“…E. coli strains JM109, BL21(DE3), and the expression vector pETDuet-1, were obtained from Novagen (Madison, USA). The plasmid pETDuet-cut and pET24a-cut opt , which harbor the native and optimized genes encoding T. fusca cutinase respectively, were constructed and stocked in our previous work (Su et al, 2015(Su et al, , 2013b. The pMD18-T simple vector, DNA polymerase, restriction enzymes, alkaline phosphatase, and T 4 DNA ligase were obtained from Takara (Dalian, China).…”