Usually you should add aminolevulinic acid to enhance the cytochrome production. L-glutamate is the starting step to produce 5-aminolevulinate using -HemA----HemL.
Hi Felix,
I am not aware of this being attempted before, and it could be interesting to test.
The one problem I can see with it would be that the cells would preferentially use the glutamate for other processes like peptide synthesis. Aminolevulinic acid is a direct precursor for porphyrin synthesis for heme or chlorophyll, and I am not aware of any other natural function for it. This therefore ushers the metabolism toward this production as the cells have clear 'instructions' to follow. In the case of glutamate, the question would be to what extent this pathway is switched on.
Naturally you would start with excess glutamate, so that the regular cellular requirements are met, then perhaps the increased demand caused by the recombinant cytochrome c550 would induce this pathway.
It would be interesting to see if it works.
Regards,
Chris
Hello Chris, thanks for your answer! I have to try to reduce costs....maybe I can find the way to activate the pathway. It was reported that glutamate enhances the denitrification in Bradyrhizobium (a lot of cytochrome cofactors in those enzymes), so maybe with a little help in microaerobic condition and adding a little bit of nitrate I can achieve the successful production! A big hug from Argentina. Next year I will be back at UFS for a short period of time.
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