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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 tryptophane on synthesis of certain exometabolites by bacteria of genus Acinetobacter, Nocardia, and Rhodococcus and their properties(2022) Pirog, Tatiana; Piatetska, Daria; Zhdanyuk, Valentina; Leonova, Natalia; Shevchuk, TetianaThe efficiency of integrated microbial biotechnologies for obtaining several practically valuable metabolites in one technological process is determined both by the maximum concentration of these substances and their properties. This is especially true for secondary metabolites, the composition and properties of which vary depending on the cultivation conditions of the producer. Aim. To research the eff ect of tryptophan (a precursor of auxin biosynthesis) in the culture media on the synthesis of certain exometabolites by Rhodococcus erythropolis IMV Ac-5017, Acinetobacter calcoaceticus IMV B-7241, and Nocardia vaccinii IMV B-7405 as well as their properties. Methods. R. erythropolis IMV Ac-5017, A. calcoaceticus IMV B-724, and N. vaccinii IMV B-7405 were cultivated in a medium containing refi ned and waste sunflower oil, biodiesel waste, or ethanol as a carbon source. The concentration of tryptophan in the medium was 300 mg/L. Surfactants were extracted from the supernatant of the cultural liquid with a modified Folch mixture. Phytohormones were isolated from the supernatant by sequential extraction with organic solvents after surfactant extraction. Thin-layer chromatography was used for preliminary purification and concentration of phytohormones. Qualitative and quantitative determination of auxins was performed using high-performance liquid chromatography. The antimicrobial activity of surfactants was analysed by the minimum inhibitory concentration. The activity of enzymes of surface-active glycoand aminolipids biosynthesis (phosphoenolpyruvate synthetase, phosphoenolcarboxykinase, and NADP+-dependent glutamate dehydrogenase) was determined spectrophotometrically during the oxidation of NADH or NADP. Results. It was found that the presence of tryptophan in the culture medium of the strains under study did not affect the number of synthesized surfactants, which was 1.80−1.90, 1.55−1.75, and 1.50−1.65 g/L, respectively. At the same time, cultivation of R. erythropolis IMV Ac-5017, A. calcoaceticus IMV B-724, and N. vaccinii IMV B-7405 in the media with tryptophan increased the number of phytohormones: it was higher than the amount of phytohormones synthesized during cultivation without a precursor. The introduction of tryptophan into the culture medium of the strains was accompanied by the formation of surfactants. These compounds showed 2−4 times higher antimicrobial activity against the phytopathogenic bacteria (Agrobacterium tumefaciens UCM B-1000, Pseudomonas syringae UCM B-1027T, Xanthomonas vesicatoria UCM B-1106, Pectobacterium carotovorum UCM B-1075T, Clavibacter michiganensis IMV B-102 and Pseudomonas syringae pv. tomato IMV B-9167) than compounds synthesized on a medium without a precursor. The antimicrobial activity of surfactants synthesized by A. calcoaceticus IMV B-7241 in the presence of tryptophan either did not change compared to that for surfactants obtained without tryptophan, or increased slightly. Data on the activity of surfactant biosynthesis enzymes correlated with the indicators of their antimicrobial activity. In the presence of tryptophan in the culture medium of N. vaccinii IMV B-7405 and R. erythropolis IMV Ac-5017, NADP+-dependent glutamate dehydrogenase activity in the cells of these strains (a key enzyme for biosynthesis of aminolipids responsible for antimicrobial activity) increased almost by 1.4 times compared to that on a tryptophan-free medium. Conclusions. As a result of this work, it was found that the presence of tryptophan in the culture medium of researched strains did not affect the number of surfactants. The antimicrobial activity of surfactants against phytopathogenic bacteria either increased or remained unchanged compared to that established for surfactants synthesized without a precursor of auxin biosynthesis. The obtained data testify to the high efficiency of the potential use of surfactants complex preparations and phytohormones in crop production to stimulate the growth of plants and biocontrol of phytopathogenic bacteria.Документ Synthesis of gibberellins by surfactant producers Nocardia vaccinii IMV В-7405, Acinetobacter calcoaceticus IMV В-7241 and Rhodococcus erythropolis IMV Ас-5017(2020) Leonova, Natalia; Pirog, Tatiana; Piatetska, Daria; Shevchuk, Tetiana; Kharkhota, Maxym; Iutynska, GalynaIn this study, we established that the producers of surfactants Nocardia vaccinii IMV В-7405, Acinetobacter calcoaceticus IMV B-7241 and Rhodococcus erythropolis IMV Аc-5017 were able to synthesize phytohormones of gibberellic nature during cultivation on the traditional substrates (ethanol, hexadecane) and on the industrial waste sunflower oil and biodiesel production waste). The results of specific biotesting showed that the treatment of cucumber seedlings with phytohormonal extracts of all studied strains at 1:500 and 1:600 dilutions stimulated the extension of the cucumber hypocotyls as compared to the water control in values close to the treatment with gibberellic acid. The results of high-performance liquid chromatography showed N. vaccinii IMV B-7405, A. calcoaceticus IMV B-7241 and R. erythropolis IMV Аc-5017 produce highly active forms of gibberellins GA3 and GA4. The level of its synthesis was nearly the same (6.0-10.0 μg∙L-1) under cultivation of strains on every substrate. The exception was strain N. vaccinii IMV В-7405 which synthesized almost 47.0 μg∙L-1 GA3 and GA4 while is growing on the waste oil from meat frying. The obtained results are the groundwork for the development of an economically profitable technology for the recycling of toxic wastes using N. vaccinii IMV B-7405, A.calcoaceticus IMV B-7241 and R. erythropolis IMV Аc-5017. Such technology will allow us to develop complex microbial preparations with various biological properties in a single process. У цьому дослідженні встановлено, що продуценти ПАР Nocardia vaccinii ІМВ В-7405, Acinetobacter calcoaceticus ІМВ В-7241 та Rhodococcus erythropolis ІМВ Ас-5017 здатні синтезувати фітогормони гіберелової природи при культивуванні на традиційних субстратах (етанол, гексадекан) а також на промислових відходах соняшникової олії та відходах виробництва біодизеля). Результати специфічного біотестування показали, що обробка проростків огірка фітогормональними екстрактами всіх досліджуваних штамів у розведенні 1:500 та 1:600 стимулювала подовження гіпокотилів огірка порівняно з контролем у воді в значеннях, близьких до обробки гібереліновою кислотою. . Результати високоефективної рідинної хроматографії показали, що N. vaccinii IMV B-7405, A. calcoaceticus IMV B-7241 і R. erythropolis IMV Аc-5017 продукують високоактивні форми гіберелінів GA3 і GA4. Рівень його синтезу був приблизно однаковим (6,0-10,0 мкг∙л-1) за культивування штамів на кожному субстраті. Виняток становив штам N. vaccinii ІМВ В-7405, який синтезував майже 47,0 мкг∙л-1 ГА3 та ГА4 під час вирощування на відпрацьованому маслі від смаження м’яса. Отримані результати є основою для розробки економічно вигідної технології переробки токсичних відходів з використанням N. vaccinii ІМВ В-7405, A.calcoaceticus ІМВ В-7241 та R. erythropolis ІМВ Ас-5017. Така технологія дозволить розробляти комплексні мікробні препарати з різними біологічними властивостями в одному процесі.Документ Microbial synthesis of phytohormones(2018) Pirog, Tatiana; Iutynska, Galyna; Leonova, Natalia; Beregova (Pokora), Khrystyna; Schevchuk TetyanaThe aim of the review was to analyze current literature data and the results of own studies on the synthesis of auxins, cytokinins, and gibberellins by plant-associated microorganisms (living in rhizosphere, endophytic, nitrogen-fixing, and phytopathogenic), and by those not involved in symbiotic interactions. Many microorganisms can generate phytohormones, and microbial synthesis of indole-3-acetic acid can be enhanced which can be used in producing it instead of extracting it from plants or by chemical synthesis. Recent progress in intensifying the synthesis of gibberellic acid in deep and solid-phase producer cultivation allows substantially reducing the prime cost of biotechnological production of that phytohormone. The ability of microorganisms to simultaneously synthesize phytohormones and other biologically active compounds with antimicrobial, nematocidal, and other various effects enables creating complex polyfunctional microbial preparations with various biological properties for use in crop production to stimulate plant growth and pest control. Метою огляду було проаналізувати сучасні дані літератури і результати власних досліджень синтезу ауксинів, цитокінінів,гіберелінів як асоційованими з рослинами мікроорганізмами (ризосферними, ендофітними, азотфіксувальними, фітопатогенними), так і тими, які не беруть участі у такій взаємодії. Виявлена у широкого кола мікроорганізмів здатність до утворення фітогормонів, а також успіхи у підвищенні ефективності мікробного синтезу індоліл-3-оцтової кислоти свідчать про можливість такого способу її одержання замість екстракції з рослин або хімічного синтезу. Досягнення останнього десятиліття щодо інтенсифікації синтезу гіберелінової кислоти за умов глибинного і твердофазного культивування продуцентів дають змогу суттєво знизити собівартість цього фітогормону, одержуваного біотехнологічним способом. Здатність мікроорганізмів до одночасного синтезу фітогормонів та інших біологічно активних сполук з антимікробною, нематоцидною та ін. активністю підтверджує можливість створення комплексних поліфункціональних мікробних препаратів з різноманітними біологічними властивостями з метою використання у рослинництві для стимуляції росту рослин і контролю чисельності шкідників.