FAD in culture medium?

Georg Wille moqch at mlucom6.urz.uni-halle.de
Fri Nov 13 03:41:16 EST 1998


Hi all,

I'm about to express a recombinant protein in E.coli that requires FAD
as a cofactor, otherwise it doesn't fold correctly - at least in vitro.

I'm concerned that if protein expression (T7 promoter) is faster than
FAD synthesis I'll end up with large amounts of denatured protein,
inclusion bodies possibly - as has happened already in a similar
experiment. Those are very, very difficult to refold in my case so I'd
like to avoid them.

Do you have any experience whether additional FAD in the culture medium
or some cheaper precursor like FMN or riboflavin would be beneficial
here? As far as I know E.coli can make FAD from "scratch", but can it
utilize one of the mentioned precursors more efficiently?


Thanks a lot!

Georg


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