Novel dyp-type peroxidases from actinobacteria: a genome mining approach
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Date
2024
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University of the Western Cape
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. only two of the constructs were successfully transformed: the dyp-type peroxidase genes from nocardia gamkensis czh20t and micromonospora sp. strain 30-1. further analysis using t7-primers confirmed sequence identity of the constructs.
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Keywords
Actinobacteria, Peroxidase, Dye-Decoloursing Peroxidase, Dyp-Type, Decolourisation