I. B. Ivshina

Removal of polycyclic aromatic hydrocarbons in soil spiked with model mixtures of petroleum hydrocarbons and heterocycles using biosurfactants from Rhodococcus ruber IEGM 231

Removal of polycyclic aromatic hydrocarbons (PAHs) in soil using biosurfactants (BS) produced by Rhodococcus ruber IEGM 231 was studied in soil columns spiked with model mixtures of major petroleum constituents. A crystalline mixture of single PAHs (0.63g/kg), a crystalline mixture of PAHs (0.63g/kg) and polycyclic aromatic sulfur heterocycles (PASHs), and an artificially synthesized non-aqueous phase liquid (NAPL) containing PAHs (3.00g/kg) dissolved in alkanes C10-C19 were used for spiking. Percentage of PAH removal with BS varied from 16 to 69%. Washing activities of BS were 2.5 times greater than those of synthetic surfactant Tween 60 in NAPL-spiked soil and similar to Tween 60 in crystalline-spiked soil. At the same time, amounts of removed PAHs were equal and consisted of 0.3-0.5g/kg dry soil regardless the chemical pattern of a model mixture of petroleum hydrocarbons and heterocycles used for spiking. UV spectra for soil before and after BS treatment were obtained and their applicability for differentiated analysis of PAH and PASH concentration changes in remediated soil was shown. The ratios A254nm/A288nm revealed that BS increased biotreatability of PAH-contaminated soils.

Soil microbiocenosis as an indicator of stability of meadow communities in the environment polluted with heavy metals

The soil microbiota, a key component of natural ecosystems, is considered as a factor determining the stability of meadow communities. The diversity and abundance of the main ecologically significant groups of microorganisms in meadow soils have been studied along a gradient of long-term soil pollution with heavy metals in the Middle Urals. The results provide evidence for stability of the microbial assemblage formed in these soils. It has been found that the functional activity of certain physiological groups of microorganisms (nitrogen-fixing, denitrifying, and cellulolytic bacteria) and the respiratory activity of microbial communities are stimulated under conditions of heavy-metal soil pollution. Probable effects of the observed changes on mineralization of plant remains in meadow communities are discussed.

Bioconversion of β-Sitosterol and Its Esters by Actinobacteria of the Genus Rhodococcus

The ability of pure cultures of Rhodococcus actinobacteria from the Ural Specialized Collection of Alkanotrophic Microorganisms (World Federation for Culture Collections accession number 768; http://www.ecology.psu.ru/iegmcol) to convert β-sitosterol (BSS) and its 3β-acylated derivatives was studied. Rhodococcus strains with pronounced cholesterol oxidase activity, capable of converting BSS to stigmat-4-ene-3-one in the reaction of cooxidation with n-hexadecane, were selected. The dependence of the activity of cholesterol oxidase of rhodococci on the length of the acyl group in BSS esters was studied. Conditions under which Rhodococcus cells convert BSS to 17β-hydroxyandrost-4-ene-3-one (testosterone), commonly used in pharmacology, were determined.

CHEMICAL CONVERSIONS OF BETULONE PRODUCED BY BIOTRANSFORMATION

The possibility of transforming betulone formed by bio-oxidation of betulin by intact R. rhodochrous IEGM 66 cells was studied. The corresponding mono- and diglutaryl derivatives were prepared from betulone and its oxime. Beckmann fragmentation of 28-benzoyloxy-3-hydroxyiminolup-20(29)-ene synthesized 3,4-secolupane derivatives.

Adaptation of coimmobilized Rhodococcus cells to oil hydrocarbons in a column bioreactor

A possible adaptation of the association of Rhodococcus ruber and Rhodococcus opacus strains immobilized on modified sawdust to oil hydrocarbons in a column bioreactor was investigated. In the bioreactor, the bacterial population showed higher hydrocarbon and antibiotic resistance accompanied by the changes in cell surface properties (hydrophobicity, electrokinetic potential) and in the content of cellular lipids and biosurfactants. The possibility of using adapted Rhodococcus strains for the purification of oil-polluted water in the bioreactor was demonstrated.

Effect of cultivation conditions on the adhesive activity of Rhodococcus cells towards n-Hexadecane

The effect of cultivation conditions (the composition, acidity, and salinity of the cultivation medium; temperature; and the hydrodynamic conditions of cultivation) on the adhesion of actinobacteria of the genus Rhodococcus to n-hexadecane has been investigated. A study performed showed that the adhesive activity of rhodococci depends on the composition of the cultivation medium and on the cultivation temperature. The possible mechanisms underlying the effect of growth conditions on the adhesion of rhodococci to liquid hydrocarbons and involving changes in the cell lipid content or the zeta potential of cells are addressed. Rhodococcal strains displaying high adhesive activity (80–90%) at a low temperature (18°C), high salinity (5.0% NaCl), and acidity (pH 6.0) of the cultivation medium have been selected as a result of the present work; these strains have a considerable potential for use in bioremediation of soil and water contaminated by hydrocarbons.

Methods of microorganism immobilization for dynamic atomic-force studies (review)

Atomic-force microscopy (AFM) is an efficient method for studying the surface ultrastructure and nanomechanical properties of biological objects, including microorganisms. A correctly selected method of microorganism immobilization that provides a strong attachment of cells on the surface of a biologically inert substrate and preservation of their native properties is important for AFM scanning in liquid media. Comparative characteristics of methods of microorganism immobilization applied in dynamic AFM studies are discussed in the review. Technologies of mechanical entrapment and chemical binding of cells to a substrate, as well as protein and immunospecific adsorption, are considered.

The effect of organic solvents on the viability and morphofunctional properties of rhodococcus

The effect of organic solvents on the viability and structural-mechanical (size, surface roughness, and elasticity) properties of actinobacteria of the genus Rhodococcus was studied. The dependence of the survival of Rhodococcus on the hydrophobicity of the solvent, the culture density, and the incubation medium composition was demonstrated. The most resistant strains have been selected. Using combined confocal laser scanning and atomic force microscopy, we established that exposure to solvents resulted in an increase in the relative area and roughness of the surface of living cells and a decrease in their Young’s modulus. The involvement of protonand sodium-dependent efflux pumps in the formation of solvent tolerance in Rhodococcus was demonstrated.

Bioconversion of ecotoxic dehydroabietic acid using Rhodococcus actinobacteria

Actinobactrial strains Rhodococcus erythropolis IEGM 267 and R. rhodochrous IEGM 107 were used to study biodegradation of dehydroabietic acid (DHA), a toxic tricyclic diterpenoid. The experiments were carried out in batch cultures of pre-grown rhodococci in the presence of 0.1% (v/v) n-hexadecane under aerobic conditions for 7 days. It was shown that R. erythropolis IEGM 267 and R. rhodochrous IEGM 107 partially and completely degraded DHA (500 mg/L), respectively. Characteristic physicochemical (reduced zeta potential) and morphological-physiological (increased average size of single cells and cell aggregates, increased root-mean-square roughness) changes in DHA-exposed actinobacteria were revealed. Products of DHA bioconversion by R. erythropolis IEGM 267 were analyzed and exhibited a previously unidentified metabolite 5α-hydroxy-abieta-8,11,13-triene-18-oat. The obtained experimental data widen the knowledge on the catalytic activity of rhodococci and their possible contribution to decontamination of natural ecosystems from pollutants.

The Structural and Functional State of Soil Microbiota in a Chemically Polluted Environment

The structural and functional diversity of the main ecological trophic groups of soil microorganisms in meadow soils of the Central Urals anthropogenically contaminated with heavy metals was studied. The increase in the total numbers of these microorganisms in technozems, in comparison with those in agrozems, is due to the higher abundance of iron-reducing, denitrifying, nitrogen-fixing, and sulfate-reducing bacteria, an increase in cellulolytic activity, and the dependence of these characteristics on the toxic load of the soil. A reductive structure of the microbial community with the predominance of r-strategists, which reflects earlier stages of microbiocenoses succession under soil contamination, is formed under soil pollution with heavy metals.