title: Purification of cytochrome P450 BM-3 as a monooxygenase.
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.
authors.affiliation: Department of Chemical and Biochemical Engineering, Zhejiang University, Hangzhou, PR China.
authors.forename: Huang
authors.initials: H
authors.lastname: Jun
authors.forename: Mei
authors.initials: M
authors.lastname: Lehe
authors.forename: Sheng
authors.initials: S
authors.lastname: Qing
authors.forename: Lin
authors.initials: L
authors.lastname: Dongqiang
authors.forename: Yao
authors.initials: Y
authors.lastname: Shanjing
mesh_terms: D001426:Bacterial Proteins / Q000302:isolation & purification* / Q000378:metabolism*; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D003577:Cytochrome P-450 Enzyme System / Q000302:isolation & purification* / Q000378:metabolism*; D004926:Escherichia coli / Q000235:genetics; D007475:Ion Exchange Resins; D006899:Mixed Function Oxygenases / Q000302:isolation & purification* / Q000378:metabolism*; D009251:NADPH-Ferrihemoprotein Reductase; D011994:Recombinant Proteins / Q000302:isolation & purification
chemical_list: D001426:Bacterial Proteins; D007475:Ion Exchange Resins; D011994:Recombinant Proteins; D003577:Cytochrome P-450 Enzyme System; D006899:Mixed Function Oxygenases; D009251:NADPH-Ferrihemoprotein Reductase; C052423:flavocytochrome P450 BM3 monoxygenases