Cytochrome P450 BM3
Shaun D. Black
shaun at uthct.edu
Thu Jul 23 18:30:07 EST 1998
studer at okstate.edu wrote...
> Does anyone out there use P450 BM3 ? I have a problem removing
>the substrate(mystiric acid). The soret is high spin 420 low spin 392 and
>the indication is that even after centrifuge filtration for 5 volumes and
>25 hours of dialysis the protein and subtrate are still complexed. Idid
>not use size exclusion because i read it removed a protein part and
>consiquently the protein lost activity.
You might run it through our affinity purification once; the procedure
works well to remove all kinds of impurities. Check out Black, et al.,
Arch. Biochem. Biophys. 310(1): 126-133 (1994), "Affinity Isolation and
Characterization of Cytochrome P450 102 (BM-3) from Barbiturate-induced
Bacillus megaterium". I hope this helps you a bit.
= Shaun D. Black, PhD | Internet e-mail address: shaun at uthct.edu =
= Dept. of Biochemistry | University of Texas Health Center at Tyler =
= Tyler, TX 75710 | World Wide Web: http://psyche.uthct.edu =
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