pete with these guys...
Maybe someone else has other ideas about that?
Good luck!
Jan
--- Hari Namboodiri schrieb am Fr,
3.9.2010:
> Von: Hari Namboodiri
> Betreff: [ccp4bb] Heme Proteins
> An: CCP4BB@JISCMAIL.AC.UK
> Datum: Freitag, 3. September, 2010 19:30 Uhr
> Dear CCPeers
Dear CCPeers
I wanted to thank everyone for sharing their experiences and giving valuable
advice on CYP51 protein expression. It seems that special vectors like pCW-Lic
or pcWORi is needed for efficient expression. I have not found any commercial
source for them though. Could anybody be willing
At 5:56 PM +0100 8/26/10, Hari Namboodiri wrote:
Can anyone provide insights about expressing heme containing
proteins in E.col? Does E.coli need any porphyrin precursor during
expression or you need special E.coli strains. I have references
mentioning delta-aminolevulinic acid for one ortholo
: [ccp4bb] Heme Proteins
An: CCP4BB@JISCMAIL.AC.UK
Datum: Donnerstag, 26. August, 2010 18:56 Uhr
Hi CCPers
Can anyone provide insights about expressing heme containing proteins in E.col?
Does E.coli need any porphyrin precursor during expression or you need special
E.coli strains. I have references
Hari,
I have expressed sulfite oxidase (with a N-terminal heme) in E. coli strain
TP1000 which resulted in red protein. I did not supplement the media with iron
or a porphyrin precursor.
Sincerely,
James
On Aug 26, 2010, at 12:56 PM, Hari Namboodiri wrote:
> Hi CCPers
>
> Can anyone provi
I think you need a special trick to express C-type heme proteins.
For B hemes (protoheme, protoporphryn 9), which I think is
what the P450's have, E.coli is always expressing several
proteins which contain it. But I guess there may be some way
to up the production in case of an overexpresser.
Ha
Dear Hari,
You might look at Lucy Waskell's experiences with full length
mammalian Cytochrome P450 2B4. While she got >50 mg of purified
protein per liter of culture, many other heme proteins are much harder
to express in E. coli with heme assembly proteins, chaperones, etc.
You just ha
Hi CCPers
Can anyone provide insights about expressing heme containing proteins in E.col?
Does E.coli need any porphyrin precursor during expression or you need special
E.coli strains. I have references mentioning delta-aminolevulinic acid for one
ortholog but none for another. The enzyme is CY
