Cloning,expression and characterization of a feruloyl esterase C from Penicillium chrysogenum

Objective:To clone feruloyl esterase gene C from Penicillium chrysogenum and characterize the general properties of the enzyme.Methods:The feruloyl esterase C gene was amplified by PCR based on the Penicillium chrysogenum feruloyl esterase C gene sequence and cloned into the expression vector p PIC9K,resulting the recombinant plasmid p PIC9K-Pcfae C.The recombiant plasmid was linerized and transformed into P.pastoris by electroporation.The transformants was screened based on the transparent zone technology.The screened transformants was then induced by methanol.the enzymatic properties of the protein were then measured.Results:SDS-PAGE analysis showed that the molecular mass of the enzyme was about 30 k D.The length of the gene was 762 bp.It comprised one open reading framwork(ORF)and annotated to encode 249 amino acid.The optimal temperature and p H was found to be 40℃and 6,respectively.Moreover,the recombinant enzyme was stable at 40-50℃and p H 5-7.Conclusion:The enzyme successfully expressed in P.pastoris could laid theoretical foundation in food,fodder and paper making industry.