The valorization of olive mill wastewaters (OMWW), a by-product of the olive milling, is getting rising attention. Lipophilization of the main phenolic compound 3-hydroxytyrosol (HT) could facilitate its extraction. An immobilized variant of the promiscuous hydrolase/acyltransferase from Pyrobaculum calidifontis VA1 (PestE) was used to perform acetylation in water using ethyl acetate as acyl donor. PestE was used in a segmented flow setting to allow continuous operation. Additionally, HT precursors were made accessible by pretreatment with almond beta-glucosidase and the hydrolytic activity of PestE_I208A_L209F_N288A.