Heterologous expression and physicochemical characterization of a fungal dye-decolorizing peroxidase from Auricularia auricula-judae

Characterization and application of a novel class II thermophilic peroxidase from Myceliophthora thermophila in biosynthesis of polycatechol

“…The Soret band appeared at 407 nm, and the addition of excess hydrogen peroxide led to a shift of the band to 420 nm, and to the formation of an extra band at 557 nm, as previously reported for Basidiomycetous heme-containing peroxidases [45,53].…”

“…K m values of MtPerII calculated in the present work are similar to VP (VPL2) from Pleurotus eryngii (0.54 mM for ABTS and 0.051 mM for H 2 O 2 ; [7], wild-type dye-decolorizing peroxidase DyP from Auricularia auricula-judae (283 M for ABTS; [45]) and VP (BadVP) from Bjerkandera adusta (17 M for H 2 O 2 with ABTS; [52]), among others. In terms of catalytic efficiency, MtPerII exhibits a k cat /K m value, for both ABTS and H 2 O 2 (401.2 mM −1 s −1 and 6470.1 mM −1 s −1 , respectively), comparable to the enzymes reported above, such as VPL2 from P. eryngii (410 mM −1 s −1 for ABTS and 2650 mM −1 s −1 for H 2 O 2 ; [7]), DyP from A. auricula-judae (2300 mM −1 s −1 for ABTS; [45]), BadVP from B. adusta (3,4 × 10 6 s −1 M −1 for H 2 O 2 with ABTS; [52]) and Coprinopsis cinerea peroxidase CiP (1,6 × 10 6 s −1 M −1 ; [52]). …”

“…Similar studies report that the half-lives of peroxidases in the presence of excess H 2 O 2 lie in the minute range [2,7,45], reaching up to 60 min, even after directed evolution approaches for the improvement of their properties. MtPerII appears relatively stable after 24 h in the presence of 5000-fold excess peroxide.…”

“…P. pastoris has been the host of choice for the expression of many eukaryotic proteins, due to its many advantages, such as protein processing, protein folding, and post-translational modifications [23]. On the other hand, E. coli hosts have been used for peroxidases expression [2,43], but the recombinant enzyme seems to be produced in inclusion bodies [44,45], creating the need for extra purification steps and/or in vitro folding of the protein [6].…”

Computational Modeling Methods for Understanding the Interaction of Lignin and Its Derivatives with Oxidoreductases as Biocatalysts

“…The structural difference of them with the other class of fungal, plant and animal peroxidases is that they do not have well conserved amino acids around the heme moiety. Initial studies in DyPs exhaustively characterized this region and after being reported the cloning and expression of Auricularia auricula-judae DyP (AauDyP) in E. coli [84], studies of the substrate oxidation sites in AauDyP were reported. In this context, computational studies have helped to obtain a better characterization of AauDyP and to give molecular details in the different applications that have been explored in this family of peroxidases.…”