Expression a Cutinase from Fusarium oxysporum in Pichia pastoris and Characterization

A 693 bp gene and encodes 231 mature amino acids for cutinase from Fusarium moniliforme Sheld was cloned and constructed into pPIC9K plasmid.The recombinant cutinase was hyper-produced extracellularly using Pichia pastoris GS115 as the host.After induction in a shake flask for 96 h, the yield of recombinant cutinase reached 71.68 U/mL, and the enzyme purified displayed a specific activity of 2490.1 U/mg.The optimum pH was 9.0, and the activity of the recombinant cutinase was relatively stable within the range of pH5.0~9.0.The optimal temperature of this enzyme was 35℃, and more than 70% of relative activity was retained after incubation for 1 h at 40℃. The recombinant enzyme activity could be stimulated by concentration of KCl, Triton X-100, MnCl₂, SDS, but inhibited by NaCl, BaCl₂, CuSO₄, Tween-20, FeSO₄, ZnSO₄, NiCl₂, EDTA, Tween-80.