I am NdmB, one of the enzymes encoded by the gene cluster of Pseudomonas putida CBB5. Together with my counterparts—NdmA, NdmC, NdmD, and NdmE—we function collaboratively to sequentially demethylate caffeine at the N1, N3, and N7 positions, ultimately synthesizing xanthine.
I am primarily responsible for the N3 demethylation of caffeine, but I struggle to catalyze this reaction using natural caffeine as a substrate to produce paraxanthine (PX), which frustrates me.
PX is my little golden bean friend, valued at $1522 per gram. Unfortunately, obtaining PX is not easy: natural extraction methods are challenging, and during chemical synthesis, halting the methylation reactions midway is difficult. As a result, some of my human friends are looking to me to produce PX through my demethylation reactions.
A group of friends from NEFU is eager to assist me. They plan to utilize directed evolution methods to facilitate my reunion with my little golden bean, PX, as soon as possible.
