A DyP-Type Peroxidase of Pleurotus sapidus with Alkene Cleaving Activity

• Published in: Molecules (Basel, Switzerland) Vol. 25; no. 7; p. 1536
• Main Authors: Krahe, Nina-Katharina, Berger, Ralf G, Ersoy, Franziska
• Format: Journal Article
• Language: English
• Published: Switzerland MDPI AG 27.03.2020; MDPI
• Subjects: Aldehydes; Aldehydes - chemistry; alkene cleavage; Alkenes; Alkenes - chemistry; Amino acid sequence; Amino acids; Anethole; Anisoles - chemistry; Annatto; Anthraquinone; Anthraquinone dyes; Anthraquinones - chemistry; Aromatic compounds; aryl alkenes; basidiomycota; beta Carotene - metabolism; Biocatalysis; Biotransformation; Bixaceae - metabolism; Bleaching; Bleaching Agents - chemistry; Bleaching Agents - metabolism; Carotene; carotene degradation; Carotenoids - metabolism; Cleavage; Coloring Agents - chemistry; Decoloring; dye-decolorizing peroxidase (DyP); Dyes; Enzymes; Flavor; Flavors; Gene Expression; Hydrogen peroxide; Hydrogen Peroxide - chemistry; Hydrogen Peroxide - metabolism; Hydrogen-Ion Concentration; Isoeugenol; Ligands; Manganese - chemistry; Olfactory properties; Oxidation; Oxidation-Reduction; Peptides; Peroxidase; Peroxidase - chemistry; Peroxidase - isolation & purification; Peroxidase - metabolism; Plant Extracts - metabolism; Pleurotus - enzymology; Pleurotus - metabolism; Protein structure; Proteins; Saccharomycetales - metabolism; Styrenes - chemistry; Tertiary structure; β-Carotene; carotene degradation; Pleurotus sapidus; alkene cleavage; aryl alkenes; Komagataella pfaffii; basidiomycota; dye-decolorizing peroxidase (DyP); manganese; biocatalysis
• ISSN: 1420-3049; 1420-3049
• DOI: 10.3390/molecules25071536

Alkene cleavage is a possibility to generate aldehydes with olfactory properties for the fragrance and flavor industry. A dye-decolorizing peroxidase (DyP) of the basidiomycete (PsaPOX) cleaved the aryl alkene -anethole. The PsaPOX was semi-purified from the mycelium via FPLC, and the corresponding gene was identified. The amino acid sequence as well as the predicted tertiary structure showed typical characteristics of DyPs as well as a non-canonical Mn -oxidation site on its surface. The gene was expressed in GS115 yielding activities up to 142 U/L using 2,2'-azino-bis(3-ethylbenzthiazoline-6-sulphonic acid) as substrate. PsaPOX exhibited optima at pH 3.5 and 40 °C and showed highest peroxidase activity in the presence of 100 µM H O and 25 mM Mn . PsaPOX lacked the typical activity of DyPs towards anthraquinone dyes, but oxidized Mn to Mn . In addition, bleaching of -carotene and annatto was observed. Biotransformation experiments verified the alkene cleavage activity towards the aryl alkenes ( )-methyl isoeugenol, -methylstyrene, and -anethole, which was increased almost twofold in the presence of Mn . The resultant aldehydes are olfactants used in the fragrance and flavor industry. PsaPOX is the first described DyP with alkene cleavage activity towards aryl alkenes and showed potential as biocatalyst for flavor production.