methylovorans DM10; cell-free extract of strain DM4; and transconjugant Iethylobacterium extorquens AI1/pME 8220, expressing the dcmA gene for DCM dehalogenase but unable to grow on DCM, was studied. Kinetic indices of DCM isotopomers for chlorine during bacterial dehalogenation and diffusion were compared. A two-step model is proposed, which suggests diffusional DCM transport to bacterial cells.