Cloning and expression of versatile peroxidase from Rigidoporus vinctus

• Main Authors: Pan, Chun-Chen, 潘俊琛
• Other Authors: Lin, Chi-Tsai
• Format: Others
• Language: zh-TW
• Published: 2014
• Online Access: http://ndltd.ncl.edu.tw/handle/gfprcg

碩士 === 國立臺灣海洋大學 === 生物科技研究所 === 102 === Versatile peroxidase secreted by the white-rot fungus involved in the natural degradation of lignin. Versatile peroxidase can oxidize a variety of aromatic compounds by the catalytic tryptophan residue or Mn3+ which oxidized at the haem channel site. This lignin-degrading enzymes have potential applications in the pulp biobleaching, textile dyes decolorization, aromatic pollutants decomposition. In this study, a cDNA (1074 bp) encoding a putative VP from Rigidoporus vinctus was identified based on sequence homology. The deduced amino acid sequence (357 amino acids) is conseRved among the reported VP. The predicted molecular weight of RvVP was 34.8 kDa. A 3-D structural model of RvVP was constructed based on the crystal structure of Pleurotus eryngii VP (PDB code 2VKA). The active sites for oxidation of Mn2+ ions and for oxidation of aromatic substrates were identified, both characteristic for versatile peroxidase. To characterize the RvVP, the coding region was subcloned into an expression vector pET-20b(+) and pYEX-S1, then transformed into Escherichia coli and Saccharomyces cerevisiae. The recombinant RvVP expressed as inclusion bodies in E. coli and the soluble recombinant RvVP was obtained in S. cerevisiae, but the catalytically inactive this might be ascribable to absence of heme group.