Dear all,
Sorry for non-crystallography related question.
We are trying to express an eukaryotic heme protein in E. coli. We are able to
express it in soluble form. When we use 5-Aminolevulinic Acid, E. coli becomes
brown. However, after cell lysis the soluble protein contains no heme and the
pellet is brown. If we extract the pellet with detergent (Triton X-100 and
Tween-20) the color comes in the supernatant, but there is no protein of our
interest. These eventually indicate that the protein and heme are getting
expressed/synthesized, but heme is not getting incorporated in the expressed
protein. We would like to get this protein in heme-bound holo-form.
Any suggestion to trouble-shoot this problem would be highly appreciated.
Replies can be sent to me directly and I will post a summary on a later date.
Thanks in advance,
Sincerely yours,
Biswajit Pal
CCMB, Hyderabad, India
