We have successfully identified the transcription factorCmr1from the fungusAureobasidium pullulansHit-lcy3T, which regulates melanin biosynthesis genes. Bioinformatics analysis revealed that the Cmr1 gene encodes a protein of 945 amino acids, containing two Cys2His2zinc finger domains and a Zn(II)2Cys6binuclear cluster domain located at the N-terminus ofCmr1. To investigate the function of theCmr1gene, we performed gene knockout and overexpression experiments. Our results showed thatCmr1is a key regulator of melanin synthesis in Hit-lcy3T, and its absence caused developmental defects. Conversely, overexpression ofCmr1significantly increased the number of chlamydospores in Hit-lcy3Tand improved melanin production. RT-qPCR analysis further revealed that overexpression of Cmr1 enhanced the expression of several genes involved in melanin biosynthesis, includingCmr1,PKS,SCD1, andTHR1. Melanin extracted from the Hit-lcy3Twas characterized using UV and IR spectroscopy. Furthermore, we assessed the antioxidant properties of Hit-lcy3Tmelanin and found that it possesses strong scavenging activity against DPPH·, ABTS·, and OH·, but weaker activity against O2−·. These findings suggest that Hit-lcy3Tmelanin holds promise for future development as a functional food additive.