Figure 8. PIZ Converts BRs to the corresponding acylated products in vitro.

(A) Structures of BL and ¹⁴C-lauric acid-CoA (LA-CoA). (B) Detection of ¹⁴C-labeled products extracted with ethyl acetate and separated by silica-gel thin layer chromatography. An asterisk marks the respective specific product for the applied substrates, BL, CS and TY, after 2 h of reaction at room temperature. An arrowhead marks free lauric acid (LA). e.v., empty vector control, n.s., no BR substrates. (C) LC-MS/MS analysis of the acylated products. Products were extracted from silica-gel after TLC and analysed on LC-MS/MS. Enzyme reactions were carried out using non-labelled LA and d₄-BL, d₄-CS or non-labelled TY. Predicted pseudo-molecular ions ([M+H]⁺) for the acylated products are shown with an asterisk.