Transcriptional and enzymatic profiling of Pleurotus ostreatus laccase genes in submerged- and solid-state fermentation cultures

CASTANERA, RAUL; PÉREZ GÚMER; OMARINI, ALEJANDRA; ALFARO, MANUEL; PISABARRO, GERARDO; FARACO, VINCENZA ; AMORE, ANTONELLA; RAMIREZ, LUCIA

APPLIED AND ENVIRONMENTAL MICROBIOLOGY

AMER SOC MICROBIOLOGY

Lugar: Washington; Año: 2012 vol. 78 p. 4037 - 4045

0099-2240

The genome of the white rot basidiomycete Pleurotus ostreatus includes 12 phenol oxidase (laccase) genes. In this study, we examined their expression profile 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 used for the normalization and reaction efficiency determination) 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 (iSmF) cultures but downregulated in the solid (SSF) cultures, (ii) the Lacc2 and Lacc10 genes are the main source of laccase activity in the iSmF cultures upon induction with water-soluble wheat straw extracts, and (iii) an additional, as-yet-uncharacterized activity (Unk-1) is specifically induced in SSF cultures that complements the activity of Lacc2 and Lacc10. Moreover, both the enzymatic laccase activity and the Lacc gene family transcription profile greatly differ between closely related strains. These differences may be targeted for biotechnological breeding programs for enzyme production in submerged fermentation reactors.