Changes in the physicochemical and cultural properties of the bacteria Azospirillum baldaniorum Sp245 under the influence of some synthetic coumarins

M. Kanevsky, Vladislav Menukhov, I. Kosheleva, A. Y. Kostritsky, I. V. Kanevskaya, S. Konnova
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引用次数: 1

Abstract

Due to the small number of studies on the role of coumarins in associative symbiotic relationships, some aspects of the influence of synthetic coumarins on the physicochemical and cultural properties of Azospirillum baldaniorum Sp245 were studied for the first time. To reveal the role of hydroxylation in position 7 of the fused aromatic ring – 1-(2-oxo-2H-chromen-3-yl)butan-1,3-dione, comparative studies of the effect of the original and hydroxylated coumarins on the culture of a model strain of azospirilla were carried out. The survival of bacteria after the addition of coumarins was studied by counting CFU on an agar medium. The biofilm formation activity of the culture was assessed using crystal violet. The change in the surface of bacteria under the action of coumarins was studied by the electrical polarizability of bacterial cells on an ELUS electrooptical analyzer (EloSystemGbR, Germany). The yield and monosaccharide composition of extracellular glycopolymers were studied using gas-liquid chromatography.For the first time, an increase in the yield of EPS of bacteria during growth in the presence of 1-(7-hydroxy-2-oxo-2Hchromen-3-yl)butan-1,3-dione by 1.2 and 1.7 times for concentrations of 50 and 100 μM respectively was observed. It has been established for the first time that the hydroxylated substance has a higher antibacterial activity compared to the unsubstituted one. A decrease in the number of viable cells in planktonic culture and inhibition of biofilm growth were revealed. It has been shown by electro-optical analysis that the presence of coumarins in the cultivation medium in all concentrations studied leads to a change in the electrical polarizability of A. baldaniorum Sp245 cells. The use of electrooptical analysis of cell suspensions using monospecific antibodies obtained against lipopolysaccharides of this strain made it possible to reveal the absence of changes in carbohydrate antigenic determinants on the surface of bacterial cells. This is consistent with the data of the analysis of the composition of extracellular polysaccharides by GLC, during which no differences were found in the qualitative composition and ratio of monosaccharides. An increase in the yield of bacterial EPS during growth in the presence of 1-(7-hydroxy-2-oxo-2H-chromen-3-yl) butan-1,3-dione by 1.2 and 1.7 times for concentrations of 50 and 100 μM was shown. The results obtained allow us to consider the changes that have occurred as features of the adaptation of bacteria to the associative conditions of existence.
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几种合成香豆素对氮螺旋菌Sp245理化特性和培养特性的影响
由于有关香豆素在结合共生关系中的作用的研究较少,本文首次研究了合成香豆素对氮螺旋藻Sp245理化和培养特性的影响。为了揭示融合芳香环-1 -(2-氧- 2h -铬-3-基)丁酮-1,3-二酮7位羟基化的作用,对原始香豆素和羟基化香豆素对氮螺旋藻模型菌株培养的影响进行了比较研究。通过在琼脂培养基上计数CFU,研究了添加香豆素后细菌的存活情况。用结晶紫法测定培养物的生物膜形成活性。在ELUS电光分析仪(EloSystemGbR,德国)上,利用细菌细胞的电极化率研究了香豆素作用下细菌表面的变化。采用气液色谱法对胞外糖共聚物的产率和单糖组成进行了研究。首次观察到1-(7-羟基-2-氧- 2hchromen3 -yl)丁烷-1,3-二酮在50 μM和100 μM浓度下的生长过程中,EPS的产量分别增加了1.2倍和1.7倍。首次证实羟基化物质比未取代的羟基化物质具有更高的抗菌活性。浮游培养中活细胞数量减少,生物膜生长受到抑制。电光分析表明,香豆素在培养培养基中的存在导致了a . baldanorum Sp245细胞电极化率的变化。使用针对该菌株脂多糖获得的单特异性抗体对细胞悬浮液进行光电分析,可以揭示细菌细胞表面碳水化合物抗原决定因素的缺乏变化。这与GLC分析胞外多糖组成的数据一致,单糖的定性组成和比例没有差异。在浓度为50 μM和100 μM的条件下,1-(7-羟基-2-氧- 2h -铬-3-基)丁烷-1,3-二酮的生长过程中,细菌EPS的产量分别增加了1.2倍和1.7倍。获得的结果使我们能够考虑作为细菌适应相关生存条件的特征而发生的变化。
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