The present study aimed to investigate the influence of ripening on the physicochemical, microbiological aspects, and fatty acid profile of Artisanal Coalho Cheeses and to detect if there are peptides with bioactive potential in their composition. Artisanal Coalho Cheese samples were kindly provided by a dairy farm located in Brazil in the Rio Grande do Norte state. A completely randomized design was adopted, with four maturation periods (0, 30, 45, and 60 days). Physicochemical traits (pH, total solids, moisture, non-fat solids, fat in total solids, protein, ash, fatty acid profile) and microbiological characterization (Salmonella sp, Listeria monocytogenes, total and thermotolerant coliforms, Staphylococcus aureus) were analyzed on cheese samples. Additionally, assays were performed for antioxidant and antihypertensive bioactivity through ACE and antimicrobial inhibition of the peptides extracted from the samples. There was a linear increase in total solids and ash content and a decrease in moisture content with increasing maturation time. The matured cheese samples had a lower pH than fresh Artisanal Coalho Cheese. Twenty-seven fatty acids were identified in the cheeses: 15 saturated, 07 monounsaturated, and 05 polyunsaturated, with a linear reduction of essential fatty acids (n6 and n3) during maturation. The microbiological quality of the cheeses was satisfactory, with an absence of undesirable bacteria in 92% of the cheese samples. Water-soluble peptide fractions from all periods tested showed antioxidant and antihypertensive potential with ACE control, and the maturation process potentiated these capacities, with a decline in these activities observed at 60 days. The antimicrobial activity against Gram-positive and Gram-negative bacteria increased with maturation, reaching better results until 60 days. The maturation process on wooden planks in the periods of 30, 45, and 60 days allows the production of Artisanal Coalho Cheese of an innovative character, safe to consumers from the microbiological point of view, with differentiated physicochemical and functional characteristics and good quality of lipid fraction compared to fresh cheese, enabling the addition of value to the dairy chain.