Chloroperoxidase(CPO)from Caldariomyces fumago is a highly versatile oxidase.But its application is limited,because of its low accumulated concentration in the biological synthesis,poor dynamic stability,low recovery of high-purity enzyme in purification process and high price.A novel very efficient method has been developed.At first,CPO was precipitated with hydrophilic polymer(PEG 6000)in the high-saturated solution of aluminum sulfate.And then it was extracted by an aqueous two-phase system of PEG/ammonium sulfate.At last,the extracted CPO was purified by Sephadex G-75 chromatographic system.The results showed that:60% saturated(NH4)2SO4 solution could make PEG 6000 precipitate 95% chloroperoxidase.In the aqueous two-phase system CPO was prior to dissolve in bottom phase,its partition coefficient k was lower than 0.228,its yield about 65.2%,its purification was 7.4 times.Finally,the purity of CPO was slightly higher after chromatographic fractionation.The method was also provide a new method for extracting efficiently other trace biological molecules.

The hydrophilic polymer PEG6000 was used to co-precipitate with chloroperoxidase(CPO)in the saturated ammonium sulfate solution.Then,a aqueous two-phase system of PEG/phosphate was formed based on the co-precipitation resolution to extracted higher composition CPO,and the purity CPO sample from Sephadex G-100 chromatographic system was obtained.Results showed that the CPO yield was 85.5% by co-precipitate with PEG in the presence of the phosphate solution.In the aqueous two-phase system,CPO was dissolved in the bottom phase,the partition coefficient k was lower than 0.341,the yield reached 69.1%,and the purification ratio increased 21.57 times.The purity of CPO was further improved to 24.79 times by column chromatography,the total yield of CPO was 37.75%.