Induction of laccase in Basidiomycetes: the laccase-coding messenger

Area of Science:

• Molecular biology
• Mycology
• Enzymology

Background:

• Laccases are enzymes with diverse industrial applications.
• Understanding the regulation of laccase gene expression is crucial for optimizing their production.
• Fungal laccases are particularly well-studied due to their varied metabolic capabilities.

Purpose of the Study:

• To investigate the formation and characteristics of mRNA encoding inducible laccase in selected fungal species.
• To determine the stability (half-life) of this specific mRNA.
• To confirm the translation of this mRNA into functional laccase protein.

Main Methods:

• Extraction and detection of mRNA from fungal mycelia treated with ferulic acid.
• Measurement of mRNA half-life using techniques to assess RNA stability.
• Analysis of mRNA molecular weight.
• In vitro synthesis of polypeptides using polysomes from treated mycelia and comparison with laccase protein.

Main Results:

• Specific mRNA for inducible laccase was successfully identified in Coriolus versicolor, Pleurotus ostreatus, and Pholiota mutabilis.
• The half-lives of these laccase-specific mRNAs were determined to be 30, 37, and 24 minutes, respectively.
• The molecular weight of the newly synthesized mRNA in Pleurotus ostreatus was approximately 4.5x10^5, aligning with the size of the inducible laccase protein.
• In vitro translation experiments using polysomes from ferulic acid-treated mycelium produced a polypeptide with electrophoretic mobility identical to that of laccase.

Conclusions:

• The study provides evidence for the formation of specific mRNA molecules that direct the synthesis of inducible laccase in fungi.
• The determined mRNA half-lives offer insights into the regulation and turnover of laccase gene expression.
• The results confirm that the identified mRNA is translated into laccase protein, validating its role in enzyme production.