REAGENT GENERATING SUPEROXIDE ANION AND ITS APPLICATION

摘要

A reagent generating superoxide anion is provided to improve production yield and stability of superoxide anion by inhibiting generation of hydrogen peroxide, so that the reagent is useful for screening superoxide anion-removing materials and for analyzing activity of superoxide dismutase. A reagent generating superoxide anion contains BchZ protein generating superoxide anion with generation of hydrogen peroxide, represented by SEQ ID NO:2, and derived from photosynthetic bacteria. A superoxide anion-removing material is screened by contacting a candidate superoxide anion-removing material and luminal derivatives with the superoxide anion-generating reagent, and measuring luminescence intensity. The activity of superoxide dismutase is analyzed by adding cytochrome C or NBT(nitro blue tetrazolium) into the superoxide anion-generating reagent to induce reduction of cytochrome C or NBT, and measuring absorbance changes of the reaction product. Further, a hemin is connected with the BchZ protein.