This case report study investigated the botanical, physicochemical, and phenolic compositions, as well as the antioxidant and antimicrobial properties of varying amounts of black cumin honey (Nigella sativa) from Burdur region of Türkiye. The samples had an average moisture value of 17.23% and a Nigella sativa pollen ratio ranging from 19% to 51%. Electrical conductivity (EC) values ranged from 0.36 to 0.54 mS/cm. The average color values of Hunter (L, a, b) were 43.55 (L*), 32.77 (a*), and 64.52 (b*), respectively. The proline contents were ranged from 875 to 1624 mg/kg. The average F/G ratio and F+G value were 1.43±0.11 and 68.75±4.75, respectively. The average of total phenolic (TP) and total flavonoid (TF) contents were 36.87±7.00 mg GAE/100 g and 5.63±1.45 mg QUE/100 g, respectively. The antioxidant capacities of the samples were tested using the ferric reducing/antioxidant power (FRAP) and 2,2-Diphenyl-1-picrylhydrazyl (DPPH) assays, with mean values of 179.40±25.35 μmol FeSO4.7H2O/100 g and 46.07±12.87 mg/mL, respectively. The phenolic composition of the samples were tested using the HPLC-PDA method with 25 phenolic standards. Gallic acid, p-OH benzoic acid, t-cinnamic acid, p-coumaric acid, chrysin, and pinocembrin were determined as common components. The samples exhibited high antimicrobial activities against Staphylococcus aureus, Escherichia coli, Chromobacterium violaceum, Bacillus cereus, Klebsiella pneumoniae, and Acinetobacter haemolyticus, but did not affect Candida albicans, Candida parapilopsis, or Mycobacterium smegmatis.