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

Summary

Scientists enhanced the production of laccase, a useful enzyme with industrial applications, in a type of mushroom by silencing specific genes involved in cell wall construction. The modified mushroom strain could withstand stronger mixing forces during fermentation, leading to significantly higher enzyme yields. This genetic engineering approach could help make laccase production more efficient and cost-effective for industrial uses like detoxification and food processing.

Background

Laccase is a copper-containing polyphenol oxidase with important industrial applications as a green biocatalyst. Basidiomycetes laccases are highly valued but have relatively low natural yields. Homologous recombinant expression in Coprinopsis cinerea can preserve the enzymatic properties of basidiomycetes laccase better than heterologous expression.

Objective

To enhance the extracellular production of laccase Lcc5 in C. cinerea by silencing critical chitinase genes using RNA interference. The study aimed to investigate how chitinase gene silencing affects mycelial morphology and recombinant laccase yield.

Results

ChiE2 silencing reduced sporulation and growth rate but increased shear force tolerance and extracellular laccase activity to 38.2 U/mL at 250 rpm in the 3-L fermenter, 2.6-fold higher than control strain. Silenced strains demonstrated increased cell wall roughness and Young’s modulus, enabling survival at higher agitation speeds.

Conclusion

Silencing the chitinase gene ChiE2 successfully enhanced recombinant laccase production in C. cinerea by improving tolerance to shear forces during fermentation. This novel strategy provides a pathway for scaling-up Lcc5 production and establishing C. cinerea as a high-expression system for laccase production.
  • Published in:Applied Microbiology and Biotechnology,
  • Study Type:Experimental Research,
  • Source: PMID: 38713211, DOI: 10.1007/s00253-024-13164-9
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