FAD containing Protein purification?

17 April 2024 1 7K Report

I am trying to express a FAD containing enzyme of mycobacteria in E.coli. I am able to purify the protein which is slightly yellow in color but it seems that my FAD is all unbound to the protein. How can I express the protein which has tightly bound FAD?

Adam B Shapiro

You can try simply adding FAD back to the purified enzyme. It is commercially available.

https://www.sigmaaldrich.com/US/en/substance/flavinadeninedinucleotidedisodiumsalthydrate82951anhydrousbasis84366814

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