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Enhanced extracellular production of laccase in Coprinopsis cinerea by silencing chitinase gene

Summary

Scientists improved the production of laccase, an enzyme with industrial uses in detoxification and food processing, by genetically engineering mushroom cells to have stronger cell walls. By reducing the activity of genes that break down chitin in the cell wall, they created mushroom strains that could better withstand the stirring forces during fermentation, resulting in over twice as much enzyme production. This breakthrough could lead to cheaper, more efficient production of this useful green catalyst on an industrial scale.

Background

Laccase is a copper-containing polyphenol oxidase with important industrial applications as a green biocatalyst. While basidiomycetes produce over 90% of current laccase market share, their natural laccase yield is relatively low. Homologous recombinant expression in basidiomycetes can preserve enzymatic properties better than heterologous expression in ascomycetes.

Objective

This study aimed to enhance recombinant laccase Lcc5 extracellular production in Coprinopsis cinerea by silencing chitinase genes. The researchers identified critical chitinase genes through transcriptome analysis and used RNA interference to suppress their expression to improve laccase yield.

Results

Silencing ChiE2 reduced chitin content by 80%, decreased sporulation and growth rate, but increased cell wall shear tolerance. The engineered strain Cc lcc5-anti ChiE2-5 achieved maximum laccase activity of 38.2 U/mL at 250 rpm agitation with 0.35 U/mL·h productivity, 2.6-fold higher than the control strain at the same agitation speed.

Conclusion

This is the first report of enhanced recombinant laccase production in C. cinerea by chitinase gene silencing. The strategy effectively increased shear force tolerance and extracellular laccase yield, paving the way for industrial-scale laccase production and providing a novel approach for enhancing other target protein yields in basidiomycetes.
  • Published in:Applied Microbiology and Biotechnology,
  • Study Type:Experimental Research,
  • Source: PMID: 38713211, DOI: 10.1007/s00253-024-13164-9
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