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Cloning and Characterization of a Novel Laccase Gene, fvlac7, Based on the Genomic Sequence of Flammulina velutipes

Summary

This research identified and characterized a new enzyme-producing gene in the winter mushroom Flammulina velutipes. The enzyme, called laccase, is important because it can break down various compounds and has potential applications in multiple industries. Impact on everyday life: • Could help develop more environmentally friendly methods for paper production • May contribute to better waste water treatment solutions • Could lead to improved industrial processes for breaking down harmful pollutants • Potential applications in development of biosensors for medical and environmental monitoring • May help create more efficient and sustainable industrial processes

Background

Laccases are copper-containing polyphenol oxidases found in white-rot fungi, higher plants, insects and bacteria. They are particularly abundant in white-rot fungi involved in lignin metabolism. These enzymes catalyze the reduction of oxygen to water while oxidizing aromatic compounds, with applications ranging from effluent treatment to pulp bleaching and biosensors.

Objective

To investigate the molecular characteristics and expression of a novel laccase gene (fvlac7) from the Flammulina velutipes strain 4019-20 using genomic information. The study aimed to clone and analyze the nucleotide sequence of this new laccase gene based on the fungal genomic sequence.

Results

The study identified a novel laccase gene fvlac7 with a 1,608 bp open reading frame encoding 538 amino acids. The genomic DNA sequence contained 16 introns. The protein showed only 42-51% identity with 12 different mushroom species’ laccases, including two from F. velutipes. The protein contained a 25-amino acid signal sequence, four copper-binding sites, and four N-glycosylation sites. When expressed in E. coli, it produced a protein of approximately 60 kDa.

Conclusion

The fvlac7 represents a novel laccase gene from F. velutipes with distinct molecular characteristics from previously known laccases. While the gene was successfully expressed in E. coli, the protein could not be purified in an active form, suggesting the need for alternative expression systems to study its enzymatic properties.
  • Published in:Mycobiology,
  • Study Type:Laboratory Research,
  • Source: 10.5941/MYCO.2013.41.1.37
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