Fig. 1

The role of CYP79 and CYP83 in glucosinolate pathway. The first two steps of glucosinolate biosynthesis are catalysed by P450 enzymes from CYP79 and CYP83 families that convert amino acids into oximes and further into nitrile oxides, which can be conjugated with glutathione by a glutathione-S-transferase (GST) or spontaneously. We measured the generation of phenylalanine-derived products to evaluate the effect of N-terminal sequence modifications on P450 functionality in E. coli: phenylacetaldoxime (Phe-Ox), when engineering CYP79A2 alone, and S-phenylacetohydroxymoyl-L-glutathione (Phe-GSH), when engineering CYP79A2 together with CYP83s