Fig. 3: In vitro functional characterization of AlpH.
From: O-methyltransferase-like enzyme catalyzed diazo installation in polyketide biosynthesis
a HPLC analysis of the organic extracts from the enzymatic reactions: (i) DHR + AlpJ + AlpK + AlpH + NADH + gluN2H3; (ii) DHR + AlpJ + AlpK + AlpH + NADH; (iii) DHR + AlpJ + AlpK + boiled AlpH + NADH + gluN2H3; (iv) prekinamycin standard. (b) Direct HPLC analysis of the enzymatic reactions: (i) DHR + AlpJ + AlpK + AlpH + NADH + gluN2H3 (35 min of reaction time); (ii) DHR + AlpJ + AlpK + AlpH + NADH + gluN2H3 (120 min of reaction time); (iii) DHR + AlpJ + AlpK + AlpH + NADH; (iv) DHR + AlpJ + AlpK + boiled AlpH + NADH + gluN2H3; (v) prekinamycin standard. c LC-HRMS data (positive mode) of 5. d MS/MS (positive mode) spectrum and proposed fragmentation of 5.
