Abstract

Ligninolytic heme peroxidases comprise an extensive family of enzymes, which production is characteristic for white-rot Basidiomycota. The majority of fungal heme peroxidases are encoded by multigene families that differentially express closely related proteins. Currently, there were very few attempts to characterize the complete multigene family of heme peroxidases in a single fungus. Here we are focusing on identification and characterization of peroxidase genes, which are transcribed and secreted by basidiomycete Trametes hirsuta 072, an efficient lignin degrader. The T. hirsuta genome contains 18 ligninolytic peroxidase genes encoding 9 putative lignin peroxidases (LiP), 7 putative short manganese peroxidases (MnP) and 2 putative versatile peroxidases (VP). Using ddPCR method we have quantified the absolute expression of the 18 peroxidase genes under different culture conditions and on different growth stages of basidiomycete. It was shown that only two genes (one MnP and one VP) were prevalently expressed as well as secreted into cultural broth under all conditions investigated. However their transcriptome and protein profiles differed in time depending on the effector used. The expression of other peroxidase genes revealed a significant variability, so one can propose the specific roles of these enzymes in fungal development and lifestyle.

Highlights

  • Heme peroxidases are large group of biocatalysts with ecological and biotechnological significance

  • While lignin peroxidase (LiP) is capable of unmediated oxidation of recalcitrant polymer, manganese peroxidase (MnP) act through mediated oxidation of phenolic intermediates [5]

  • LiPs are proposed to oxidize lignin with free radicals generated through oxidation of various secreted metabolites [6], and MnPs can produce reactive oxygen species by oxidation of fungal unsaturated fatty acids [7,8]

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Summary

Introduction

Heme peroxidases are large group of biocatalysts with ecological and biotechnological significance. We used the ddPCR method to quantify the absolute expression of all annotated peroxidase genes on different growth stages of basidiomycete, and during introduction of effectors (alkali lignin and bromocresol green dye) into cultural broth.

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