[The Factor Stabilizing the Bioluminescence of PVA-Immobilized Photobacteria].

Mikrobiologiia Pub Date : 2017-03-01
L E Aleskerova, K A Alenina, E N Efremnko, A D Ismailov
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Abstract

Immobilization of photobacteria in the cryogel of polyvinyl alcohol (PVA) was carried out. Immobilization was found to result in increased intensity and stability of bioluminescence. The elements determining the stability of bioluminescence were investigated. Selection of the strain was found to be of the highest importance. Among immobilized cells, Photobacterium phosphoreum exhibited the most intense and prolonged light emission, while Vibrio harveyi showed the least one. The technological procedures for cryogenic immobilization of photobacteria were determined. The role of the environment of gel formation in the preservation of the bioluminescence activity was determined. In the gels formed in rich medium for submerged cultivation of photobacteria, almost 100% luminescence activity was preserved, while light emission was considerably prolonged. Bioluminescence intensity of the preparations was shown to depend significantly on pH of the incubation medium. The pH shift to acidic values during prolonged incubation of immobilized cells was shown to be one of the factors of bioluminescence quenching. The stress effects of cryogenic immobilization were found to have an insignificant effect on the temperature profile of bioluminescence. Decreased reduction rate of the luciferase flavin substrate was shown to be a possible reason for bioluminescence quenching.

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聚乙烯醇固定化光细菌生物发光稳定性因素研究
研究了光细菌在聚乙烯醇(PVA)低温凝胶中的固定化。固定化可以增加生物发光的强度和稳定性。研究了影响生物发光稳定性的因素。菌株的选择被认为是最重要的。在固定化细胞中,磷光杆菌的发光强度最大,发光时间最长,而哈维弧菌的发光时间最短。确定了光细菌低温固定化的工艺流程。确定了凝胶形成环境在保存生物发光活性中的作用。在富培养基中形成的凝胶中,光细菌的发光活性几乎保持了100%,而发光时间明显延长。该制剂的生物发光强度与培养液的pH值有显著关系。在固定化细胞的长时间孵育过程中,pH值向酸性值的转变被证明是生物发光猝灭的因素之一。低温固定的应力效应对生物发光的温度分布影响不显著。荧光素酶黄素底物的还原速率降低被证明是生物发光猝灭的可能原因。
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