Novel DyP-type peroxidases from actinobacteria: a genome mining approach

Abstract

Twenty-one whole genome actinobacterial sequences were obtained from strains previously isolated from various environments. A genome-mining approach was applied to identify the presence of Dye-decolourising (DyP-type or DyPs) peroxidases. From genome sequence annotation twenty-six DyP-type sequences were identified and through bioinformatic analysis for the presence of twin-arginine translocation (Tat) pathway signal peptides, was classified as belonging to class I (formerly class A) bacterial DyP-type peroxidases. Phylogenetic analysis of the DyP sequences showed that the Streptomyces derived DyPs were predominantly grouping together and the nonStreptomyces derived DyPs followed the same pattern. Eight DyP-type peroxidases were selected for cloning, with the majority being derived from Streptomyces spp., and primers were designed to amplify the DyP-type peroxidase genes. The PCR-amplicons were subjected to restriction digests and were ligated into pET20b(+). The constructs were transformed into Escherichia coli JM109.