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Applied Biochemistry and Microbiology (v.41, #5)
The 40th Anniversary of the Skryabin Institute of Biochemistry and Physiology of Microorganisms, Russian Academy of Sciences
by A.M. Boronin (pp. 417-417).
Organic Acid Production by the Yeast Yarrowia lipolytica: A Review of Prospects by T. V. Finogenova; I. G. Morgunov; S. V. Kamzolova; O. G. Chernyavskaya (pp. 418-425).
The review sums up the results of studies of (1) physiological growth characteristics of the yeast Yarrowia lipolytica cultured in the presence of diverse carbon sources (n-alkanes, glucose, and glycerol) and (2) superhigh synthesis of organic acids, which was performed at the Skryabin Institute of Biochemistry and Physiology of Microorganisms, Russian Academy of Sciences. Microbiological processes of obtaining α-ketoglutaric, pyruvic, isocitric, and citric acids are discussed.
Ethanol Metabolism in the Yeasts Yarrowia and Torulopsis: A Review by A. P. Il'chenko; O. G. Chernyavskaya; T. V. Finogenova (pp. 426-432).
Results of research into ethanol metabolism in yeast organisms with highly pronounced aerobic metabolism are reviewed. The low activity of NAD-dependent alcohol dehydrogenase (EC 1.1.1.1), observed under conditions of aerobic yeast growth on ethanol, demonstrates that alternative enzyme systems—alcohol oxidase (EC 1.1.3.13), microsomal ethanol-oxidizing system (including cytochrome P-450), and catalase (EC 1.11.1.6)—may be involved in the alcohol oxidation. The role of these systems in alcohol oxidation and the conditions favoring their operation in this processes are analyzed. It is concluded that iron ions are important regulators of ethanol metabolism for the microorganisms of this group.
Biotechnological Potential of Aerobic Methylotrophic Bacteria: A Review of Current State and Future Prospects by Yu. A. Trotsenko; N. V. Doronina; V. N. Khmelenina (pp. 433-441).
Major results of the authors' findings on the implementation of the biotechnological potential of aerobic methylobacteria and methanotrophs for obtaining forage proteins, biopolymers (polybutyrate and polysaccharides), enzymes (oxidoreductases), and bioprotectors (ectoine), as well as for degrading toxic C1 and Cn compounds, have been reviewed. Unique features of the structural and functional organization of the metabolism of extremophilic (tolerant) methylotrophs are discussed, with a view for their prospective use in various fields of modern biotechnology, including biocatalysis and nanotechnology.
Microbial, Enzymatic, and Immune Biosensors for Ecological Monitoring and Control of Biotechnological Processes by A. N. Reshetilov (pp. 442-449).
Results of the research performed at the Institute of Biochemistry and Physiology of Microorganisms, Russian Academy of Sciences, on designing immunobiosensors for detection of toxic compounds and microbial cells, enzyme-based biosensors for detection of hydrocarbons and alcohols, and microbial biosensors for aromatic compounds, surfactants, and biological oxygen consumption are reviewed. Parameters of the mediator electrodes involving microbial cells and data on the properties of microbial biofuel cells—devices based on the biosensor principle and representing alternative sources of electric energy—are presented.
Enzymes Involved in Modification of the Steroid Nucleus of Industrial Mycobacterial Strains: Isolation, Functions, and Properties by M. V. Donova; V. M. Nikolaeva; O. V. Egorova (pp. 450-456).
The key enzymes involved in modification of the steroid nucleus of sterol-transforming mycobacteria— 3β-hydroxysteroid oxidase (3-OH-SO, EC 1.13.1.2) and 17β-hydroxysteroid dehydrogenase (17-OH-SDH, EC 1.1.1.51)—were isolated and characterized. It is shown that 3-OH-SO is a multifunctional enzyme catalyzing oxidation of the 3β-OH group, Δ5 → Δ4 isomerization, and 6-hydroxylation. Two forms of intracellular 17-OH-SDH that catalyze redox reactions at C17 were found, and their properties were determined. The presence of an extracellular 17-OH-SDH in Mycobacterium spp. (VKM Ac-1815 D and Et1) was demonstrated for the first time.
Degradation of p-Toluenesulfonate by Immobilized Comamonas testosteroni BS1310 (pBS1010) Cells by A. A. Makarenko; A. Yu. Arinbasarova; T. N. Kuvichkina; S. V. Balashov; A. N. Reshetilov (pp. 457-459).
Parameters of degradation of p-toluenesulfonate (TS) by free and agar-embedded Comamonas testosteroni BS1310 (pBS1010) cells were determined. The maximum rate of TS degradation was 25% lower in immobilized than free cells, equaling 11 nmol min−1 mg−1 cells. Degradation of TS by both free and immobilized cells was associated with molecular oxygen consumption (molar ratio 1 : 2). In a plug-flow reactor, the degradation rate was 10.4 nmol min−1 mg−1 cells. The results can be applied to designing reactors for TS degradation in sewage and developing biosensors.
Effects of Naphthalene Degradative Plasmids on the Physiological Characteristics of Rhizosphere Bacteria of the Genus Pseudomonas by O. V. Volkova; T. O. Anokhina; I. F. Puntus; V. V. Kochetkov; A. E. Filonov; A. M. Boronin (pp. 460-464).
We studied the specific growth rate, duration of the lag phase, stability of plasmids, and activities of the key enzymes involved in naphthalene biodegradation in rhizosphere pseudomonades carrying the structurally similar plasmids pOV17 and pBS216. It was demonstrated that these plasmids determined various levels of catechol-2,3-dioxygenase activities. The structural rearrangements in the plasmid pBS216 could “switch off” the genes of the catechol oxidation meta-pathway. It was shown that certain combinations of degradative plasmids and bacterial hosts, such as Pseudomonas chlororaphis PCL1391(pBS216), P. chlororaphis PCL1391(pOV17), and P. putida 53a(pOV17), were considerably more efficient than natural variants in their growth characteristics and the stability of the biodegradation activity, having a potential for bioremediation of soils polluted with polycyclic aromatic hydrocarbons (PAHs).
Degradation of Aniline by Delftia tsuruhatensis 14S in Batch and Continuous Processes by M. S. Sheludchenko; M. P. Kolomytseva; V. M. Travkin; V. N. Akimov; L. A. Golovleva (pp. 465-468).
A Delftia tsuruhatensis strain capable of consuming aniline as the sole source of carbon, nitrogen, and energy at concentrations of up to 3200 mg/l was isolated from activated sludge of the sewage disposal plants of OAO Volzhskii Orgsintez. The strain grew on catechol and p-hydroxybenzoic acid but did not consume phenol, 2-aminophenol, 3-chloroaniline, 4-chloroaniline, 2,3-dichloroaniline, 2,4-dichloroaniline, 3,4-dichloroaniline, 2-nitroaniline, 2-chlorophenol, or aminobenzoate. Aniline is degraded by cleavage of the catechol aromatic ring at the ortho position. Cells were immobilized on polycaproamide fiber. It was shown that the strain degraded aniline at 1000 mg/l in a continuous process over a long period of time.
Bacterial Strain Characterizing by EDTA Requirement by A. D. Satroutdinov; T. I. Chistyakova; E. G. Dedyukhina; E. N. Kaparullina; V. K. Eroshin (pp. 469-473).
A novel strain of bacteria (LPM-4) characterized by a unique EDTA requirement for cell growth was isolated. Suspensions of washed cells of strain LPM-4 degraded EDTA complexes with Ba2+, Mg2+, Ca2+, and Mn2+ at constant rates ( 0.310 ± 0.486 mmol EDTA/(g h)) and Zn-EDTA at an initial rate of 0.137 ± 0.016 mmol EDTA/(g h). The temperature optima for cell growth and EDTA degradation were determined under pH-auxostat cultivation. As compared with the known EDTA-degrading bacteria, strain LPM-4 exhibited a higher specific growth rate (0.095− 1) and lower mass cell yield (0.219 g cells/g EDTA), which is promising for its practical applications for EDTA removal in wastewater treatment plants.
Heterogeneity of Rhodococcus opacus 1CP as a Response to Stress Induced by Chlorophenols by M. P. Kolomytseva; I. P. Solyanikova; E. L. Golovlev; L. A. Golovleva (pp. 474-479).
Dissociation of Rhodococcus opacus 1CP during cultivation in different media (containing phenol and its monochlorinated derivatives as the sole source of carbon and energy) was studied. Three variants of strain 1CP (S1, S2, and R) differing in the morphology of cells and colonies, lipid composition, and manner of growth on phenol and monochlorophenols were isolated. It was shown that 2- and 4-chlorophenols were most actively degraded by the smooth (S) forms of the culture, and that the rough (R) form predominated when the culture was grown in a rich medium. The S forms differed from the R forms of the strain by an increased content of cardiolipin, fatty acids, and phosphatidylethanolamine.
Effects of DNA-damaging Agents on Aerobic Methylobacteria Capable and Incapable of Utilizing Dichloromethane by Yu. E. Firsova; M. L. Torgonskaya; N. V. Doronina; Yu. A. Trotsenko (pp. 480-485).
Methylobacterium dichloromethanicum DM4, a degrader of dichloromethane (DCM), was more tolerant to the effect of H2O2 and UV irradiation than Methylobacterium extorquens AM1, which does not consume DCM. The addition of CH2Cl2 to methylobacteria with active serine, ribulose monophosphate, and ribulose bisphosphate pathways of C1 metabolism, grown on methanol, resulted in a 1.1- to 2.5-fold increase in the incorporation of [α-32P]dATP into DNA by the Klenow fragment (exo−). Since DCM dehalogenase was not induced in this process, the increase in the total lengths of DNA gaps resulted from the action of DCM rather than S-chloromethylglutathione (intermediate of primary dehalogenation). The degree of DNA damage in the presence of CH2Cl2 was lower in DCM degraders than methylobacteria incapable of degrading this pollutant. This suggests that DCM degraders possess a more efficient mechanism of DNA repair.
Bacillus subtilis and Phenotypically Similar Strains Producing Hexaene Antibiotics by E. B. Kudryashova; N. G. Vinokurova; E. V. Ariskina (pp. 486-489).
We studied metabolites synthesized by Bacillus subtilis strains, including the type strain of B. atrophaeus and phenotypically similar cultures. These metabolites were represented by polyene antibiotics with conjugated double bonds. Hexaenes from the strains under study inhibited the growth of phytopathogenic fungi Fusarium culmorum, F. sporotrichiella, F. oxysporum, Botrytis sorokiniana, Alternaria tenui, and Phytophthora infestans. The degree of growth inhibition depended on the test fungus.
Effect of the Proteolytic Enzymes of Bacillus licheniformis and the Lysoamidase of Lysobacter sp. XL1 on Proteus vulgaris and Proteus mirabilis Cells by L. P. Ryazanova; L. A. Ledova; N. V. Tsurikova; O. A. Stepnaya; A. P. Sinitsyn; I. S. Kulaev (pp. 490-494).
Preparations of culture liquid of three Bacullus licheniformis strains (S, 103, and 60.4) and the enzymatic preparation lysoamidase from culture liquid of Lysobacter sp. strain XL1 actively lysed pre-autoclaved cells of the gram-negative bacteria Proteus vulgaris and P. mirabilis. Living Proteus cells treated with these enzymatic preparations were lysed during their subsequent autoclaving. Inoculation of enzyme-treated Proteus cells, taken either separately or in combination with one another and polymyxin B, into a rich medium led to cell repair and restoration of the culture viability.
Corrosion of Pipe Steel Samples and Conjugated Conversion of Sulfur Compounds by Thionic Bacteria Halothiobacillus neapolitanus DSM 15147 by A. V. Vatsurina; T. Z. Esikova; V. P. Kholodenko; M. B. Vainshtein; V. I. Dubkova (pp. 495-498).
The kinetics of conversion of sulfur compounds by Halothiobacillus neapolitanus DSM 15147 bacteria was studied in the presence of steel samples. It was shown that the presence of steel altered the known pathway of sulfur compound oxidation by thionic bacteria. Production of atomic hydrogen via the interaction between biogenic sulfuric acid and steel enhanced secondary production of intermediates and decreased the content of sulfate produced previously. The process was accompanied by pH elevation and continuation of intense growth of the thionic bacterium culture. Thionic bacteria formed a corrosive medium, which caused metal destruction. The protective properties of anticorrosive coatings 225 LS and 640 mk were tested. It was shown that these coatings protected steel from the destructive effect of biogenic sulfuric acid.
The Fungus Penicillium citrinum, Isolated from Permafrost Sediments, as a Producer of Ergot Alkaloids and New Quinoline Alkaloids Quinocitrinines by A. G. Kozlovsky; V. P. Zhelifonova; T. V. Antipova (pp. 499-502).
Quinocitrinines and ergot alkaloids were synthesized by the strain Penicillium citrinum VKM FW-800 as the culture grews. The major part of these secondary metabolites was secreted into the medium. In the phase of growth deceleration, these metabolites were partly absorbed by the producer cells. Zinc ions stimulated both the primary and secondary metabolic processes. Addition of this microelement into the culture medium stimulated biomass accumulation and the synthesis of clavine alkaloids and quinocitrinines.
Biosynthesis of Naphthoquinone Pigments by Fungi of the Genus Fusarium by A. G. Medentsev; A. Yu. Arinbasarova; V. K. Akimenko (pp. 503-507).
We studied the biosynthesis of colored naphthoquinone metabolites by Fusarium decemcellulare, F. graminearum, and F. bulbigenum fungi. Depending on the conditions of cultivation, F. bulbigenum and F. graminearum synthesized bikaverin and aurofusarin, respectively, whereas F. decemcellulare synthesized soluble extracellular naphthoquinones of the naphthazarin structure (javanicin, anhydrojavanicin, fusarubin, anhydrofusarubin, bostricoidin, and novarubin) or extracellular dimeric naphthoquinone aurofusarin. The biosynthesis of naphthoquinone pigments was shown to be the main response of the fungi to stress, observed under conditions of growth inhibition or arrest.
New Approaches to Increasing the Yield of Laccase from Panus tigrinus by A. M. Chernykh; A. A. Leont'evskii; L. A. Golovleva (pp. 508-511).
We optimized the conditions for laccase production by the lignolytic fungus Panus tigrinus 8/18. 2,4-Dimethylphenol was used as an aromatic inducer. Introduction of 2,4-dimethylphenol and 2 mM CuSO4 into a rich medium was followed by a tenfold increase in the yield of this enzyme. Additional treatment of the medium with perftoran (an oxygen-transporting agent) and immobilization of the fungus on polycaproamide fibers significantly increased the activity of laccase in the medium. Thus, optimum conditions for cultivation of P. tigrinus were found, which allowed an increase in laccase activity in the medium 25-fold as compared to that achieved using any other method described previously.
Use of Carbon Isotope Composition for Characterization of Microbial Activity in Arable Soils by A. M. Zyakun; O. Dilly (pp. 512-520).
The effect of glucose on microbial mineralization of soil organic matter (SOM) was studied in arable soil specimens. The fluxes of carbon dioxide generated during this degradation were deduced from differences in the carbon isotope abundance ratios of glucose δ13C = –11.4 per mil) and SOM δ13C = –27.01 per mil). The priming effect of glucose and respiratory quotient (RQ) were taken as indices of activation of SOM-consuming microbiota. The data on microbial mineralization of organic matter in soil obtained in this study show that the addition of a readily consumable substance (glucose) to soil favors SOM degradation and increases the release of carbon dioxide from soil to atmosphere.
Wine Yeast Races Maintained in the Russian Collection of Microorganisms (VKM) by W. I. Golubev (pp. 521-523).
A list of pure cultures of over 70 races of wine yeasts maintained within the All-Russia collection of microorganisms is published. Publications on the isolation and investigation of these races are indicated.
Mycelial Fungi Maintained in the All-Russia Collection of Microorganisms (VKM) by S. M. Ozerskaya; G. A. Kochkina; N. E. Ivanushkina; K. M. Zaprometova; S. S. Eremina (pp. 524-527).
Data on the possibility of using the diversity of mycelial fungi maintained in the All-Russia Collection of Microorganisms (VKM) at the Institute for Biochemistry and Physiology of Microorganisms (Russian Academy of Sciences) for research and biotechnological purposes (as producers) are presented, together with information on additions to the basic collection.