HPLC and LC-MS/MS Analysis of Reaction Products of ChLAC8 with Different Monolignols.
(A) to (F) HPLC chromatograms of reactions of ChLAC8 with caffeyl alcohol ([A] and [B], peak1), coniferyl alcohol ([C] and [D], peak 2), and sinapyl alcohol ([E] and [F], peak 3). (A), (C), and (E) show reactions incubated without recombinant enzyme. (B), (D), and (F) show reactions 30 min after incubation with recombinant ChLAC8.
(G) to (L) EICs, scanned at m/z = 329 for the dimer of caffeyl alcohol ([H], peak 5), m/z = 357 for the dimer of coniferyl alcohol ([I] and [J]), or m/z = 417 for the dimer of sinapyl alcohol (L, peak 7). (—) Indicates control assays without ChLAC8. No dimers were detected in the corresponding control reactions ([G], peak 4; [I] and [K], peak 6) or the reaction of ChLAC8 toward coniferyl alcohol (J).