article · Notulae Scientia Biologicae
Multi-step mutagenesis was used to enhance laccase enzyme production in the fungus Lentinus edodes and apply it in the green synthesis of silver nanoparticles. Exposing the wild fungal strain to ultraviolet irradiation yielded a mutant, designated UV10, which generated 10.6 units per millilitre per minute of laccase after seven days of fermentation. This represented a fourfold yield improvement over the wild strain. Crude laccase from this mutant successfully mediated the rapid formation of walnut-shaped silver nanoparticles measuring between 50 and 100 nanometres, with fungal proteins driving the synthesis process. When evaluated against clinical bacterial isolates, the resulting nanoparticles demonstrated effective inhibition against Escherichia coli, Pseudomonas aeruginosa, and Klebsiella pneumoniae, highlighting a biological route for producing functional nanomaterials.
Biological synthesis offers an eco-friendly alternative to hazardous chemical routes for producing nanomaterials. Demonstrating that an improved fungal strain can generate high enzyme yields to produce silver nanoparticles that inhibit common clinical pathogens shows how fungal biotechnology can contribute to sustainable antimicrobial manufacturing and broader industrial enzyme processes.
The process could enable green manufacturing routes for antimicrobial nanoparticles, of interest to industrial biotechnology developers and producers of antimicrobial formulations. The work remains at an early laboratory stage, having validated strain improvement, particle synthesis, and in vitro antibacterial efficacy on a laboratory scale without establishing pilot-scale fermentation, synthesis yields, or formulation stability.
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This study reports the multi-step mutagenesis of Lentinus edodes towards optimization of the production of laccase and novel application of laccase in the biosynthesis of silver nanoparticles (AgNPs) which could be used to develop an eco-friendly method for the rapid biosynthesis of AgNPs. The wild strain of L. edodes was subjected to UV irradiation at 254 nm and the resultant viable mutant was further treated with acridine orange, a chemical mutagen. The strains were evaluated for the production of laccase and the crude laccase of the UV mutant (UV10) was used for the green synthesis of AgNPs. The particles were characterized by UV-Visible spectroscopy, Fourier transform infrared (FTIR) spectroscopy and scanning electron microscopy (SEM). Laccase activities of wild, UV10 and UV10ACR8 strains of L. edodes were obtained as 2.6, 10.6 and 2.8 U/ml/min respectively after 7 days of fermentation, showing laccase yield improvement of 4.08-fold for UV10 mutant. UV-Visible spectroscopy indicated the formation of AgNPs at absorption band of 430 nm. FTIR result indicated that proteins were responsible for AgNP synthesis, while SEM analysis confirmed the formation of walnut-shaped nanoparticles with size range of 50-100 nm. The biosynthesized nanoparticles revealed effective inhibition against clinical isolates of Escherichia coli, Pseudomonas aeruginosa and Klebsiella pneumoniae. To the best of the authors’ knowledge, this result represents the first report on the biosynthesis of AgNPs using L. edodes metabolite. The report adds to the growing relevance of L. edodes as potential industrially viable organism, used for diverse biotechnological applications.
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DOI: 10.15835/nsb749643
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