Journal article

Publication year: 2017

Pages: 164-

Journal: AMB Express

Volume number: 7

ISSN: 2191-0855

Open access status: Gold

DOI Link: https://doi.org/10.1186/s13568-017-0463-5

Publisher: SpringerOpen

Abstract: 
The coding sequence of a peroxidase from the secretome of Pleurotus sapidus was cloned from a cDNA library. Bioinformatic analyses revealed an open reading frame of 1551 bp corresponding to a primary translation product of 516 amino acids. The DyP-type peroxidase was heterologously produced in Trichoderma reesei with an activity of 55,000 U L-1. The enzyme was purified from the culture supernatant, biochemically characterized and the kinetic parameters were determined. The enzyme has an N-terminal signal peptide composed of 62 amino acids. Analysis by Blue Native PAGE and activity staining with ABTS, as well as gel filtration chromatography showed the native dimeric state of the enzyme (115 kDa). Analysis of the substrate range revealed that the recombinant enzyme catalyzes, in addition to the conversion of some classic peroxidase substrates such as 2,2'-azino-bis(3-ethylthiazoline-6-sulfonate) and substituted phenols like 2,6-dimethoxyphenol, also the decolorization of the anthraquinonic dye Reactive Blue 5. The enzyme also catalyzes bleaching of natural colorants such as beta-carotene and annatto. Surprisingly, beta-carotene was transformed in the presence and absence of H2O2 by rPsaDyP, however enzyme activity was increased by the addition of H2O2. This indicates that the rPsaDyP has an oxidase function in addition to a peroxidase activity. As a consequence of the high affinity to the characteristic substrate Reactive Blue 5 the rPsaDyP belongs functionally to the dyp-type peroxidase family.

Harvard Citation style: Lauber, C., Schwarz, T., Nguyen, Q., Lorenz, P., Lochnit, G. and Zorn, H. (2017) Identification, heterologous expression and characterization of a dye-decolorizing peroxidase of Pleurotus sapidus, AMB Express, 7, p. 164. https://doi.org/10.1186/s13568-017-0463-5

APA Citation style: Lauber, C., Schwarz, T., Nguyen, Q., Lorenz, P., Lochnit, G., & Zorn, H. (2017). Identification, heterologous expression and characterization of a dye-decolorizing peroxidase of Pleurotus sapidus. AMB Express. 7, 164. https://doi.org/10.1186/s13568-017-0463-5