Monokaryotic pleurotus sapidus strains with intraspecific variability of an alkene cleaving dyp-type peroxidase activity as a result of gene mutation and differential gene expression
Creators: name: Krahe, Nina-Katharina
name: Berger, Ralf G.
name: Witt, Martin
name: Zorn, Holger
name: Omarini, Alejandra B.
name: Ersoy, Franziska
Publisher: name: Basel : MDPI
Year: 2021
Subjects: subject: Alkene cleavage
subject: Basidiomycota
subject: Biocatalysis
subject: Dikaryon
subject: Dye-decolorizing peroxidase (DyP)
subject: Gene expression
subject: Gene mutation
subject: Intraspecific variability
subject: Monokaryon
subject: Pleurotus sapidus
subject: alkene derivative
subject: peroxidase
subject: alkene
subject: coloring agent
subject: fungal protein
subject: recombinant protein
subject: transcriptome
subject: Article
subject: Basidiomycetes
subject: basidiospore
subject: biocatalysis
subject: comparative study
subject: conformational transition
subject: controlled study
subject: DNA sequence
subject: enzyme activity
subject: enzyme structure
subject: follow up
subject: fungal strain
subject: gene amplification
subject: gene expression
subject: gene mutation
subject: genetic variation
subject: heterologous expression
subject: Komagataella
subject: Komagataella phaffii
subject: Michaelis constant
subject: nonhuman
subject: nucleotide sequence
subject: Pleurotus
subject: protein stability
subject: real time polymerase chain reaction
subject: structural homology
subject: thermostability
subject: X ray crystallography
subject: biotransformation
subject: enzymology
subject: genetics
subject: metabolism
subject: molecular model
subject: mutation
subject: Alkenes
subject: Biotransformation
subject: Coloring Agents
subject: Fungal Proteins
subject: Models, Molecular
subject: Mutation
subject: Peroxidase
subject: Recombinant Proteins
subject: Transcriptome
subject: Dewey Decimal Classification::500 | Naturwissenschaften::570 | Biowissenschaften, Biologie
subject: Dewey Decimal Classification::500 | Naturwissenschaften::540 | Chemie

description: The basidiomycete Pleurotus sapidus produced a dye-decolorizing peroxidase (PsaPOX) with alkene cleavage activity, implying potential as a biocatalyst for the fragrance and flavor industry. To increase the activity, a daughter-generation of 101 basidiospore-derived monokaryons (MK) was used. After a pre-selection according to the growth rate, the activity analysis revealed a stable intraspecific variability of the strains regarding peroxidase and alkene cleavage activity of PsaPOX. Ten monokaryons reached activities up to 2.6-fold higher than the dikaryon, with MK16 showing the highest activity. Analysis of the PsaPOX gene identified three different enzyme variants. These were co-responsible for the observed differences in activities between strains as verified by heterologous expression in Komagataella phaffii. The mutation S371H in enzyme variant PsaPOX_high caused an activity increase alongside a higher protein stability, while the eleven mutations in variant PsaPOX_low resulted in an activity decrease, which was partially based on a shift of the pH optimum from 3.5 to 3.0. Transcriptional analysis revealed the increased expression of PsaPOX in MK16 as reason for the higher PsaPOX activity in comparison to other strains producing the same PsaPOX variant. Thus, different expression profiles, as well as enzyme variants, were identified as crucial factors for the intraspecific variability of the PsaPOX activity in the monokaryons. © 2021 by the authors. Licensee MDPI, Basel, Switzerland.