Anoectochilus formosanus, a medicinal plant used to treat hypertension, lung disease, and liver disease, was grown to maximize biomass and secondary metabolite production in a controlled environment under four levels of photosynthetic photon flux (PPF), namely, 10, 30, 60, or 90 μmol·m−2·s−1, that is L10, L30, L60, and L90 treatments, respectively. On Day 45, all growth values were greatest for the L30 plants. Dry weight was lowest for the L10 plants. Leaf area, stem length, and fresh weight were lowest for the L90 plants. The chlorophyll concentration was highest in the L10 treatment and decreased with increasing PPF. Electron transport ratios of leaves were highest in the L30 treatment and lowest in the L90 for the second leaf (counted down from the apex) and in the L10 for the third leaf. An increase in light intensity from 10 to 60 μmol·m−2·s−1 increased the superoxide dismutase activity and was associated with an increase in the total flavonoid concentration. The total flavonoid concentration (mg·g−1 DW) was greatest in the L60 and lowest in the L90. However, the total flavonoid content (mg/plant) was highest in the L30 plants as a result of great biomass. The results indicated that A. formosanus is a typical shade plant suitable to grow under low light intensity at PPF of 30 to 50 μmol·m−2·s−1 for both growth and production of total flavonoid. A light intensity of 90 μmol·m−2·s−1 induced stress on plant growth and reduced photosynthetic capability and the flavonoid accumulation.