Cytochrome P450 BM3

[Shaun D. Black]

Greetings,

     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.


Best regards,

Shaun Black


  =-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=
  = 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  =
  =-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=