Purification of cytochrome P450 BM-3 as a monooxygenase |
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Authors: | Jun Huang Lehe Mei Qing Sheng Dongqiang Lin Shanjing Yao |
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Affiliation: | Department of Chemical and Biochemical Engineering, Zhejiang University, Hangzhou, PR China. |
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Abstract: | After investigating two anion-exchange resins, the purification factor and activity yields of P450 BM-3 were higher with Resource Q than with DEAE-Sepharose FF. Screening of HIC media showed that Source 15ISO was the most suitable for purification of P450 BM-3. An effective isolation and purification procedure of P450 BM-3 was developed and included three steps: 35%-70% saturation (NH(4))(2)SO(4) precipitation, Source 15ISO hydrophobic interaction chromatograph and Sephacryl S-200 gel filtration chromatography. Using this protocol, the purification factor and P450 BM-3 activity recovery was 13.5 and 13.7%, respectively. |
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