Перегляд за Автор "Lutsay, Dariya"

Зараз показуємо 1 - 10 з 10

Antimicrobial activity of surfactances of bacteria Nocardia, Rhodococcus and Acinetobacter genera
(2021) Pirog, Tatiana; Kliuchka, Igor; Lutsay, Dariya; Kliuchka (Nykytyuk), Lilia; Skrotska, Oksana
It was found that the minimum inhibitory concentrations against bacteria and yeast of Acinetobacter calcoaceticus IMV B-7241, Rhodococcus erythropolis IMV Ac-5017 and Nocardia vaccinii IMV B-7405 surfactants, synthesized on traditional substrates, were 9–120 μg/ml and were within the limits defined for the surfactants known in the world. It was for the first time established that surfactants synthesized by the study strains on wastes of biodiesel production and fried sunflower oil were characterized by high antimicrobial activity against bacteria and yeast (minimum inhibitory concentrations 0.45–120 and 1.9–142 μg/ml respectively). It was found that the added of both live and inactivated Escherichia coli ІEM-1 and Bacillus subtilis BT-2 cells in R. erythropolis IMV Ac-5017 and N. vaccinii IMV B-7405 medium cultivation was accompanied by synthesis of surfactants, minimum inhibitory concentrations of which were several times lower than those showed for surfactants synthesized without competitive microorganisms. The obtained results indicate the possibility of using the studied surfactants as effective antimicrobial agents.
Antimicrobial activity of surfactants of microbial origin
(2019) Pirog, Tatiana; Lutsay, Dariya; Kliuchka (Nykytyuk), Lilia; Beregova (Pokora), Khrystyna
The recent literature data about the antibacterial and antifungal activity of microbial surfactants (lipopeptides synthesized by representatives of genera Bacillus, Paenibacillus, Pseudomonas, Brevibacillus, rhamnolipids of bacteria Pseudomonas, Burkholderia, Lysinibacillus sp., sophorolipids of yeasts Candida (Starmerella) and Rhodotorula), and our own experiments data concerning antimicrobial activity of surfactants synthesized by Acinetobacter calcoaceticus IMB B-7241, Rhodococcus erythropolis IMB Aс-5017 and Nocardia vaccini IMV B-7405 were presented. The analysis showed that lipopeptides were more effective antimicrobial agents compared to glycolipids. Thus, the minimum inhibitory concentrations (MIC) of lipopeptides, ramnolipids and sophorolipids are on average (μg/ml): 1–32, 50–500, and 10–200, respectively. The MIC of surfactants synthesized by the IMV B-7241, IMV Ac-5017 and IMV B-7405 strains are comparable to those of the known microbial lipopeptides and glycolipids. The advantages of glycolipids as antimicrobial agents compared with lipopeptides were the possibility of their synthesis on industrial waste and the high concentration of synthesized surfactants. The literature data and our own results indicate the need to study the influence of microbes’ cultivation conditions on the antimicrobial activity of the final product. Останні літературні дані про антибактеріальну та протигрибкову активність мікробних поверхнево-активних речовин (ліпопептидів, синтезованих представниками родів Bacillus, Paenibacillus, Pseudomonas, Brevibacillus, рамноліпідів бактерій Pseudomonas, Burkholderia, Lysinibacillus sp., софороліпідів дріжджів Candida (Starmerella) і Rhodotorula), та наведено дані власних експериментів щодо антимікробної активності поверхнево-активних речовин, синтезованих Acinetobacter calcoaceticus IMB B-7241, Rhodococcus erythropolis IMB Aс-5017 та Nocardia vaccini IMV B-7405. Аналіз показав, що ліпопептиди є більш ефективними протимікробними агентами порівняно з гліколіпідами. Так, мінімальні інгібуючі концентрації (МІК) ліпопептидів, рамноліпідів і софороліпідів становлять у середньому (мкг/мл): 1–32, 50–500 і 10–200 відповідно. МІК ПАР, синтезованих штамами ІМВ В-7241, ІМВ Ас-5017 та ІМВ В-7405, можна порівняти з показниками відомих мікробних ліпопептидів і гліколіпідів. Перевагами гліколіпідів як протимікробних засобів порівняно з ліпопептидами є можливість їх синтезу на промислових відходах і висока концентрація синтезованих ПАР. Дані літератури та власні результати свідчать про необхідність вивчення впливу умов культивування мікробів на антимікробну активність кінцевого продукту.
Biotechnological potential of the Acinetobacter genus bacteria
(2021) Pirog, Tatiana; Lutsay, Dariya; Muchnyk, Faina
Until 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.
Calcium and magnesium cations influence on antimicrobial and antiadhesive activity of Acinetobacter сalcoaceticus ІMV B-7241 surfactants
(2016) Pirog, Tatiana; Sidor, Inga; Lutsay, Dariya
The aim of the work was to study the effect of calcium and magnesium cations on NADP+-dependent glutamate dehydrogenase activity (key enzyme of biosynthesis of Acinetobacter calcoaceticus ІMV B-7241 surface-active aminolipids) followed by modification of medium composition and determining antimicrobial and antiadhesive activity of synthesized surfactants. The strain IMV B-7241 was grown in medium with ethanol. NADP+-dependent glutamate dehydrogenase activity of the cell-free extract was analyzed using the formation of glutamate in the oxidation of NADPH. Surfactants were extracted from supernatant of cultural liquid by mixture of chloroform and methanol (2:1). Antimicrobial against bacteria properties of the surfactants were determined by index of the minimal inhibitory concentration. The number of attached cells and the degree of biofilm destruction were analyzed spectrophotometrically. It was established that in the presence of 10 mM Cа2+ and Mg2+ NADP+-dependent glutamate dehydrogenase activity in the cell-free extract increased to 1.5 times in comparison with that without cations. Increasing concentration of magnesium sulfate to 0.2 g/l, or adding CaCl2 (0.1 g/l) into cultivation medium of IMV B-7241 strain was accompanied by rise of NADP+-dependent glutamate dehydrogenase activity in 2.4 and 3.0 times respectively, as well as increasing antimicrobial and antiadhesive activity of synthesized surfactants. Minimal inhibitory concentration of surfactants synthesized in modified media against some bacteria was in 1.3–3.5 times, adhesion on abiotic surfaces treated with such surfactants in an average of 5–17% lower, and the degree of biofilm destruction in 7–13% higher as compared to indicators for the surfactant produced in the base medium. The obtained results indicate the possibility of regulating antimicrobial and anti-adhesive activity of surfactants under producer cultivation.
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, Tetiana
Introduction. 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.
Factors affecting antibiofilm properties of microbial surfactants
(2021) Pirog, Tatiana; Kliuchka (Nykytyuk), Lilia; Lutsay, Dariya; Stabnikov, Victor
Antibiofilm 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.
Microbial surface-active substances as antiadhesive agents
(2016) Pirog, Tatiana; Savenko, Inga; Lutsay, Dariya
The literature data of recent years about capacity of biosurfactants synthesized by bacteria (Pseudomonas, Lactobacillus, Bacillus) and fungi (Candida, Trichosporon, Saccharomyces) not only to avert the adhesion of microorganisms on the different materials, but also to destroy formed biofilms on them were presented. The perspective of biosurfactants to prevent pathogens colonization on biotic and abiotic surfaces, that is known, can be a reason of cause and spread of infectious diseases was discussed. The data of our researches about antiadhesive properties of biosurfactants synthesized by Acinetobacter calcoaceticus IMV B-7241, Nocardia vaccinii IMV B-7405 and Rhodococcus erythropolis IMV Ac-5017 were presented.
Regulation of biological activity of surfactants under cultivation of Acinetobacter calcoaceticus IMB B-7241 on glycerol
(2021) Pirog, Tatiana; Lutsay, Dariya; Yarova, Hanna
Introduction. The aim of this research was to study the biological activity of surfactants synthesized by Acinetobacter calcoaceticus IMV B-7241 in the medium with glycerol of various degrees of purification and high content of calcium cations (activators of NADH+-dependent glutamate dehydrogenase, which is a key enzyme of biosynthesis of surface-active aminolipids responsible for antimicrobial activity of surfactant complex). Materials and methods. Cultivation of A. calcoaceticus was carried out in liquid mineral medium using as substrates refined glycerol and the waste from biodiesel production. The base medium did not contain calcium chloride, the content of CaCl2 in the modified medium was 0.1 and 0.2 g/l. Surfactants were extracted from the supernatant of the culture luquid with a modified mixture of Folch. The number of adhered cells and the degree of biofilm destruction in the presence of surfactants was determined by spectrophotometric method, antimicrobial activity of surfactants - by the minimum inhibitory concentration (MIC). Results and discussion. It was found that the additional introduction of 0.1−0.2 g/l of CaCl2 into cultivation medium with refined glycerol was accompanied by the synthesis of surfactants, the MIC of which against bacteria (Bacillus subtilis BТ-2, Enterobacter cloacae C-8, Staphylococcus aureus BМS-1) and yeast (Candida albicans D-6) were 1.01–21.3 μg/ml and were 1.4–29 times lower compared to the MIC of surfactants obtained in base medium (1.83−58.8 μg/ml). The adhesion of test cultures on abiotic materials treated with such surfactants was 8–13% lower, and the degree of biofilms destruction was 5–19% higher compared to the values, established for surfactants obtained on base medium. The increasіng antimicrobial and anti-adhesive activity of surfactants synthesized on waste of biodiesel production was observed only when CaCl2 was introduced into the medium at a concentration of 0.2 g/l. Surfactants synthesized in the presence of calcium cations in the medium with the waste of biodiesel production proved to be more effective destructors of bacterial biofilms in comparison with those obtained in the base medium only at low concentrations (0.7–5.5 μg/ml). Conclusion. The results demonstrate the possibility of regulating the biological activity of A. calcoaceticus IMV B-7241 surfactants by changing in the composition of medium content of calcium cations - activators of NADH+-dependent glutamate dehydrogenase (key enzyme of surface-active aminolipids biosynthesis). Surfactants synthesized under different cultivation conditions of A. calcoaceticus IMB B-7241 on glycerol are more effective biofilm destructors and antimicrobial and anti-adhesive agents compared to the known lipopeptides and rhamnolipids formed on glycerol.
Synthesis and biological activity of Аcinetobacter calcoaceticus IMV B-7241 surfactants depending on monovalent cations content in cultivation medium
(2021) Pirog, Tatiana; Lutsay, Dariya; Shevchuk, Tetiana; Iutynska, Galyna
Microbial surfactants (biosurfactants) are multifunctional preparations due to a combination of physicochemical (reduction of surface and interfacial tension, emulsifying activity) and biological (antimicrobial and antiadhesive activity, the ability to destroy biofilms) properties. However, the disadvantage of biosurfactants synthesized as a complex of compounds is the possibility of changing the biological activity depending on the conditions of producer cultivation. Aim. To study the effect of potassium and sodium cations on the NADP+-dependent glutamate dehydrogenase activity of cell-free extract of Acinetobacter calcoaceticus ІМV B-7241 with subsequent appropriate modification of the nutrient medium composition and determination of antimicrobial and anti-adhesive activity of surfactant synthesized. Methods. A. calcoaceticus ІМV B-7241 strain was grown in media containing 2% of sunflower oil waste as a carbon source, as well as various concentrations of potassium and sodium chloride (basal – 1.0 g/l NaCl, medium # 1 that did not contains NaCl, medium # 2 in which the concentration of NaCl was 2.0 g/l, medium # 3 in which the concentration of NaCl and KCl was 1.0 g/l each). The surfactants were extracted from the supernatant liquid culture with a modified Folch mixture. Anti-adhesive activity and the degree of biofilms degradation were determined by spectrophotometric method, antimicrobial activity − by the indicator of the minimum inhibitory concentration (MIC). Activity of enzymes of surface-active aminolipids biosynthesis (NADP+-dependent glutamate dehydrogenase) and glycolipids (phosphoenolpyruvate (PEP) carboxylase, PEP-synthetase, PEP-carboxykinase, trehalose phosphate synthase) were analyzed in cell-free extracts obtained after the destruction of cells by ultrasound. Results. It was found that potassium and sodium cations in concentrations of 50 and 100 mM are inhibitors of NADP+-dependent glutamate dehydrogenase, and in lower concentrations (5–20 mM) – activators of this enzyme, as well as PEP-carboxykinase and PEP-synthetase. The increase in the biosurfactant concentration to 6.1−7.7 g/l during cultivation of A. calcoaceticus ІМV B-7241 in medium # 1 and # 3 was due to the predominant synthesis of glycolipids under such conditions, which was evidenced by the increase in 1.8−6.5 times in the activity of PEP-carboxylase, PEP-carboxykinase, PEP-synthetase and trehalose phosphate synthetase compared to the indicators on the basal medium. The concentration of surfactants synthesized in the basal medium was 3.6 g/l, but such surfactants were characterized by the highest antimicrobial and anti-adhesive activity. Their MIC against the test-cultures of studied bacteria (Pseudomonas sp. MI-2, Bacillus subtilis BT-2, Escherichia coli IEM-1, Staphylococcus aureus BMS-1, Enterobacter cloaceae C-8) and fungi (Candida albicans D-6, Rhizopus nigricans P1, Aspergillus niger P-3, Fusarium culmorum T-7) were 0.88−56 μg/ml and were by 2−3 orders of magnitude lower compared to established for surfactants synthesized in modified media # 1–3. In the case of treatment of abiotic materials with surfactant solutions obtained on the basal medium, the adhesion of bacteria and fungi was on average 10–20 % lower than after surface treatment by the surfactant synthesized in modified media. In the presence of 148−296 μg/ml of surfactants obtained in the basal medium, destruction of S. aureus BMS-1 and B. subtilis BT-2 biofilms was 45−66 %, and C. albicans D-6 yeast – 39−44 %. Under the action of similar concentrations of surfactants synthesized in modified media, the destruction of bacterial and yeast biofilms was lower: 6-52 and 20–46 %, respectively. Conclusions. The obtained results are consistent with the data of our previous studies on the possibility of regulating the antimicrobial and antiadhesive activity of surfactants in the process of producer cultivation by changing the content of cations in the medium, which are inhibitors/activators of enzymes responsible for the synthesis of components of the surfactants complex, which have certain biological properties.
The properties of surfactants synthesized by Acinetobacter calcoaceticus ІMV В-7241 on refined and sunflower oil waste
(2018) Pirog, Tatiana; Lutsay, Dariya; Antoniuk, Svitlana; Elperin, Igor
The aim of the work was to compare the antimicrobial and anti-adhesive activity (including the ability to destroy biofilms), as well as the effect on oil degradation of the surfactants synthesized by the culture of Acinetobacter calcoaceticus IMV B-7241 on refined and sunflower waste. The microbial surfactants were extracted from supernatant of cultural liquid by mixture of chloro form and methanol (2:1). The number of attached cells and the degree of biofilm destruction were analyzed spectrophotometrically. Antimicrobial properties of the surfactants were determined by index of the minimal inhibiting concentration. The concentration of oil in water was analyzed by the gravimetric method after extraction with hexane. It was shown that microbial surfactants synthesized in medium with 2% of both refined and fried oil were characterized by high antimicrobial (MIC with respect to bacterial test cultures 0.8–29 μg/ml, Candida albicans D-6 26−58 μg/ml) and anti-adhesive (decreasing number of bacterial and fungal cells of test cultures attached to abiotic surfaces by 35–70%, destruction of biofilms by an average of 40–44%) activity. Increasing concentration of waste oil in the medium to 4% was accompanied by the formation of surfactants with low antimicrobial activity, in the presence of which the degree of oil destruction in water (3–6 g/l) was 80–88% in 20 days, which is 10–16% higher than when using surfactants synthesized in a medium with 2% oil. Obtained data indicate on the need for studies on the effect of cultivation conditions of producer on the properties of synthesized surfactants for the production of final product with stable predetermined properties, depending on the field of practical application. Метою роботи було порівняти антимікробну та антиадгезійну активність (у тому числі здатність руйнувати біоплівки), а також вплив на деградацію олії ПАР, синтезованих культурою Acinetobacter calcoaceticus ІМВ В-7241 на рафінаді та соняшнику. відходи. Мікробні поверхнево-активні речовини екстрагували із супернатанту культуральної рідини сумішшю хлороформи та метанолу (2:1). Кількість прикріплених клітин і ступінь руйнування біоплівки аналізували спектрофотометрично. Антимікробні властивості ПАР визначали за показником мінімальної інгібуючої концентрації. Концентрацію олії у воді аналізували гравіметричним методом після екстракції гексаном. Показано, що мікробні ПАР, синтезовані на середовищі з 2% як рафінованої, так і смаженої олії, характеризуються високою антимікробною дією (МІК щодо бактеріальних тест-культур 0,8–29 мкг/мл, Candida albicans D-6 26–58 мкг/мл). антиадгезивну (зменшення кількості бактеріальних і грибкових клітин тест-культур, прикріплених до абіотичних поверхонь, на 35–70%, руйнування біоплівок у середньому на 40–44%). Підвищення концентрації відпрацьованої нафти в середовищі до 4 % супроводжувалося утворенням поверхнево-активних речовин із низькою антимікробною активністю, у присутності яких ступінь деструкції нафти у воді (3–6 г/л) у 20 р. становив 80–88 %. днів, що на 10–16 % вище, ніж при використанні ПАР, синтезованих у середовищі з 2 % олії. Отримані дані свідчать про необхідність дослідження впливу умов культивування продуцента на властивості синтезованих ПАР для отримання кінцевого продукту зі стабільними заданими властивостями в залежності від галузі практичного застосування.