Chumakov V N, Osinskaia L F
Vopr Med Khim. 1977 Sep-Oct;23(5):712-6.
The activity of superhydroxide dismutase was estimated by following rate of inhibition of non-enzymatic reduction of tetrazolium salt in reaction with superhydroxide ions, which were continuously generated in a photochemical system. The quantitative estimations were achieved by using para-nitrotetrazolium chloride, which was reduced to formazanes soluble in acetone. Formation of formazanes by the photochemical system (riboflavin + tetramethylethylene diamine + O2 + para-nitrotetrazolium chloride + light) in presence of biological material and cyanide was compared with formation of formazanes in absence of cyanide for specific estimation of Zn, Cu-dependent forms of superhydroxide dismutase. The rate of enzymatic cyanide sensitive inhibition of the photochemical formation of formazanes from paranitrotetrazolium chloride was determined by the activity of Zn, Cu-dependent superhydroxide dismutase in biological material and might be expressed in units of the activity -- un. ac. = 10/(0,026.% inhibition--1,3).
超氧化物歧化酶的活性是通过追踪在光化学系统中持续产生的超氧离子与四唑盐非酶促还原反应的抑制速率来估算的。定量估算是通过使用对硝基四唑氯化物来实现的,它被还原形成可溶于丙酮的甲臜。在生物材料和氰化物存在的情况下,光化学系统(核黄素+四甲基乙二胺+氧气+对硝基四唑氯化物+光)中甲臜的形成与不存在氰化物时甲臜的形成进行比较,以特异性估算锌、铜依赖性超氧化物歧化酶的形式。由对硝基四唑氯化物光化学形成甲臜的酶促氰化物敏感抑制速率由生物材料中锌、铜依赖性超氧化物歧化酶的活性决定,并且可以用活性单位表示——非活性=10/(0.026×抑制率%-1.3)。