Strain Evaluation and Multi-Generational Stability of Cordyceps militaris for Functional Bioactive Compound Production

BACKGROUND: Cordyceps militaris is a medicinal fungus known for producing bioactive compounds, including cordycepin, adenosine, and polysaccharides, that exhibit antioxidant, immunomodulatory, and anticancer properties. Enhancing production and ensuring the genetic stability of these metabolites are essential for therapeutic development. OBJECTIVE: To evaluate growth characteristics, metabolite yield, antioxidant activity, and genetic stability of 20 C. militaris strains, and to identify high-performing, stable candidates suitable for pharmaceutical applications. METHODS: Twenty C. militaris strains were cultured and analyzed for colony diameter, fruiting body morphology, biomass yield, and bioactive compound content. Antioxidant activity was assessed using the DPPH assay. Genetic stability was monitored over five generations via MAT gene profiling. RESULTS: Strain KTDT8 showed superior performance with the highest levels of cordycepin (469.8 ± 16.4 mg/100 g), adenosine (121.8 ± 7.4 mg/100 g), and polysaccharides (37.0 ± 2.9 mg/g), and the strongest antioxidant activity (IC₅₀ = 31.2 μg/mL). The strain maintained morphological and genetic stability across four generations, with a moderate decline in the fifth. DISCUSSION: KTDT8 consistently demonstrated superior metabolite yield, antioxidant capacity, and morphological stability across generations. Despite a moderate decline in biomass and fruiting body quality in the fifth generation, the strain retained its core phenotypic and genetic traits. These findings support KTDT8 as a viable candidate for long-term cultivation and bioactive compound production. CONCLUSION: KTDT8 represents a genetically stable, high-yielding strain with strong antioxidant properties, supporting its potential for large-scale pharmaceutical and nutraceutical applications.