Optimization of Recombinant Cutinase Fermentation Conditions and Its Application in Ethyl Acetate Synthesis

Abstract:

In order to achieve efficient production of the cutinase from Sirococcus conigenus，the optimization of fermentation conditions was investigated in P. pastoris KM71 harboring the plasmid pPIC9K-ScCuA. The optimized fermentation conditions were as follows：the inducing temperature is 28 ℃，the induction phase methanol concentration is maintained at 1% When the initial cell concentration OD600 reached 100. Under the optimum condition，the enzyme activity reached 419 U/mL at cultivation of 138 h，was 2 times than before.Different reaction parameters for the synthesis of ethyl acetate by direct esterification in organic solvents are investigated. The highest yield of 94.12% is obtained at 0.1 mol/L acetic acid，0.15 mol/L alcohol using a reaction temperature of 50 ℃ for 16 h incubation time in iso-octane.