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. estGUZ753, an esterase gene from Geobacillus uzenensis DSMZ 13551, was first cloned and expressed in Pichia pastoris KM71. The optimal activity of EstGUZ753 towards p-NP caprylate was at pH 8.0 and 70 °C, and the half-life at 70 °C was 28 h. Furthermore, EstGUZ753 showed marked tolerance in methanol. The activity of EstGUZ753 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 EstGUZ753 was immobilized directly onto the epoxy resin (Lx-105s), and the immobilized EstGUZ753 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 EstGUZ753 indicated its great potential for organic synthesis. A novel esterase gene from Geobacillus uzenensis DSMZ 13551 was first cloned and expressed in Pichia pastoris KM71. Esterase EstGUZ753 showed marked thermostability at high temperature and tolerance in methanol. Furthermore, the crude fermentation broth of EstGUZ753 was immobilized directly onto the epoxy resin, and immobilized EstGUZ753 exhibited a 99% conversion for cinnamyl acetate synthesis.