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  • Ескіз
    Документ
    Effect on phytopathogenic microorganisms of surfactants of microbial origin
    (2021) Pirog, Tatiana; Piatetska, Daria; Yarova, Hanna; Iutynska, Galyna
    Biodegradable non-toxic surfactants of microbial origin are multifunctional preparations, which due to antimicrobial activity are promising for use in crop production to control phytopathogenic microorganisms. Studies on the prospects of using microbial surfactants to control the number of phytopathogenic microorganisms are conducted in three directions: laboratory studies of antimicrobial activity of surfactants in vitro, determination of the effect of surfactants on phytopathogens in vegetative experiments in the process of plants growing in a laboratory or greenhouse, post-harvest treatment of fruits and vegetables with solutions of microbial surfactants to extend their shelf life. The review presents literature data on antimicrobial activity of surfactants against phytopathogenic bacteria and fungi in vitro. Antimicrobial activity of surfactants is evaluated by three main parameters: minimum inhibitory concentration, zones of growth retardation of test cultures on agar media and inhibition of growth of test cultures on agar or liquid media. The vast majority of available publications relate to the antifungal activity of surfactant lipopeptides and rhamnolipids, while data on the effect of these microbial surfactants on phytopathogenic bacteria (representatives of the genera Ralstonia, Xanthomonas, Pseudomonas, Agrobacterium, Pectobacterium) are few. The researchers determined the antimicrobial activity of either total lipopeptides extracted with organic solvents from the culture broth supernatant, or individual lipopeptides (iturin, surfactin, fengycin, etc.) isolated from a complex of surfactants, or culture broth supernatant. Lipopeptides synthesized by members of the genus Bacillus exhibit antimicrobial activity on phytopathogenic fungi of the genera Alternaria, Verticillium, Aspergillus, Aureobasidium, Botrytis, Rhizoctonia, Fusarium, Penicillium, Phytophora, Sclerotinia, Curvularia, Colletotrichum, etc. in sufficiently high concentrations. Thus, the minimum inhibitory concentrations of lipopeptides against phytopathogenic fungi are orders of magnitude higher (in average 0.04–8.0 mg/mL, or 40–8000 μg/mL) than against phytopathogenic bacteria (3–75 μg/mL). However, the antifungal activity of lipopeptidecontaining supernatants is not inferior by the efficiency to the activity of lipopeptides isolated from them, and therefore, to control the number of phytopathogenic fungi in crop production, the use of lipopeptidecontaining supernatants is more appropriate. Rhamnolipids synthesized by bacteria of the genus Pseudomonas are more effective antimicrobial agents comparing to lipopeptides: the minimum inhibitory concentrations of rhamnolipids against phytopathogenic fungi are 4–276 μg/mL, which is an order of magnitude lower than lipopeptides. In contrast to the data on the antifungal activity of rhamnolipids against phytopathogens, there are only a few reports in the literature on the effect of these surfactants on phytopathogenic bacteria, whilst the minimal inhibitory concentrations are quite high (up to 5000 μg/mL). The advantage of rhamnolipids as antimicrobial agents compared to lipopeptides is the high level of synthesis on cheap and available in large quantities industrial waste. Currently in the literature there is little information about the effect of surface-active sophorolipids of microbial origin on phytopathogenic fungi, and all these works are mainly about the antifungal activity of sophorolipids. We note that in contrast to surfactant lipopeptides and rhamnolipids, the effective concentration of most sophorolipids, which provides the highest antimicrobial activity against phytopathogens, is higher and reaches 10,000 μg/mL.
  • Ескіз
    Документ
    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.