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N. S. Egorov - One of the best experts on this subject based on the ideXlab platform.
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Characteristics and Properties of the Complex of Proteolytic Enzymes of the Thrombolytic Action of the Micromycete Sarocladium strictum
2021Co-Authors: E. I. Kornienko, A. A. Osmolovskiy, N. A. Baranova, V G Kreyer, I B Kotova, N. S. EgorovAbstract:A preparation of thrombolytic enzymes of Micromycete S. strictum 203 was obtained and characterized. The expressed urokinase activity of producer proteinases was determined, and the content of the complex of three alkaline trypsin-like thiol-dependent serine-type proteinases with different isoelectric points (4.5, 7.2 and 11.8) but close molecular weight was detected in the enzyme preparation (about 35 kDa). One of the proteinases (proteinase III) was not glycosylated, and the rest were glycoproteins. The proteinases differed in the spectrum of proteolytic activity in relation to proteins; the thrombus components also turned out to be different. Presumably, the enzymatic urokinase activity causes proteinases to activate plasminogen.
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production of proteinase with plasmin like and prekallikrein activating activity by the Micromycete aspergillus terreus
2018Co-Authors: E S Zvonareva, V. G. Kreier, A. A. Osmolovskiy, N. A. Baranova, I B Kotova, N. S. EgorovAbstract:The effect of nitrogen sources in the fermentation medium and the cultivation conditions on the production of proteinases with plasmin-like and prekallikrein activation activity by Micromycete Aspergillus terreus 2 was investigated. The highest secretion of proteinases was achieved when the Micromycete was cultivated on a medium containing both amine- and mineral nitrogen sources at an initial pH of 5.5 and at 28°С. It was established that the extracellular Micromycete proteinases are equally capable of hydrolyzing fibrin and fibrinogen.
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Production of proteinases with fibrinolytic and fibrinogenolytic activity by a Micromycete Aspergillus ochraceus
2017Co-Authors: A. A. Osmolovskiy, V. G. Kreier, N. A. Baranova, E. D. Rukavitsyna, N. S. EgorovAbstract:Fibrinolytic and fibrinogenolytic activity of surface and submerged cultures of a Micromycete Aspergillus ochraceus L-1 was studied. Extracellular proteinases produced by A. ochraceus L-1 were found to exhibit specificity against fibrin and fibrinogen and no activity of plasminogen activators. The highest activity was observed in the cultures grown at 28°С and initial pH 7.0. Fibrinolytic activity was shown to be somewhat above 25% of the total plasmin-like activity of A. ochraceus L-1 proteinases.
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Properties of extracellular plasmin-like proteases of Aspergillus ochraceus Micromycete
2017Co-Authors: A. A. Osmolovskiy, V. G. Kreier, N. A. Baranova, N. S. EgorovAbstract:The properties of two extracellular proteases of Aspergillus ochraceus VKM F-4104D Micromycete with plasmin-like activity have been studied. It has been shown that the enzymes differ in pI (5.05 and 6.83) and have similar molecular weights (about 32 and 35 kDa), pH optima (pH 9.0–10.00 at 45°C), and specificities of action on a limited set of chromogenic peptide substrates of trypsin-like proteases. According to inhibitory analysis, both enzymes belong to the serine proteases. Their properties appeared to be similar to those of the protease, protein C activator, which is the main proteolytic enzyme of A. ochraceus VKM F-4104D. Most likely, proteases of this micromyсetes are isoenzymes.
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production of proteinase plasminogen activators by Micromycete tolypocladium inflatum k1
2016Co-Authors: T. S. Sharkova, A V Kurakov, E. O. Matveeva, V. G. Kreier, A. A. Osmolovskiy, N. A. Baranova, N. S. EgorovAbstract:The cultivation conditions (cultivation duration, nitrogen sources, and initial pH of the medium) of the strain Tolypocladium inflatum k1, a producer of extracellular proteases, has been optimized to achieve maximal plasminogen activator activity. It has been shown that Tolypocladium inflatum k1 forms at least two proteinases with activator activity towards plasminogen. One of them is thiol-dependent, EDTA-sensitive serine proteinase, which activates plasminogen and does not hydrolyze fibrin, indicating its higher specificity and prospects for the development of new thrombolytic agents.
A. A. Osmolovskiy - One of the best experts on this subject based on the ideXlab platform.
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properties of extracellular proteinase of the Micromycete aspergillus ustus 1 and its high activity during fibrillary proteins hydrolysis
2021Co-Authors: E A Popova, V G Kreyer, S K Komarevtsev, S V Shabunin, A. A. OsmolovskiyAbstract:A new, nonglycosylated, serine proteinase capable of hydrolyzing collagen, elastin, and fibrin was isolated from a complex preparation of proteins formed by the Micromycete A. ustus 1 under solid-state fermentation. The enzyme had a molecular weight of 33 kDa and a pI of 4.6. The pH-optimum (6.0) and temperature optimum (41°C) of the work of this proteinase were also determined.
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Characteristics and Properties of the Complex of Proteolytic Enzymes of the Thrombolytic Action of the Micromycete Sarocladium strictum
2021Co-Authors: E. I. Kornienko, A. A. Osmolovskiy, N. A. Baranova, V G Kreyer, I B Kotova, N. S. EgorovAbstract:A preparation of thrombolytic enzymes of Micromycete S. strictum 203 was obtained and characterized. The expressed urokinase activity of producer proteinases was determined, and the content of the complex of three alkaline trypsin-like thiol-dependent serine-type proteinases with different isoelectric points (4.5, 7.2 and 11.8) but close molecular weight was detected in the enzyme preparation (about 35 kDa). One of the proteinases (proteinase III) was not glycosylated, and the rest were glycoproteins. The proteinases differed in the spectrum of proteolytic activity in relation to proteins; the thrombus components also turned out to be different. Presumably, the enzymatic urokinase activity causes proteinases to activate plasminogen.
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purification of the protease activator of protein c of human blood plasma produced by the Micromycete aspergillus ochraceus vkm f 4104d
2020Co-Authors: S K Komarevtsev, E A Popova, V G Kreyer, Konstantin A Miroshnikov, A. A. OsmolovskiyAbstract:A method has been developed for the purification of a protease activator of protein C in human blood plasma from the culture fluid of Micromycete Aspergillus ochraceus VKM F-4104D. The properties of this protein are similar to those of the protein-C activator from the venom of the snake Agkistrodon contortrix contortrix, which is used in modern laboratory diagnostics of protein C. The method combines fractionation by ammonium sulfate and hydrophobic, ion-exchange, and gel-permeation chromatography. As a result, a highly purified preparation of protein-C activator was obtained with a 350-fold increase in specific activity during the purification process.
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production of proteinase with plasmin like and prekallikrein activating activity by the Micromycete aspergillus terreus
2018Co-Authors: E S Zvonareva, V. G. Kreier, A. A. Osmolovskiy, N. A. Baranova, I B Kotova, N. S. EgorovAbstract:The effect of nitrogen sources in the fermentation medium and the cultivation conditions on the production of proteinases with plasmin-like and prekallikrein activation activity by Micromycete Aspergillus terreus 2 was investigated. The highest secretion of proteinases was achieved when the Micromycete was cultivated on a medium containing both amine- and mineral nitrogen sources at an initial pH of 5.5 and at 28°С. It was established that the extracellular Micromycete proteinases are equally capable of hydrolyzing fibrin and fibrinogen.
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Production of proteinases with fibrinolytic and fibrinogenolytic activity by a Micromycete Aspergillus ochraceus
2017Co-Authors: A. A. Osmolovskiy, V. G. Kreier, N. A. Baranova, E. D. Rukavitsyna, N. S. EgorovAbstract:Fibrinolytic and fibrinogenolytic activity of surface and submerged cultures of a Micromycete Aspergillus ochraceus L-1 was studied. Extracellular proteinases produced by A. ochraceus L-1 were found to exhibit specificity against fibrin and fibrinogen and no activity of plasminogen activators. The highest activity was observed in the cultures grown at 28°С and initial pH 7.0. Fibrinolytic activity was shown to be somewhat above 25% of the total plasmin-like activity of A. ochraceus L-1 proteinases.
N. A. Baranova - One of the best experts on this subject based on the ideXlab platform.
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Characteristics and Properties of the Complex of Proteolytic Enzymes of the Thrombolytic Action of the Micromycete Sarocladium strictum
2021Co-Authors: E. I. Kornienko, A. A. Osmolovskiy, N. A. Baranova, V G Kreyer, I B Kotova, N. S. EgorovAbstract:A preparation of thrombolytic enzymes of Micromycete S. strictum 203 was obtained and characterized. The expressed urokinase activity of producer proteinases was determined, and the content of the complex of three alkaline trypsin-like thiol-dependent serine-type proteinases with different isoelectric points (4.5, 7.2 and 11.8) but close molecular weight was detected in the enzyme preparation (about 35 kDa). One of the proteinases (proteinase III) was not glycosylated, and the rest were glycoproteins. The proteinases differed in the spectrum of proteolytic activity in relation to proteins; the thrombus components also turned out to be different. Presumably, the enzymatic urokinase activity causes proteinases to activate plasminogen.
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production of proteinase with plasmin like and prekallikrein activating activity by the Micromycete aspergillus terreus
2018Co-Authors: E S Zvonareva, V. G. Kreier, A. A. Osmolovskiy, N. A. Baranova, I B Kotova, N. S. EgorovAbstract:The effect of nitrogen sources in the fermentation medium and the cultivation conditions on the production of proteinases with plasmin-like and prekallikrein activation activity by Micromycete Aspergillus terreus 2 was investigated. The highest secretion of proteinases was achieved when the Micromycete was cultivated on a medium containing both amine- and mineral nitrogen sources at an initial pH of 5.5 and at 28°С. It was established that the extracellular Micromycete proteinases are equally capable of hydrolyzing fibrin and fibrinogen.
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Production of proteinases with fibrinolytic and fibrinogenolytic activity by a Micromycete Aspergillus ochraceus
2017Co-Authors: A. A. Osmolovskiy, V. G. Kreier, N. A. Baranova, E. D. Rukavitsyna, N. S. EgorovAbstract:Fibrinolytic and fibrinogenolytic activity of surface and submerged cultures of a Micromycete Aspergillus ochraceus L-1 was studied. Extracellular proteinases produced by A. ochraceus L-1 were found to exhibit specificity against fibrin and fibrinogen and no activity of plasminogen activators. The highest activity was observed in the cultures grown at 28°С and initial pH 7.0. Fibrinolytic activity was shown to be somewhat above 25% of the total plasmin-like activity of A. ochraceus L-1 proteinases.
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Properties of extracellular plasmin-like proteases of Aspergillus ochraceus Micromycete
2017Co-Authors: A. A. Osmolovskiy, V. G. Kreier, N. A. Baranova, N. S. EgorovAbstract:The properties of two extracellular proteases of Aspergillus ochraceus VKM F-4104D Micromycete with plasmin-like activity have been studied. It has been shown that the enzymes differ in pI (5.05 and 6.83) and have similar molecular weights (about 32 and 35 kDa), pH optima (pH 9.0–10.00 at 45°C), and specificities of action on a limited set of chromogenic peptide substrates of trypsin-like proteases. According to inhibitory analysis, both enzymes belong to the serine proteases. Their properties appeared to be similar to those of the protease, protein C activator, which is the main proteolytic enzyme of A. ochraceus VKM F-4104D. Most likely, proteases of this micromyсetes are isoenzymes.
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production of proteinase plasminogen activators by Micromycete tolypocladium inflatum k1
2016Co-Authors: T. S. Sharkova, A V Kurakov, E. O. Matveeva, V. G. Kreier, A. A. Osmolovskiy, N. A. Baranova, N. S. EgorovAbstract:The cultivation conditions (cultivation duration, nitrogen sources, and initial pH of the medium) of the strain Tolypocladium inflatum k1, a producer of extracellular proteases, has been optimized to achieve maximal plasminogen activator activity. It has been shown that Tolypocladium inflatum k1 forms at least two proteinases with activator activity towards plasminogen. One of them is thiol-dependent, EDTA-sensitive serine proteinase, which activates plasminogen and does not hydrolyze fibrin, indicating its higher specificity and prospects for the development of new thrombolytic agents.
Lugauskas Albinas - One of the best experts on this subject based on the ideXlab platform.
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Different drying technologies and alternation of mycobiots in the raw material of Hyssopus Officinalis L
2020Co-Authors: Raila Algirdas, Lugauskas Albinas, Kemzūraitė Aurelija, Zvicevičius Egidijus, Ragažinskienė Ona, Railienė MarijaAbstract:Contamination of medicinal plant mass with mycobiots is one of(he negative factors deteriorating the quality of raw material. In order to evaluate the impact of the Yield processing technologies upon the changes of mycobiots in raw material, the mycobiotic conditions of herb hyssop (Hyssopus officinalis L.) raw material were evaluated under various regimes of active ventilation and optimization of the drying parameters. The impact of ventilation intensity and temperature of drying agent upon the changes and abundance of mycobiota species in medicinal raw material was determined. Irrespective of file temperature of the airflow, the strongest suppressive effect Upon the mycobiotic contamination in Hyssopi herba was produced by the 5,000 airflow. Analysis of the isolated fungi revealed the prevalence of Penicillium, Aspergillus, Alternaria, Cladosporium, Mucor, Rhizopus species in the raw material. In separate samples Botrytis cinerea, Sclerotinia sclerotiorum, Aureobasidium pullulans, Chryosporium merdarium, Cladorrhinum foecundissimum, Ulocladium consortiale, Trichoderma hamatum, T harzianum, Gilmaniella humicola, Talaromyces flavus, Rhizomucor pusillus, Hansfordia ovalispora, Verticicladium trifidum, Trichosporiella cerebriformis Micromycetes were also rather abundant. Detection of the above-mentioned Micromycetes fit herb hyssop samples differed, and partially depended upon the medium used for their isolationChemijos institutasVytauto Didžiojo universitetasŽemės ūkio akademij
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Toxic Micromycetes in grain raw material during its processing
2020Co-Authors: Lugauskas Albinas, Raila Algirdas, Railienė Marija, Raudonienė VitaAbstract:In 2003-2005 Micromycetes were isolated and identified from wheat, barley, rye, buckwheat grain brought into mills or from processing enterprises. Contamination of the produced flour with Micromycete propagules (cfu g(-1)), changes in Micromycete diversity and abundance in the course of flour storage, preparation and baking of bread, production of groats or other food products and fodder were determined. Most attention was given to widely distributed Micromycetes, known producers of toxins: Alternaria alternata, Aspergillus candidus, A. clavatus, A. flavus, A. fumigatus, A. niger, A. oryzae, A. (=Eurotium) repens, Fusarium culmorum, F. equiseti, F. graminearum, F. moniliforme, F. oxvsporum, F. poae, F. sporotrichioides, Penicillium brevicompactum, P. chrysogenum, P. cyclopium, P. daleae, P. expansum, P. funiculosum, P. roqueforti, P. urticae, P. verruculosum, P. viridicatum, Phoma exiqua, Rhizomucor pusillus, Rhizopus stolonifer, Trichothecium roseum. Abilities of these Micromycetes to produce secondary toxic metabolites were determined as well as possible hazard caused to people consuming the contaminated productsVytauto Didžiojo universitetasŽemės ūkio akademij
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Factors determining accumulation of mycotoxin producers in cereal grain during harvesting
2020Co-Authors: Lugauskas Albinas, Raila Algirdas, Zvicevičius Egidijus, Railienė Marija, Novošinskas HenrikasAbstract:During the meteorologically contrasting period of 2003-2005, the contamination of winter wheat, malt barley and fodder barley grain with Micromycetes during grain harvesting and preparation for storage was investigated. Micromycetes of over 70 species ascribed to 16 genera were isolated and identified, the density of their populations in grain was determined. Micromycetes with a population density of >50% were attributed to dominant species. Short biological characteristic, ecological peculiarities of the dominating Micromycetes are provided; factors determining intensity of their development and abilities to synthesise and excrete toxic metabolites are indicated. The importance of grain drying for stabilisation of its contamination with Micromycete propagules is highlighted. It is noted that in grain dried in shaft dryer using air at 90°C the number of cfu (colony forming units) was reduced from 2.2 to 8.2 times. When active ventilation is applied, conditions favourable for the development of Micromycetes remain longest in the upper layers of the mound. The airflow passing through the layer of damp grain inhibits the development of Micromycetes, but an increase of comparative air flow for more than 500 m'-(t-h)"1 did not reduce the abundance of Micromycete cfu. After drying Alternaria alternata, Fusarium avenaceum, F. culmorum, Penidllum verrucosum dominated in wheat grain; Aspergillus flavus, Bipolaris sorokiniana, Fusarium chlamydosporum, F. culmorum, F tricinctum in malts barley grain; Fusarium avenaceum, F. culmorum, F. tricinctum, Alternaria alternata in fodder barley grain. It has been determined that all Micromycetes recorded on grain after drying are potential producers of toxic metabolites, i.e. are hazardous to human healthVytauto Didžiojo universitetasŽemės ūkio akademij
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Application of ozone for reduction of mycological infection in wheat grain
2020Co-Authors: Raila Algirdas, Lugauskas Albinas, Railienė Marija, Steponavičius Dainius, Steponavičenė Aušra, Zvicevičius EgidijusAbstract:In 2004-2005 means were sought to clean grain from microbiological contamination during transportation and storage. For this purpose, grains with a moisture content of 23.2% of the ?Tauras? variety were selected and ventilated daily for 8 hours until grain wetness was reduced to 14.0%. The effect of ventilation duration and ozone impact was evaluated according to the changes in grain contamination with Micromycetes propagules (cfu?g-1), and alternation of Micromycetes species on the grain surface. At drying grains by active ventilation with an ozone ? air mixture, at O3 concentration of 700 ppb, the drying period was reduced by about 20%, and mycological contamination depends on initial grain moisture content (w): when w=15.2%, contamination was reduced by up to 2.2 times, and when w=22.0% ? up to 3 times. At the same time, the composition of Micromycetes species on the grain surface changed significantly: in non-ventilated grain there were detected Micromycetes of 26 species, and in ventilated grain ? of 11 species. Efficient ozone impact was established only when the mound of wet (w > 18.0%) grains was exposed to ozoneVytauto Didžiojo universitetasŽemės ūkio akademij
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Preventive measures reducing superficial mycobiotic contamination of grain
2020Co-Authors: Steponavičius Dainius, Raila Algirdas, Lugauskas Albinas, Steponavičienė Aušra, Kemzūraitė AurelijaAbstract:Search for the preventive measures reducing the accumulation of mycotoxin producers in food raw material was carried out. Active ventilation was used; the impact of the electro-chemically activated air (ozone) and electro-chemically activated water (anolyte) on the Micromycetes prevailing in grain raw material for food (GRMF) was determined. The GRMF was dried by active ventilation using the ozone-air mixture. Ozone (concentration 1250 ppb) disinfects the surface of the raw material and creates conditions unfavourable for the increase of mycobiotic contamination in drying upper layers of the grain mound. Within 8 days the contamination of GRMF in a mound decreased by 50%, while in its lower layers – more than 3 times. Ventilation of the mound with the above-mentioned concentration of the ozone-air mixture has ceased the active functioning of Fusarium avenaceum, F. graminearum, F. poae, F. solani, F. tricinctum F. sporotrichioides Micromycetes and has considerably retarded the development of Alternaria alternata and other fungi. Anolyte (0.05% of chlorine concentration) reduced the mycobiotic contamination of GRMF by almost 2.5 times. The optimal treatment duration is from 0.5 to 1 hour. The optimal technical parameters, allowing the use of these measures for the preparation of grain food safety technologies, were elaborated; they are designed for more efficient protection of human health against Micromycetes and their toxic metabolites, which are abundantly produced and released into the environmentVytauto Didžiojo universitetasŽemės ūkio akademij
V G Kreyer - One of the best experts on this subject based on the ideXlab platform.
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properties of extracellular proteinase of the Micromycete aspergillus ustus 1 and its high activity during fibrillary proteins hydrolysis
2021Co-Authors: E A Popova, V G Kreyer, S K Komarevtsev, S V Shabunin, A. A. OsmolovskiyAbstract:A new, nonglycosylated, serine proteinase capable of hydrolyzing collagen, elastin, and fibrin was isolated from a complex preparation of proteins formed by the Micromycete A. ustus 1 under solid-state fermentation. The enzyme had a molecular weight of 33 kDa and a pI of 4.6. The pH-optimum (6.0) and temperature optimum (41°C) of the work of this proteinase were also determined.
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Characteristics and Properties of the Complex of Proteolytic Enzymes of the Thrombolytic Action of the Micromycete Sarocladium strictum
2021Co-Authors: E. I. Kornienko, A. A. Osmolovskiy, N. A. Baranova, V G Kreyer, I B Kotova, N. S. EgorovAbstract:A preparation of thrombolytic enzymes of Micromycete S. strictum 203 was obtained and characterized. The expressed urokinase activity of producer proteinases was determined, and the content of the complex of three alkaline trypsin-like thiol-dependent serine-type proteinases with different isoelectric points (4.5, 7.2 and 11.8) but close molecular weight was detected in the enzyme preparation (about 35 kDa). One of the proteinases (proteinase III) was not glycosylated, and the rest were glycoproteins. The proteinases differed in the spectrum of proteolytic activity in relation to proteins; the thrombus components also turned out to be different. Presumably, the enzymatic urokinase activity causes proteinases to activate plasminogen.
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purification of the protease activator of protein c of human blood plasma produced by the Micromycete aspergillus ochraceus vkm f 4104d
2020Co-Authors: S K Komarevtsev, E A Popova, V G Kreyer, Konstantin A Miroshnikov, A. A. OsmolovskiyAbstract:A method has been developed for the purification of a protease activator of protein C in human blood plasma from the culture fluid of Micromycete Aspergillus ochraceus VKM F-4104D. The properties of this protein are similar to those of the protein-C activator from the venom of the snake Agkistrodon contortrix contortrix, which is used in modern laboratory diagnostics of protein C. The method combines fractionation by ammonium sulfate and hydrophobic, ion-exchange, and gel-permeation chromatography. As a result, a highly purified preparation of protein-C activator was obtained with a 350-fold increase in specific activity during the purification process.
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fibrinolytic and collagenolytic activity of extracellular proteinases of the strains of Micromycetes aspergillus ochraceus l 1 and aspergillus ustus 1
2016Co-Authors: A. A. Osmolovskiy, N. A. Baranova, V G Kreyer, E A Popova, N. S. EgorovAbstract:It was shown that extracellular proteinases produced by the strains of Micromycetes A. ochraceus L-1 and A. ustus 1 differ by the activity at various pH as well as by the intensity of the effect on fibrillar proteins. It was revealed that the proteinases of A. ochraceus L-1 demonstrated maximum activity during the growth of the producer in the nitrate-free growth medium (the pH of enzyme reaction was 8.0), whereas those of A. ustus 1 showed maximal activity during the growth of the Micromycete in the medium containing sodium nitrate (the pH of enzyme reaction was 6.0). Values of specific fibrinolytic and collagenolytic activities of A. ochraceus L-1 were 2.2 and 1.6 times higher than those of A. ustus 1. At the same time, A. ustus 1 showed very low values of total proteolytic (caseinolytic) activity and had a high ratio of fibrinolytic activity to total proteolytic (caseinolytic) activity (6.92). It makes the strain a promising producer of proteinases, which hydrolyze fibrin and collagen.
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identification of targets for extracellular proteases activating proteins of the haemostatic system produced by Micromycetes aspergillus ochraceus and aspergillus terreus
2015Co-Authors: E S Zvonareva, A. A. Osmolovskiy, N. A. Baranova, V G Kreyer, I B Kotova, N. S. EgorovAbstract:Effects of extracellular proteases of Aspergillus ochraceus and Aspergillus terreus on plasma hemostasis proteins, consist of initiating the activation of prothrombin complex proteins, was detected. Was discovered, that A. ochraceus proteases have a direct influence on protein C and coagulation factor X, and A. terreus proteases causes their activation indirectly through kallikrein system stimulation. The ability of extracellular proteases of Micromycetes activate prekallikrein in human blood plasma on the example of A. terreus was first demonstrated.