RESUMEN
l-DOPA-2,3-dioxygenase from Streptomyces lincolnensis is a single domain type I extradiol dioxygenase of the vicinal oxygen chelate superfamily and catalyzes the second step in the metabolism of the propylhygric acid moiety of the antibiotic, lincomycin. In this report, the kinetic mechanism of l-DOPA dioxygenase is interrogated using stopped-flow in order to determine microscopic rate constants. Pre-steady state, progress curve and steady-state data were combined in a global kinetic analysis using KinTek Explorer in order to define and constrain a kinetic model for the type I l-DOPA dioxygenase. The data are best described by a four step mechanism, in which the cyclization of the enzymatic product is not enzyme catalyzed.
Asunto(s)
Oxigenasas/química , Streptomyces/enzimología , CinéticaRESUMEN
An automated solvent dispensing workstation capable of delivering volumes ranging from 10 mL to 4.5 L for the preparation of solutions/mobile phases was developed and implemented into the industrial R&D laboratory. The workstation was designed to address business, safety, and compliance needs while meeting or exceeding the precision and accuracy of current manual methods of preparation. The system's performance was optimized with respect to liquid transfer tubing inner diameter, pumping pressure, flow characteristics of the valve, and computer control logic. The automated solvent dispensing workstation was shown to exceed the specifications set by the ASTM for Class A graduated cylinders for all dispense volumes (10 mL-4.5 L).