The influence of light exposure on antioxidant and antimicrobial activities of nine fungal isolates [Pseudopestalotiopsis theae (EF13), Fusarium solani (EF5), Xylaria venustula (PH22), Fusarium proliferatum (CCH), Colletotrichum boninese (PL9), Colletotrichum boninese (PL1), Colletotrichum boninese (OL2), Colletotrichum gloeosporioides (OL3) and Colletotrichum siamense (PL3)] were determined. The isolates were incubated in blue, red, green, yellow and white fluorescent light (12h photoperiod of alternating light/dark). It was observed that green light induced higher total phenolic content (TPC) (2.96 ± 0.16mg-30.71 ± 1.03mg GAE/g) and ferric reducing antioxidant power (FRAP) in most isolates (4.82 ± 0.04-53.55 ± 4.33mg GAE/g), whereas red light induced higher total flavonoid content (TFC) levels (1.14 ± 0.08-18.40 ± 1.12mg QE/g). The crude extracts from most fungal cultures exposed to green and red lights were also notably more potent against the tested pathogens, as larger zones of inhibition (ZOI) (9.00 ± 1.00-38.30 ± 2.90mm) and lower minimum inhibitory concentration (MIC) (0.0196-1.25mg/mL) were achieved for antimicrobial effect. This study showed that light treatments are effective strategies in enhancing production of more potent antimicrobial compounds and valuable antioxidants from fungal isolates.