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 com-pounds, including cordycepin, adenosine, and polysaccharides, that exhibit antioxidant, immunomodula-tory, and anticancer properties. Enhancing production and ensuring the genetic stability of these metabo-lites 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 mor-phology, 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 mor-phological 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 proper-ties, supporting its potential for large-scale pharmaceutical and nutraceutical applications.