Showing papers in "Applied Biochemistry and Microbiology in 2012"
TL;DR: It is worthwhile to say that algal biofuel production is thought to help stabilize the concentration of carbon dioxide in the atmosphere and decrease global warming impacts.
Abstract: With the decrease of fossil based fuels and the environmental impact of them over the planet, it seems necessary to seek the sustainable sources of clean energy. Biofuels, is becoming a worldwide leader in the development of renewable energy resources. It is worthwhile to say that algal biofuel production is thought to help stabilize the concentration of carbon dioxide in the atmosphere and decrease global warming impacts. Also, among algal fuels’ attractive characteristics, algal biodiesel is non toxic, with no sulfur, highly biodegradable and relatively harmless to the environment if spilled. Algae are capable of producing in excess of 30 times more oil per acre than corn and soybean crops. Currently, algal biofuel production has not been commercialized due to high costs associated with production, harvesting and oil extraction but the technology is progressing. Extensive research was conducted to determine the utilization of microalgae as an energy source and make algae oil production commercially viable.
113 citations
TL;DR: Data is summarized on low-molecular-weight antimicrobial substances, which are primary products or by-products of lactic fermentation, which have long been used for the preparation of cultured dairy products.
Abstract: The review is devoted to literature data on antimicrobial metabolites produced by lactic acid bacteria (LAB), which have long been used for the preparation of cultured dairy products. This paper summarizes data on low-molecular-weight antimicrobial substances, which are primary products or by-products of lactic fermentation. Individual sections are devoted to a variety of antifungal agents and bacteriocins produced by LAB; their potential use as food preservatives has been discussed. The characteristics and classification of bacteriocins are presented in a greater detail; their synthesis and mechanism of action are described using the example of nisin A, which belongs to class I lantibiotics synthesized by the bacterium Lactococcus lactis subsp. lactis. The mechanism of action of class II bacteriocins has been demonstrated with lacticin. Prospective directions for using LAB antimicrobial metabolites in industry and medicine are discussed in the Conclusion.
92 citations
TL;DR: Although the attempts of the production of many secondary metabolites in plant cell culture were unsuccessful so far, new insights in biochemistry and physiology of secondary metabolism, particularly in regulation and compartmentation of secondary metabolite synthesis as well as mechanisms of their transport and storage make these approaches promising.
Abstract: Bioactive substances (BAS) of plant origin are known to play a very important role in modern medicine. Their use, however, is often limited by availability of plant resources and may jeopardize rare species of medicinal plants. Plant cell cultures can serve as a renewable source of valuable secondary metabolites. To the date, however, only few examples of their commercial use are known. The main reasons for such a situation are the insufficient production of secondary metabolites and high cultivation costs. It is possible to increase the performance of plant cell cultures by one or two orders of magnitude using traditional methods, such as selection of highly productive strains, optimization of the medium composition, elicitation, and addition of precursors of secondary metabolite biosynthesis. The progress in molecular biology methods brought about the advent of new means for increasing of the productivity of cell cultures based on the methods of metabolic engineering. Thus, overexpression of genes encoding the enzymes involved in the synthesis of the target product or, by contrast, repression of these genes significantly influences the cell biosynthetic capacity in vitro. Nevertheless, the attempts of the production of many secondary metabolites in plant cell culture were unsuccessful so far, probably due to the peculiarities of the cell culture as an artificial population of plant somatic cells. The use of plant organ culture or transformed roots (hairy root) could turn to be a considerably more efficient solution for this problem. The production of plant-derived secondary metabolites in yeast or bacteria transformed with plant genes is being studied currently. Although the attempts to use metabolic engineering methods were not particularly successful so far, new insights in biochemistry and physiology of secondary metabolism, particularly in regulation and compartmentation of secondary metabolite synthesis as well as mechanisms of their transport and storage make these approaches promising.
73 citations
TL;DR: The PHA degradation was accompanied by a decrease in the polymer molecular weight and an increase in the degree of crystallinity, indicating the preferential destruction of the amorphous phase compared to the crystalline one.
Abstract: Biodegradation of microbial linear polymers of hydroxyalkanoic acids (polyhydroxyalkanoates, PHAs) by soil microbial communities of different structures has been studied during two field seasons in different weather conditions. This process was shown to be influenced by the polymer chemical composition, temperature, humidity, and the microbial soil component. The PHA degradation was accompanied by a decrease in the polymer molecular weight and an increase in the degree of crystallinity, indicating the preferential destruction of the amorphous phase compared to the crystalline one. The quantity of the true PHA destructors developing at the surface of the polymer samples was lower than the quantity of accompanying bacteria. The dominant PHA degrading microorganisms under the test conditions were identified as bacteria of the genera Variovorax, Stenotrophomonas, Acinetobacter, Pseudomonas, Bacillus, and Xanthomonas and as micromycetes from Penicillium, Paecilomyces, Acremonium, Verticillium, and Zygosporium.
64 citations
TL;DR: Data on microorganisms that form methanogenic cellulolytic communities, enzyme complexes of anaerobes essential for cellulose fiber degradation, and feedstock pretreatment are reviewed, as biodegradation is hindered in the presence of lignin.
Abstract: Anaerobic microbial conversion of organic substrates to various biofuels is one of the alternative energy sources attracting the greatest attention of scientists. The advantages of biogas production over other technologies are the ability of methanogenic communities to degrade a broad range of substrates and concomitant benefits: neutralization of organic waste, reduction of greenhouse gas emission, and fertilizer production. Cellulose-containing materials are good substrate, but their full-scale utilization encounters a number of problems, including improvement of the quality and amount of biogas produced and maintenance of the stability and high efficiency of microbial communities. We review data on microorganisms that form methanogenic cellulolytic communities, enzyme complexes of anaerobes essential for cellulose fiber degradation, and feedstock pretreatment, as biodegradation is hindered in the presence of lignin. Methods for improving biogas production by optimization of microbial growth conditions are considered on the examples of biogas formation from various types of plant and paper materials: office paper and cardboard.
64 citations
TL;DR: This review is focused on one of the stages of ontogenesis distinctive by its particular tolerance to the action of unfavorable factors and ability to retain the genomic material for a long period of time, i.e., fungal spores.
Abstract: This review is focused on one of the stages of ontogenesis distinctive by its particular tolerance to the action of unfavorable factors and ability to retain the genomic material for a long period of time, i.e., fungal spores. The major part is devoted to the characterization of the specific stage typical for spores, which is called dormancy. Data are presented characterizing the carbohydrate and lipid composition of spores, with special attention being paid to the role of carbohydrate protectors, in particular, trehalose and mannite, as well as to the role of rafts in the process of sporogenesis. The role of special compounds called autoinhibitors and autostimulators in the process of exit from dormancy is discussed. The final section deals with the role of spore seeding material in biotechnological processes. Data on the correlation between the chemical composition of spores, their ability to remain dormant, and the germination process are considered. Special biotechnological approaches are presented for the first; they allow for the preservation of the germinating ability of spores, intensification of sporogenesis, changes in the ratio of final fermentation products, and an increase in their yield.
64 citations
TL;DR: This review summarizes the potential of microalgae, latest developments in the field and analyzes the “pitfalls” in oversimplification of their promise, including their higher photon conversion efficiency, growth all around the year, even in wastewaters, and production of environment friendly biodegradable biofuels.
Abstract: Microalgae and cyanobacteria are potential food and feed, sources of high-value bioactive molecules and biofuels, and find tremendous applications in bioremediation and agriculture. Although few efforts have been undertaken to index the microalgal germplasm available in terms of lipid content, information on suitability of strains for mass multiplication and advances in development of methods for extraction and generating biofuel are scarce. Our review summarizes the potential of microalgae, latest developments in the field and analyzes the “pitfalls” in oversimplification of their promise in the years to come. Microalgae represent “green gold mines” for generating energy; however, the path to success is long and winding and needs tremendous and concerted efforts from science and industry, besides political will and social acceptance for overcoming the limitations. The major advantages of second generation biofuels based on microalgal systems, include their higher photon conversion efficiency, growth all around the year, even in wastewaters, and production of environment friendly biodegradable biofuels.
53 citations
TL;DR: The quercetin-modified chitosan showed an enhancement of plastic, antioxidant and antimicrobial properties as well as of thermal degradability when applied to Opuntia ficus indica cladodes.
Abstract: Quercetin, rutin, naringin, hesperidin and chrysin were tested as substrates for cloroperoxidase to produce reactive quinones to graft onto chitosan. Quercetin and rutin quinones were successfully chemically attached to low molecular weight chitosan. The quercetin-modified chitosan showed an enhancement of plastic, antioxidant and antimicrobial properties as well as of thermal degradability. Finally, chitosan-quercetin films visibly decreased enzymatic oxidation when applied to Opuntia ficus indica cladodes.
44 citations
TL;DR: It can be predicted that fluoride pollution results in selective pressure that leads to the development of fluoride resistant among bacterial populations, probably through the mechanism which involved high affinity anion binding compounds called ionophores.
Abstract: Microorganisms found in industrial effluents and near the sites of the contamination can be used to indicate pollution and detoxify the contaminated water resources. Emergence of xenobiotic resistant bacteria among them might be potential application in bioremediation. The objective of this study was to isolate and characterize fluoride resistant bacteria from soil and water samples of different regions of India. Five isolates were recovered from different samples which were found to be fluoride resistant. Two of them effectively reduced the fluoride from their media. Through the current study it can be predicted that fluoride pollution results in selective pressure that leads to the development of fluoride resistant among bacterial populations, probably through the mechanism which involved high affinity anion binding compounds called ionophores. Resistant microbes may play a bioremediative role by transforming and concentrating these anions so that they are less available and less dangerous.
40 citations
TL;DR: The fungicidal activity of C. elenkinii can be attributed to the presence of 3-acetyl-2-hydroxy-6-methoxy-4-methyl benzoic acid, the first report of a benzoIC acid derivative having fungicidal action in cyanobacteria.
Abstract: An investigation was directed towards biochemical characterization of cyanobacterium Calothrix elenkinii and analysis of the chemical nature and mode of action of its fungicidal metabolite(s) against oomycete Pythium debaryanum. Biochemical characterization of the culture in terms of carbohydrate utilization revealed the facultative nature of C. elenkinii. Unique antibiotic markers were also found for this strain. 16S rDNA sequencing of the strain revealed 98% similarity with Calothrix sp. PCC7101. The fungicidal activity was tested by disc diffusion assay of different fractions of the culture filtrate. A minimum inhibitory concentration of 10 μl was recorded for ethyl acetate fraction of the 7-weeks old culture filtrates. HPLC, followed by NMR spectral analysis demonstrated the presence of a substituted benzoic acid in the ethyl acetate fraction. Microscopic examination revealed distinct granulation, followed by disintegration of the hyphae of Pythium sp., indicating the presence of an active metabolite in the culture filtrates of Calothrix sp. The fungicidal activity of C. elenkinii can be attributed to the presence of 3-acetyl-2-hydroxy-6-methoxy-4-methyl benzoic acid. This is the first report of a benzoic acid derivative having fungicidal activity in cyanobacteria.
35 citations
TL;DR: The results showed that the degradation efficiency of free cells of B. subtilis YB1 and A. niger YF1 was respectively 87.9 and 98.8% in basic medium III containing 2 mg/l of nicosulfuron after inoculation with 1 ml of culture containing 2.3 × 107 CFU ml−1 and incubation for 5 days at 35°C.
Abstract: The optimal degrading conditions for the nicosulfuron degradation by Bacillus subtilis YB1 and Aspergillus niger YF1, and site of their action on nicosulfuron were studied. The results showed that the degradation efficiency of free cells of B. subtilis YB1 and A. niger YF1 was respectively 87.9 and 98.8% in basic medium III containing 2 mg/l of nicosulfuron after inoculation with 1 ml of culture containing 2.3 × 107 CFU ml−1 and incubation for 5 days at 35°C. Moreover, the degradation rate of nicosulfuron by the mixture of microorganisms was much higher than for every of them taken separately in the same conditions. The mass spectrometric analysis of the products degraded by B. subtilis YB1 revealed that the sulfonylurea bridge in nicosulfuron molecule had been broken. Extracellular (EXF) and endocellular (ENF) fractions obtained from bacterium and fungus were tested for the ability to degrade nicosulfuron. The degradation efficiency of fractions extracted from B. subtilis YB1 was 66.8% by EXF and 15.8% by ENF, but neither EXF nor ENF extracted from A. niger YF1 had the activity of degrading nicosulfuron.
TL;DR: This article reviews the most recent results of studies on the mechanism of induced systemic resistance elicited in plants by non-pathogenic bacteria of the genus Pseudomonas and special focus is focused on the controversial issue of the correlation between the synthesis of pathogenesis-related proteins (PRs) and ISR.
Abstract: This article reviews the most recent results of studies on the mechanism of induced systemic resistance (ISR) elicited in plants by non-pathogenic bacteria of the genus Pseudomonas. Several examples of Pseudomonas strains eliciting resistance against fungal phytopathogens in different species of crop plants are presented. Literature data dealing with bacterial elicitors and the effect of their interaction with plant receptors are quoted. Special focus is focused on the controversial issue of the correlation between the synthesis of pathogenesis-related proteins (PRs) and ISR.
TL;DR: In this review, the microbiological aspects of biobutanol production are reflected and the microbial producers of butanol, obtained by genetic modification of Clostridia and other microorganisms, are characterised.
Abstract: This review is written due to an increased interest in the production of energy carriers and basic substrates of the chemical industry from renewable natural resources. In this review, the microbiological aspects of biobutanol production are reflected and the microbial producers of butanol (both natural, i.e., members of the Clostridium genus, and recombinant), obtained by genetic modification of Clostridia and other microorganisms, are characterised.
TL;DR: Various nitrogen removal technologies using this process, namely, the SHARONANAMMOX CANON and DEAMOX BC-DEAMOX, and their practical use are reviewed, and the current views on the biochemistry of the microbial nitrite reduction by ammonium are presented.
Abstract: This review covers various aspects of the process of anaerobic ammonium oxidation by nitrite with the formation of molecular nitrogen called ANAMMOX (ANaerobic AMMonium Oxidation). Anaerobic ammonium oxidizing bacteria are briefly described, including their phylogenetics, habitat, and morphological and physiological characteristics. The current views on the biochemistry of the microbial nitrite reduction by ammonium are presented. The review is focused on biotechnological wastewater treatment based on the ANAMMOX process. Various nitrogen removal technologies using this process, namely, the SHARONANAMMOX CANON and DEAMOX BC-DEAMOX, and their practical use are reviewed. Various types of reactors and set ups using the ANAMMOX process that are applied to the treatment of wastewater are analyzed. Processing methods for slowly growing ANAMMOX bacterial biomass accumulation aimed at subsequent inoculation in reactors are analyzed. The problems and methods for ANAMMOX bacterial biomass immobilization in reactors and on carriers are described. A description and parameters of laboratory and pilot plants utilizing various high-ammonia wastewater are given. Examples of the currently operating full-scale industrial setups with the ANAMMOX process implementation, including those for the complex biochemical treatment of domestic sewage (BC-DEAMOX) constructed by EKOS (Russia) at the Olympic facilities in the Sochi region, are discussed.
TL;DR: Genetic engineering of biosurfactant producing strains with vgb may be an effective method to increase rhamnolipid production.
Abstract: The potential of Pseudomonas aeruginosa expressing the Vitreoscilla hemoglobin gene (vgb) for rhamnolipid production was studied. P. aeruginosa (NRRL B-771) and its transposon mediated vgb transferred recombinant strain, PaJC, were used in the research. The optimization of rhamnolipid production was carried out in the different conditions of cultivation (agitation rate, the composition of culture medium and temperature) in a time-course manner. The nutrient source, especially the carbon type, had a dramatic effect on rhamnolipid production. The PaJC strain and the wild type cells of P. aeruginosa started producing biosurfactant at the stationary phase and its concentration reached maximum at 24 h (838 mg/l−1) and at 72 h (751 mg l−1) of the incubation respectively. Rhamnolipid production was optimal in batch cultures when the temperature and agitation rate were controlled at 30°C and 100 rpm. It reached 8373 mg l−1 when the PaJC cells were grown in 1.0% glucose supplemented minimal media. Genetic engineering of biosurfactant producing strains with vgb may be an effective method to increase its production.
TL;DR: The ability of the strains-destructors of various aromatic compounds to utilize trinitrotoluene (TNT) up to concentration of 70 mg/l was shown and 4-methyl-3,5-dinitroformamide was discovered for the first time upon the TNT destruction by the bacteria strains of Rh dococcus opacus 1G and Rhdococcus sp.
Abstract: The ability of the strains-destructors of various aromatic compounds to utilize trinitrotoluene (TNT) up to concentration of 70 mg/l was shown. An increase in the TNT concentration from 100 to 150 mg/l did not inhibit its conversion rate by the Kocuria palustris RS32 strain. The Acinetobacter sp. VT11 strain utilized TNT as a sole substrate for growth; 3,5-dinitro-4-methyl anilide acetate and 2,6-dinitro-4-aminotoluene were identified as intermediates of TNT degradation by active strains of Pseudomonas sp. VT-7W and Kocuria rosea RS51. At the same time, 4-methyl-3,5-dinitroformamide was discovered for the first time upon the TNT destruction by the bacteria strains of Rhdococcus opacus 1G and Rhdococcus sp. VT-7. The active bacterial strains achieved an 82-90% destruction of TNT when they were introduced into the soil.
TL;DR: Differences are revealed in the accumulation of proteinases activating protein C and proteinases with thrombinand plasmin-like activities in the growth dynamics of producers.
Abstract: Natural isolates of Aspergillus ochraceus micromycetes from soil and plant remains from various regions have been screened. The isolated strains were characterized by similar cultural and morphological features and an identical nucleotide sequence in the ITS1-5,8S-ITS2 region of rDNA. The ability of the extracellular proteinases of A. ochraceus micromycetes to activate protein C of blood plasma has been established. Differences are revealed in the accumulation of proteinases activating protein C and proteinases with thrombinand plasmin-like activities in the growth dynamics of producers.
TL;DR: Possible ways for combining various biological processes of biohydrogen production are described, with a special focus on the factors that influence the efficiency of coupled systems.
Abstract: Possible ways for combining various biological processes of biohydrogen production are described. Some of these processes are being intensively studied now, whereas others are theoretically feasible, but have not been studied. A special focus is on the factors that influence the efficiency of coupled systems.
TL;DR: Antioxidant ionol and inhibitors of the NADPH oxidase and peroxidase significantly reduce the positive influence of SaA and SuA on the heat resistance of wheat coleoptiles.
Abstract: The influence of salicylic (SaA) and succinic (SuA) acids on the generation of reactive oxygen species (ROS) and the heat resistance of wheat (Triticum aestivum L.) coleoptiles has been studied. The treatment of coleoptiles with 10 μM SaA or SuA results in the accumulation of hydrogen peroxide and enhanced formation of a superoxide anion radical. This effect was partially suppressed by both α-naphthol (the NADPH oxidase inhibitor) and salicylhydroxamic acid (peroxidase inhibitor). SaA and SuA cause an increase in the activity of antioxidant enzymes, such as superoxide dismutase, catalase, and soluble peroxidase, and improve the heat resistance of coleoptiles. Antioxidant ionol and inhibitors of the NADPH oxidase and peroxidase significantly reduce the positive influence of SaA and SuA on the heat resistance of wheat coleoptiles. ROS are considered to be intermediates for heat resistance induction in coleoptiles, treated with SaA and SuA; enhanced ROS generation can be caused by an increased activity of the NADPH oxidase and peroxidase.
TL;DR: A new glucan, namely, piptoporane I, with a molecular mass of 270 kDa was isolated from fruiting bodies of Piptoporus betulinis (Bull.:Fr.) Karst and a polysaccharide with such a structure was isolated for the first time from the fungus genus Piptiporus.
Abstract: A new glucan, namely, piptoporane I, with a molecular mass of 270 kDa was isolated from fruiting bodies of Piptoporus betulinis (Bull.:Fr.) Karst. (Fomitopsidacaeae). Using a combination ofphysicochemical methods, it was established that piptoporane I was a branched glucan with a backbone consisting of alpha-( 1->3)-glucopyranose residues substituted at the C-6 position by single residues of beta-D-glucopyranose by 17.3%. A polysaccharide with such a structure was isolated for the first time from the fungus genus Piptoporus.
TL;DR: Comparison of 3 assays of β-galactosidase activity proved that scanning method for the determination of enzyme activity with using ONPG as substrate is simple, fast and reproducible.
Abstract: Beta-galactosidase, encoded by the lacZ gene in E. coli, can cleave lactose and structurally related compounds to galactose and glucose or structurally related products. Its activity can be measured using an artificial substrate, o-nitrophenyl-beta-D-galactopyranoside (ONPG). Miller firstly described the standard quantitative assay of beta-galactosidase activity in the cells of bacterial cultures by disrupting the cell membrane with the permeabilization solution instead of preparing cell extracts. Therefore, beta-galactosidase became one of the most widely used reporters of gene expression in molecular biology to reflect intracellular gene expression difference. But the Miller assay procedure could not monitor the beta-galactosidase reaction in real time and its results were greatly influenced by some operations in the Miller procedure, such as permeabilization time, reaction time and concentration of the cell suspension. A scanning method based on the Miller method to determine the intracellular beta-galactosidase activity in E. coli Tuner (DE3) expressing -galactosidase in real time was developed and the permeabilization time of cells was optimized for that. The comparison of 3 assays of beta-galactosidase activity (Miller, colorimetric and scanning) was made. The results proved that scanning method for the determination of enzyme activity with using ONPG as substrate is simple, fast and reproducible.
TL;DR: The possible mechanism of Agrobacterium transformation of generative plant cells in planta is discussed and the influence of the plant genotype, bacterial strain, vector construction type, inoculation medium composition, and the conditions of plant treatment with Agrob bacteria are analysed.
Abstract: In this review, methods of Agrobacterium TDNA transfer into plant cells in planta are discussed. The main focus is on the technologies of gene transfer into generative plant cells as a part of Agrobacterium TDNA. The influence of the plant genotype, bacterial strain, vector construction type, inoculation medium composition, and the conditions of plant treatment with Agrobacterium on the efficiency of Agrobacterium transformation in planta is analysed. Based on literature and personal experimental data, the possible mech� anism of Agrobacterium transformation of generative plant cells in planta is discussed.
TL;DR: The addition of the EPS preparation increased the freeze-thaw survival of the more susceptible bacteria 1000-10000 times, and protection was at least partially dependent on the protein component.
Abstract: Microbial communities found on the surface of overwintering plants may be exposed to low temperatures as well as multiple freeze-thaw events. To explore the adaptive mechanisms of these epiphytes, with the objective of identifying products for freeze-protection, enrichment libraries were made from frost-exposed leaves. Of 15 identified bacteria from 60 individual clones, approximately half had ice-association activities, with the great majority showing high freeze-thaw resistance. Isolates with ice nucleation activity and ice recrystallization inhibition activity were recovered. Of the latter, two (Erwinia billingiae J10, and Sphingobacterium kitahiroshimense Y2) showed culture and electron microscopic evidence of motility and/or biofilm production. Mass spectrometric characterization of the E. billingiae extracellular polymeric substance (EPS) identified the major proteins as 35 kDa outer membrane protein A and F, supporting its biofilm character. The addition of the EPS preparation increased the freeze-thaw survival of the more susceptible bacteria 1000-10000 times, and protection was at least partially dependent on the protein component.
TL;DR: The results demonstrates the significance of the studies for the development of a technology for the biotransformation of paper waste into biogas and for the need of selection of microbial communities to improve the efficiency of the process.
Abstract: Several active microbial communities that form biogas via decomposition of cellulose and domestic food waste (DFW) were identified among 24 samples isolated from different natural and anthropogenic sources. The methane yield was 190–260 ml CH4/g from microbial communities grown on cellulose substrates, office paper, and cardboard at 37°C without preprocessing. Under mesophilic conditions, bioconversion of paper waste yields biogas with a methane content from 47 to 63%; however, the rate of biogas production was 1.5–2.0 times lower than under thermophilic conditions. When microbial communities were grown on DFW under thermophilic conditions, the most stable and effective of them produced 230–353 ml CH4/g, and the methane content in biogas was 54–58%. These results demonstrates the significance of our studies for the development of a technology for the biotransformation of paper waste into biogas and for the need of selection of microbial communities to improve the efficiency of the process.
TL;DR: It is found that feather protein can be metabolized for production of animal feed protein concentrates and found that protease activity increased by 2-fold in presence of 10 mM Mn2+ whereas Ba2+ and Hg2+ inhibited it.
Abstract: The SN1 strain of Bacillus megaterium, isolated from soil of Ghazipur poultry waste site (India) produced extracellular caseinolytic and keratinolytic enzymes in basal media at 30°C, 160 rpm in the presence of 10% feather. Feathers were completely degraded after 72 h of incubation. The caesinolytic enzyme was separated from the basal media following ammonium sulphate precipitation and ion exchange chromatography. We report 29.3-fold purification of protease after Q Sepharose chromatography. The molecular weight of this enzyme was estimated to be 30 kDa as shown by SDS-PAGE and zymography studies. Protease activity increased by 2-fold in presence of 10 mM Mn2+ whereas Ba2+ and Hg2+ inhibited it. Ratio of milk clotting activity to caseinolytic activity was found to be 520.8 for the 30–60% ammonium sulphate fraction in presence of Mn2+ ion suggesting potential application in dairy industry. Keratinase was purified to 655.64 fold with specific activity of 544.7 U/mg protein and 12.4% recovery. We adopted the strategy of isolating the keratinolytic and caesinolytic producing microorganism by its selective growing in enriched media and found that feather protein can be metabolized for production of animal feed protein concentrates.
TL;DR: The enzyme immunoassay method showed the presence of sterigmatocystin, emodin, mycophenolic acid, citrinin, alternariol, and diacetoxyscirpenol, which occurred regularly and, in most cases, at a frequency of 55 to 100%.
Abstract: The composition of low-molecular biologically active metabolites typical of microscopic fungi has been studied in blastemas of fruticose lichens of the genera Cladonia, Cetraria, Evernia, Bryoria, and Usnes. The enzyme immunoassay method showed the presence of sterigmatocystin, emodin, mycophenolic acid, citrinin, alternariol, and diacetoxyscirpenol, which occurred regularly and, in most cases, at a frequency of 55 to 100%. The highest levels of accumulation were 0.001–0.003% for emodin, 0.0002% for alternariol and citrinin, 0.0001% for sterigmatocystin and mycophenolic acid, and 0.00005% of the weight of air-dry material for diacetoxyscirpenol. Other metabolites (cyclopiazonic acid, ergot alkaloids, ochratoxin A, PR toxin, deoxynivalenol, zearalenone, and fumonisins) were detected in these lichens less frequently (sometimes only upon the expansion of the territory of sampling), and their content was no more than 0.00005%. The peculiarities of the component composition and the levels of accumulation of fungal metabolites in lichens of different taxonomic affiliation were discussed.
TL;DR: The article studies the nitric oxide (NO) levels in the roots of etiolated seedlings of sowing peas using the DAF-2DA fluorescent probe and fluorescent microscopy and the role of NO in plants under the influence of biotic and abiotic factors is discussed.
Abstract: The article studies the nitric oxide (NO) levels in the roots of etiolated seedlings of sowing peas (Pisum sativum L.) using the DAF-2DA fluorescent probe and fluorescent microscopy. Cross sections of roots of 100–150 µm (the site of a root which is 10–15 mm from the apex) are analyzed. It is shown that the level of NO in the roots after 24 h increased by more than a factor of 2 in the versions with NaNO2 and sodium nitroprusside. At feeding the seedlings with KNO3, a peak in the accumulation of NO in the roots (twofold increase) was observed after 30 min. Fertilizing seedlings with L-arginine (2 mM) increased the intensity of the fluorescence of the root sections by more than a factor of 2. The inoculation of seedlings of rhizobia (Rhizobium leguminosarum bv. viceae) contributed to the reduction of NO on the background of the control (H2O) and sodium nitroprusside and nitrogen compounds. Scavengers of NO (2-phenyl-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide (PTIO), hemoglobin) and inhibitors of nitrate reductase and animal NO synthase (sodium tungstate and aminoguanidine hydrochloride) reduced the level of NO in the roots. The results are discussed in relation to the role of NO in plants under the influence of biotic and abiotic factors.
TL;DR: The simultaneous inactivation of the EDP and OBPPP in the 2Δ [pEL-IDO] strain allowed us to achieve a considerable improvement of the process, namely, a 20% reduction in glucose consumption and a 35% decrease in biomass yield at a constant L-isoleucine/4-HIL conversion rate at a level of 88%.
Abstract: The enzymes of the Entner-Doudoroff pathway (EDP) and/or the oxidative branch of the pentose phosphate pathway (OBPPP) have been inactivated in the E. coli 2Δ (ΔsucAB, ΔaceAK, PL-brnQ) [pEL-IDO] strain, which is used for the biotransformation of L-isoleucine into 4-hydroxyisoleucine (4-HIL) by enzymatic hydroxylation. We established that the inactivation of the EDP in the 2Δ [pEL-IDO] strain does not virtually affect the basic parameters of the process, whereas the blocking of the OBPPP reduces the efficiency of biomass accumulation and L-isoleucine transformation in conversion to glucose. However, the simultaneous inactivation of the enzymes of the EDP and OBPPP in the 2Δ [pEL-IDO] strain allowed us to achieve a considerable improvement of the process, namely, a 20% reduction in glucose consumption and a 35% decrease in biomass yield at a constant L-isoleucine/4-HIL conversion rate at a level of 88%.
TL;DR: Rhodococcal strains displaying high adhesive activity at a low temperature, high salinity, and acidity of the cultivation medium have been selected as a result of the present work and have a considerable potential for use in bioremediation of soil and water contaminated by hydrocarbons.
Abstract: 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.
TL;DR: Extracellular β-D-glucosidase was isolated in a homogeneous state from the Penicillium canescens marine fungus and had transglycosylation activity and hydrolysis of β-glycosidic bonds both in glycosides and in glucose disaccharides.
Abstract: Extracellular beta-D-glucosidase was isolated in a homogeneous state from the Penicillium canescens marine fungus. According to SDS-electrophoresis, the molecular weight of the enzyme was 64 kDa and the maximal activity was observed at pH 5.2 and 70 degrees C. Glucosidase catalyzed the hydrolysis of beta-glycosidic bonds both in glycosides and in glucose disaccharides and had transglycosylation activity. The enzyme can be used for the deglycosylation of natural glycosides and in enzymatic synthesis of new carbon-containing compounds.