High-performance liquid chromatography of carbapenem antibiotics in complex biological samples with column switching and simultaneous multichannel ultraviolet monitoring

Non-Heme Iron Oxygenases Generate Natural Structural Diversity in Carbapenem Antibiotics

“…In Vitro reactions in MOPS, pH 7.0, containing Fe(NH 4 ) 2 (SO 4 ) 2 , R-KG, ascorbate, the subject carbapenam/em, and either ThnG or ThnQ were incubated and analyzed by HPLC for the formation of new product(s). 17,18 Clear outcomes were observed for both ThnG-and ThnQ-catalyzed reactions with PS-5 (5) (Figure 2 and Supporting Information). The products were immediately identifiable as carbapenems by their unique chromophores (λ max ) 290-320 nm).…”

“…The recombinant proteins were over-produced in E. coli Rosetta2(DE3) and purified by Ni-NTA affinity chromatography. In vitro reactions in MOPS, pH 7.0, containing Fe(NH 4 ) 2 (SO 4 ) 2 , α-KG, ascorbate, the subject carbapenam/em and either ThnG or ThnQ were incubated and analyzed by HPLC for the formation of new product(s) 17,18. Clear outcomes were observed for both ThnG- and ThnQ-catalyzed reactions with PS-5 ( 5 ) (Figure 2 and Supporting Information).…”