Institute of Biophysics SB RAS, Krasnoyarsk, Russia.
Siberian Federal University, Krasnoyarsk, Russia.
Photochem Photobiol. 2017 Mar;93(2):536-540. doi: 10.1111/php.12639. Epub 2016 Oct 17.
A bioluminescent assay based on a system of coupled enzymatic reactions catalyzed by bacterial luciferase and NADH:FMN-oxidoreductase was developed to monitor toxicity and antioxidant activity of bioactive compounds. The assay enables studying toxic effects at the level of biomolecules and physicochemical processes, as well as determining the toxicity of general and oxidative types. Toxic and detoxifying effects of bioactive compounds were studied. Fullerenols, perspective pharmaceutical agents, nanosized particles, water-soluble polyhydroxylated fullerene-60 derivatives were chosen as bioactive compounds. Two homologous fullerenols with different number and type of substituents, C O (OH) and Fe C (OH) O (x + y = 40-42), were used. They suppressed bioluminescent intensity at concentrations >0.01 g L and >0.001 g L for C O (OH) and Fe C (OH) O , respectively; hence, a lower toxicity of C O (OH) was demonstrated. Antioxidant activity of fullerenols was studied in model solutions of organic and inorganic oxidizers; changes in toxicities of general and oxidative type were determined; detoxification coefficients were calculated. Fullerenol C O (OH) revealed higher antioxidant ability at concentrations 10 -10 g L . The difference in the toxicity and antioxidant activity of fullerenols was explained through their electron donor/acceptor properties and different catalytic activity. Principles of bioluminescent enzyme assay application for evaluating the toxic effect and antioxidant activity of bioactive compounds were summarized and the procedure steps were described.
基于细菌荧光素酶和 NADH：FMN-氧化还原酶偶联酶促反应系统，开发了一种生物发光测定法，用于监测生物活性化合物的毒性和抗氧化活性。该测定法能够在生物分子和物理化学过程水平上研究毒性作用，并确定一般和氧化型毒性。研究了生物活性化合物的毒性和解毒作用。富勒醇、有前途的药物制剂、纳米级颗粒、水溶性多羟基富勒烯-60 衍生物被选为生物活性化合物。使用了两种具有不同取代基数量和类型的同系富勒醇，CO(OH)和 FeCO(OH)O(x + y = 40-42)。它们在浓度>0.01 g/L 和>0.001 g/L 时抑制生物发光强度；因此，证明了 CO(OH)的毒性较低。在有机和无机氧化剂的模型溶液中研究了富勒醇的抗氧化活性；确定了一般和氧化型毒性的变化；计算了解毒系数。富勒醇 CO(OH)在 10-10 g/L 的浓度下表现出更高的抗氧化能力。通过它们的电子供体/受体性质和不同的催化活性解释了富勒醇的毒性和抗氧化活性的差异。总结了生物发光酶测定法在评估生物活性化合物毒性作用和抗氧化活性中的应用原则，并描述了步骤程序。
