Fig. 2: Investigation of reductases NapW and homW in NDM biosynthesis. | Nature Communications

Fig. 2: Investigation of reductases NapW and homW in NDM biosynthesis.

From: Reductive inactivation of the hemiaminal pharmacophore for resistance against tetrahydroisoquinoline antibiotics

Fig. 2

a Genetic characterization in vivo. HPLC analysis with UV detected at 270 nm (I) wild-type S. lusitanus NRRL 8034; (II) mutant S. lusitanus TG3022 (ΔnapW); (III) mutant S. lusitanus TG3023 (ΔhomW); (IV) double mutant S. lusitanus TG3024 (ΔnapW&ΔhomW). b HPLC analysis of the enzymatic reaction products: full reaction catalyzed by NapW for 10 min using NADH (I) or NADPH (II) as cofactor; (III), full reaction mediated by homW; (IV), control reaction with heat-inactivated NapW; (V), 1 generated in situ by NapU from 2; (VI), standard of 2. c Chemical reactions by NapU and NapW/homW. d NDM resistance of S. lusitanus WT and the mutants. e Comparison of the antibacterial activity of NDM against E. coli BL21 (DE3) containing pET28a (vector), pTG3044 (napW), and pTG3045 (homW).

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