The chemical mechanism of the lysine monooxygenase Nocobactin G (NbtG) from Nocardia farcinica has been investigated using both steady-state and rapid reaction kinetics. NbtG is capable of hydroxylation of both L- and D-lysine. This less stringent substrate specificity indicates that the active site is different from related enzymes. O2 consumption is stimulated when both lysine stereoisomers are present indicating a form of regulation by lysine. Substrate does not affect the rate of reduction indicating that lysine stimulates a kinetic step in the oxidative half-reaction. Primary kinetic isotope effect studies indicate that NbtG stereospecifically transfers the pro-R hydride of NAD(P)H with a Dkred ~1.5. Solvent isotope studies yield a D2Okcat of ~2.8 indicating that an exchangeable proton step is rate limiting. This work was supported by a grant from the National Science Foundation MCB-1021384. 

Mechanistic studies on NbtG, a flavin dependent <i>N</i><sup>6</sup>-lysine monooxygenase

245th National Meeting (2013)

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Mechanistic studies on NbtG, a flavin dependent N6-lysine monooxygenase

Mechanistic studies on NbtG, a flavin dependent N⁶-lysine monooxygenase