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Документ Biotechnological potential of the Acinetobacter genus bacteria(2021) Pirog, Tatiana; Lutsay, Dariya; Muchnyk, FainaUntil recently, there were rare scientific reports on the biotechnological potential of non-pathogenic bacteria of the Acinetobacter genus. Although the first reports about the practically valuable properties of these bacteria date back to the 70s and 80s of the twentieth century and concerned the synthesis of the emulsan bioemulsifier. In the last decade, interest in representatives of the Acinetobacter genus as objects of biotechnology has significantly increased. The review presents current literature data on the synthesis by bacteria of this genus of high-molecular emulsifiers, low-molecular biosurfactants of glyco- and aminolipid nature, enzymes (lipase, agarase, chondroitinase), phytohormones, as well as their ability to solubilize phosphates and decompose various xenobiotics (aliphatic and aromatic hydrocarbons, pesticides, insecticides). Prospects for practical application of Acinetobacter bacteria and the metabolites synthesized by them in environmental technologies, agriculture, various industries and medicine are discussed. The data of own experimental studies on the synthesis and biological activity (antimicrobial, anti-adhesive, ability to destroy biofilms) of biosurfactants synthesized by A. calcoaceticus IMV B-7241 strain and their role in the degradation of oil pollutants, including complex ones with heavy metals, are presented. The ability of A. calcoaceticus IMV B-7241 to the simultaneous synthesis of phytohormones (auxins, cytokinins, gibberellins) and biosurfactants with antimicrobial activity against phytopathogenic bacteria allows us to consider this strain as promising for practical use in crop production to increase crop yields.Документ Effect of cations on the activity of NADP+-dependent glutamate dehydrogenase in Acinetobacter calcoaceticus IMV B-7241, Rhodococcus erythropolis IMV Ac-5017 and Nocardia vaccinii IMV B-7405 grown on industrial waste(2021) Pirog, Tatiana; Paliichuk, Olesia; Lutsay, Dariya; Kliuchka (Nykytyuk), Lilia; Shevchuk, TetianaIntroduction. It is studied the activity of NADP+-dependent glutamate dehydrogenase in the presence of mono- and divalent cations (potential activators of this key enzyme of surface-active aminolipids biosynthesis) in A. calcoaceticus IMV B-7241, R. erythropolis IMV Ac-5017 and N. vaccinii IMV B-7405 during cultivation on waste of biodiesel production and sunflower oil waste. Materials and methods. Cultivation of strains was performed in liquid mineral media using as substrates: refined and waste (after frying potato) sunflower oil, refined glycerol and waste of biodiesel production. NADP+-dependent (EC 1.4.1.4) glutamate dehydrogenase activity in cell-free extracts was analyzed for glutamate formation during oxidation of NADPH at 340 nm. Monovalent (Na+, K+) and divalent (Mg2+, Ca2+, Zn2+) cations in the form of salts of NaCl, KCl, MgSO4 × 7H2O, CaCl2 and ZnSO4 × 7H2O were added to the reaction mixture, as well as into the medium for strains cultivation. Results and discussion. Calcium cations were found to be activators of NADP+-dependent glutamate dehydrogenase activity in R. erythropolis IMV Aс-5017 and N. vaccinii IMV B-7405 grown on refined and waste sunflower oil: in the presence of 1–5 mmol Ca2+ in the mixture, the activity of the enzyme increased 1.3–2 times compared with that without these cations. The increase in the concentration of CaCl2 to 0.2−0.4 g/l in oil-containing medium of strains IMV Ac-5017 and IMV B-7405 cultivation was accompanied by an increase in NADP+-dependent glutamate dehydrogenase activity by 1.3–1.5 times compared with that on basic medium. When additional quantity of CaCl2 (0.1−0.2 g/l) was introduced into the medium with purified glycerol for the cultivation of A. calcoaceticus IMV B-7241, an increase in NADP+-dependent glutamate dehydrogenase activity was observed by almost 2.5−3 times compared with those for strain IMV B-7241 on the basic medium. There was no impact of activating cations magnesium, zinc, potassium and sodium on NADP+-dependent glutamate dehydrogenase activity of all strains grown on oil-containing substrates and glycerol of different degrees of purification. Conclusion. The results demonstrate the possibility to increase activity of key enzymes of the biosynthesis of the desired product: the composition of the medium should be modified by changing the content of enzymes’ activators.Документ Factors affecting antibiofilm properties of microbial surfactants(2021) Pirog, Tatiana; Kliuchka (Nykytyuk), Lilia; Lutsay, Dariya; Stabnikov, VictorAntibiofilm activity of surface-active substances (SAS) synthesized by Rhodococcus erythropolis, Acinetobacter calcoaceticus, or Nocardia vaccinia was studied. The strains were cultivated using carbon ethanol, glycerol, hexadecane, sunflower oil, and carbon-containing wastes from biodiesel production as carbon sources. Escherichia coli, Bacillus subtilis, Staphylococcus aureus, Pseudomonas aeruginosa, and yeasts Candida albicans and Candida utilis were used as testing cultures. The antibiofilm activity of biosurfactants depended on the source and concentration of carbon source and duration of cultivation. The addition of calcium cations to media significantly increased antibiofilm activity. Replacement of pure sources of carbon for wastes provided the opportunity to receive biosurfactants with high antibiofilm properties on inexpensive substrates. Destruction of biofilms was the same in cases when supernatant or purified SAS was used. Вивчено антибіоплівкову активність поверхнево-активних речовин (ПАР), синтезованих Rhodococcus erythropolis, Acinetobacter calcoaceticus або Nocardia vaccinia. Штами культивували з використанням в якості джерел вуглецю вуглецевого етанолу, гліцерину, гексадекану, соняшникової олії та вуглецевмісних відходів виробництва біодизеля. Як тест-культури використовували Escherichia coli, Bacillus subtilis, Staphylococcus aureus, Pseudomonas aeruginosa, дріжджі Candida albicans і Candida utilis. Антибіоплівкова активність біосурфактантів залежала від джерела та концентрації джерела вуглецю та тривалості культивування. Додавання до середовищ катіонів кальцію значно підвищувало активність антибіоплівки. Заміна чистих джерел вуглецю на відходи дала можливість отримувати на недорогих субстратах біоПАР з високими антибіоплівковими властивостями. Деструкція біоплівок була однаковою у випадках використання супернатанту або очищеного ПАР.Документ Microbial surfactants in environmental technologies(2015) Pirog, Tatiana; Konon, Anastasia; Savenko, IngaIn the review the literature and own experimental data on the use of microbial surface active glycolipids (rhamno-, sophoro- and trehalose lipids) and lipopeptides for water and soil purification from oil and other hydrocarbons, removing toxic heavy metals (Cu2+, Cd2+, Ni2+, Pb2+), degradation of complex pollution (oil and other hydrocarbons with heavy metals), and the role of microbial surfactants in phytoremediation processes was shown. The factors that limit the use of microbial surfactants in environmental technologies are discussed. Thus, at certain concentrations biosurfactant can exhibit antimicrobial properties and inhibit microorganisms destructing xenobiotics. Microbial biodegradability of surfactants may also reduce the effectiveness of bioremediation.Документ Intensification of surfactants’ synthesis by Rhodococcus erythropolis IMV Ac-5017, Acinetobacter calcoaceticus IMV B-7241 and Nocardia vaccinii K-8 on fried oil and glycerol containing medium(2013) Pirog, Tatiana; Sofіlkanich (Morozova), Anna; Konon, Anastasia; Shevchuk, Tetiana; Ivanov, SergeyАктуальною проблемою сьогодення є пошук шляхів для переробки відходів. Біотехнологія є одним з найбільш екологічно привабливих методів, що дають змогу вирішити проблему утилізації відходів і одержання цінних мікробних продуктів, наприклад, поверхнево-активних речовин (ПАР). Встановлено, пересмажена (відпрацьована) соняшникова олія, олієвмісні відходи (фузи) і гліцерин можуть бути використані для синтезу ПАР Rhodococcus erythropolis IMВ Ac-5017, Acinetobacter calcoaceticus ІМВ В-7241 і Nocardia vaccinii К-8. Внесення глюкози (0,1%) у середовище з відпрацьованою олією (2% об.) супроводжувалося збільшенням у 4 рази кінцевої концентрації поверхнево-активних речовин (6,8 г/л). У разі одночасного добавлення фумарату і цитрату (0,01-0,2%) у середовище культивування штамів IMВ B-7241 і К-8 спостерігали підвищення кількості позаклітинних ПАР у 2-2,5 рази порівняно з вирощуванням без органічних кислот. Збільшення концентрації ПАР штаму IMВ B-7241 зумовлене одночасним функціонуванням двох анаплеротичних шляхів, а також підвищенням у 3-5 разів активності ферментів біосинтезу поверхнево-активних речовин. Культивування на суміші гліцерину і н-гексадекану (0,5-1,0 об.%) супроводжувалося підвищенням у 1,5-3 рази синтезу ПАР. Препарати ПАР штамів IMВ B-7241 (0,15-0,22 мг/мл) і IMВ Ac-5017 (0.61-2.1 мг/мл) проявляли антимікробну активність щодо Escherichia coli IEM-1 (загибель 67% клітин), а також вегетативних клітин (загибель 45-100%) і спор (загибель 75%) Bacillus subtilis БТ-2. Searching for the ways to process waste has become very topical today. Biotechnology is one of the most environmentally attractive methods, which has the ability to solve the problem of waste utilization and to produce the valuable microbial products, for example biosurfactants. We concluded that fried sunflower oil, oil-containing wastes (soapstock) and glycerol can be used for biosurfactant production by Rhodococcus erythropolis IMV Ac-5017, Acinetobacter calcoaceticus IMV B-7241 and Nocardia vaccinii K-8. Glucose addition (0.1%) into the medium with fried oil (2 vol.%) led to a 4-fold increase of final surfactant concentration (6.8 g/L). The simultaneous addition of fumarate and citrate (0.01–0.2%) into the IMV B-7241 and K-8 strains’ cultivating medium was accompanied by an increase of the exocellular biosurfactant quantity by 2–2.5-fold compared to the cultivation without organic acids. An increase in surfactant concentration of IMV B-7241 strain was the result of the simultaneous functioning of two anaplerotic pathways, also resulting in a 3–5-fold increase in activity of biosynthesis enzymes. Cultivating on a mixture of glycerol and n-hexadecane (0.5–1.0 vol.%) led to a 1.5–3-fold increasing surfactant synthesis. Biosurfactant preparations of IMV B-7241 (0.15–0.22 mg/mL) and IMV Ac-5017 (0.61–2.1 mg/mL) strains were effective against Escherichia coli IEM-1 (67% of cell loss), and vegetative (45–100% of cell loss) and spore (75% of spore loss) cells of Bacillus subtilis BT-2.Документ Intensification of biosurfactant synthesis by Acinetobacter calcoaceticus IMV B-7241 on a hexadecane–glycerol mixture(2012) Pirog, Tatiana; Konon, Anastasia; Shevchuk, Tetiana; Bilets, I.The possibility of enhanced biosurfactant (BS) synthesis by the cultivation of Acinetobacter calcoaceticus IMV B-7241 on a mixture of energetically nonequivalent substrates (hexadecane and glycerol) was shown. Based on theoretical calculations of the energy requirements for biomass production and the synthesis of surface-active trehalose monomycolate from the energy-deficient substrate (glycerol), the concentration of the energy-excessive substrate (hexadecane), which increased the efficiency of the substrate carbon conversion to BS, was determined. The synthesis of extracellular BS on a mixture of hexadecane and glycerol in a molar ratio of 1 : 7 at C/N ratio of 30 increased 2.6–3.5-fold compared to that on single_substrate media. Increased BS synthesis by Acinetobacter calcoaceticus IMV B-7241 grown on a hexadecane–glycerol mixture was accompanied by a 1.3–2.4-fold increase in activities of the enzymes involved in their biosynthesis, as well as by simultaneous functioning of two anaplerotic pathways (the glyoxylate cycle and the phosphoenolpyruvate carboxylase reaction). Досліджено можливість підвищення синтезу поверхнево-активних речовин (ПАР) за умов росту Acinetobacter calcoaceticus ІМВ В-7241 на суміші енергетично нерівноцінних субстратів (гексадекану і гліцерину). На основі теоретичних розрахунків енергетичних потреб синтезу поверхнево-активних трегалозомономіколатів і біомаси на енергетично дефіцитному субстраті (гліцерин) визначена концентрація енергетично надлишкового гексадекану, що дає змогу підвищити ефективність конверсії вуглецю використовуваних субстратів у ПАР. За молярного співвідношення концентрацій гексадекану і гліцерину 1:7 та співвідношенні С/N, що дорівнює 30, кількість синтезованих позаклітинних ПАР збільшувалася у 2.6−3.5 рази порівняно з такою на моносубстратах. Підвищення синтезу ПАР A. calcoaceticus ІМВ В-7241 на суміші гексадекану і гліцерину зумовлене збільшенням в 1.3−2.4 рази активності ферментів їх біосинтезу, а також одночасним функціонуванням двох анаплеротичних шляхів (гліоксилатного циклу і фосфоенолпіруват-карбоксилазної реакції).Документ The influence of culture conditions strain Acinetobacter calcoaceticus K-4 on the synthesis of surfactant(2009) Pirog, Tatiana; Karpenko, Elena; Antoniuk, Svitlana; Shevchuk, TetianaIt has been observed that the Acinetobacter calcoaceticus K-4 strain produces surface-active substances (SAS) while growing either on hydrophilic (ethanol) or on hydrophobic substrates (hexadecane). Maximal SAS synthesis (with a conditional SAS concentration of 3.6; emulsifying activity of culture liquid dissolved in 50 times equal to 96%) was detected with growth on an ethanol-containing medium with the addition of urea, yeast autolysate and microelements, C/N ratio 60:1 and 10% inoculate, cultivated on ethanol-containing medium by the end of the exponential phase of growth. With respect to its chemical nature, extracellular SAS synthesized by A. calcoaceticus K-4 growing on ethanol-containing medium under optimal cultivating conditions form a glycolipid–aminolipid complex.