Expression of Pleurotus eryngii versatile peroxidase in Escherichia coli and optimisation of in vitro folding

Heterologous expression of Pleurotus eryngii versatile peroxidase (VP) in Escherichia coli was investigated. The cDNA encoding the mature sequence of the allelic variant VPL2 was cloned into the expression vector pFLAG1 and expressed in E. coli W3110. After induction with isopropyl- beta -D-thiogalactopyranoside (IPTG), the recombinant polypeptide (VPL2*) was found to be the major protein located in inclusion bodies. In vitro folding of VPL2* was initially performed under conditions previously determined for folding of recombinant lignin peroxidase from Phanerochaete chrysosporium (LiPH8*) but a very low yield of active enzyme was obtained (