Structural studies reveal flexible roof of active site responsible for ω-transaminase CrmG overcoming by-product inhibition

摘要

a Scheme for CrmG-catalyzed reaction. CrmG reaction pathway consists of oxidative deamination of an amino donor, resulting in by-product and PMP formation, and reductive amination of amino acceptor CRM M, leading to CRM N production, and PLP regeneration. b Nonlinear regression analysis of the competitive inhibition of α-ketoglutarate. c Nonlinear regression analysis of the competitive inhibition of pyruvate. d CrmG reaction progress with 1.5 equivalent amino donor. The reaction was performed in 50 μL assay solution (50 mM Tris pH7.5, 0.1 M NaCl, and 5% Glycerol) containing 4 μM CrmG, 80 μM PLP, 0.3 mM amino donor (l-Glu, l-Gln, or l-Ala), and 0.2 mM CRM M, in triplicates.