Purification of cytochrome P450 BM-3 as a monooxygenase.
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.
Jun
Huang
H
Department of Chemical and Biochemical Engineering, Zhejiang University, Hangzhou, PR China.
Lehe
Mei
M
Qing
Sheng
S
Dongqiang
Lin
L
Shanjing
Yao
Y
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
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