Facile One-Pot Immobilization of a Novel Esterase and Its Application in Cinnamyl Acetate Synthesis

Cinnamyl acetate has a wide application in food industries. This work focuses on cinnamyl acetate synthesis with a novel esterase. est GUZ753 , an esterase gene from Geobacillus uzenensis DSMZ 13551, was first cloned and expressed in Pichia pastoris KM71. The optimal activity of Est GUZ753 towards p -NP caprylate was at pH 8.0 and 70 °C, and the half-life at 70 °C was 28 h. Furthermore, Est GUZ753 showed marked tolerance in methanol. The activity of Est GUZ753 increased up to 1.16-fold in 90% methanol after 72 h, and the half-life in it was 336 h. The crude fermentation broth of Est GUZ753 was immobilized directly onto the epoxy resin (Lx-105s), and the immobilized Est GUZ753 exhibited a 99% conversion for cinnamyl acetate synthesis at a high cinnamyl alcohol concentration in 6 h (cinnamyl alcohol: 1.0 M, enzyme dosage: 3%). These characteristics of Est GUZ753 indicated its great potential for organic synthesis. Graphic Abstract A novel esterase gene from Geobacillus uzenensis DSMZ 13551 was first cloned and expressed in Pichia pastoris KM71. Esterase Est GUZ753 showed marked thermostability at high temperature and tolerance in methanol. Furthermore, the crude fermentation broth of Est GUZ753 was immobilized directly onto the epoxy resin, and immobilized Est GUZ753 exhibited a 99% conversion for cinnamyl acetate synthesis.