title: Transcriptional and enzymatic profiling of Pleurotus ostreatus laccase genes in submerged and solid-state fermentation cultures.
abstract: The genome of the white rot basidiomycete Pleurotus ostreatus includes 12 phenol oxidase (laccase) genes. In this study, we examined their expression profiles in different fungal strains under different culture conditions (submerged and solid cultures) and in the presence of a wheat straw extract, which was used as an inducer of the laccase gene family. We used a reverse transcription-quantitative PCR (RT-qPCR)-based approach and focused on determining the reaction parameters (in particular, the reference gene set for the normalization and reaction efficiency determinations) used to achieve an accurate estimation of the relative gene expression values. The results suggested that (i) laccase gene transcription is upregulated in the induced submerged fermentation (iSmF) cultures but downregulated in the solid fermentation (SSF) cultures, (ii) the Lacc2 and Lacc10 genes are the main sources of laccase activity in the iSmF cultures upon induction with water-soluble wheat straw extracts, and (iii) an additional, as-yet-uncharacterized activity (Unk1) is specifically induced in SSF cultures that complements the activity of Lacc2 and Lacc10. Moreover, both the enzymatic laccase activities and the Lacc gene family transcription profiles greatly differ between closely related strains. These differences can be targeted for biotechnological breeding programs for enzyme production in submerged fermentation reactors.
authors.affiliation: Department of Agrarian Production, Public University of Navarre, Pamplona, Spain.
authors.forename: Raúl
authors.initials: R
authors.lastname: Castanera
authors.forename: Gúmer
authors.initials: G
authors.lastname: Pérez
authors.forename: Alejandra
authors.initials: A
authors.lastname: Omarini
authors.forename: Manuel
authors.initials: M
authors.lastname: Alfaro
authors.forename: Antonio G
authors.initials: AG
authors.lastname: Pisabarro
authors.forename: Vincenza
authors.initials: V
authors.lastname: Faraco
authors.forename: Antonella
authors.initials: A
authors.lastname: Amore
authors.forename: Lucía
authors.initials: L
authors.lastname: Ramírez
mesh_terms: D001709:Biotechnology / Q000379:methods; D003470:Culture Media; D005285:Fermentation; D005656:Fungal Proteins / Q000235:genetics / Q000378:metabolism; D020869:Gene Expression Profiling; D015971:Gene Expression Regulation, Enzymologic; D015966:Gene Expression Regulation, Fungal; D042845:Laccase / Q000235:genetics / Q000378:metabolism*; D009172:Mycology / Q000379:methods; D020076:Pleurotus / Q000201:enzymology* / Q000235:genetics / Q000254:growth & development*; D020133:Reverse Transcriptase Polymerase Chain Reaction
chemical_list: D003470:Culture Media; D005656:Fungal Proteins; D042845:Laccase