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Cordyceps militaris (CM) is a critical fungus for promoting health, while the cordycepin has been recognized as an indicator compound with bioactive functions such as induction of apoptosis, anti-inflammation, inhibition of angiogenesis, and antioxidant properties. However, the cultivation of CM required appropriate light to induce the fruiting body into regular growth. Light-emitting diodes (LED), which combine different wavelengths and conserve energy, offer promising potential for applications in CM cultivation. Therefore, this study involved using low-power and low-illumination LED light sources of different wavelengths for intervention in the photoperiod of CM cultures to observe the effects on the growth pattern, bioactive substance content, and antioxidant activity. This study showed that the growth patterns of CM developed normally when irradiated with low-brightness LED light sources of different wavelengths. Moreover, the total phenolic content (TPC), cordycepin, and relative antioxidant activity levels in CM fruiting body water extract were significantly elevated upon exposure irradiated with a glare blue light wavelength range compared to other groups (p < .05). Hence, this study proved a high potential for LED irradiation to be applicable in CM industrial development and exploration of practical knowledge in industrial biotechnology with facilitated CM products that can be economically produced.