The biosynthesis of ZnO is performed through enzymatic mechanisms for controlling the particle size and morphology with using the sol‐gel method. Furthermore, this method utilizes a stabilizer obtained from the cell biomass of Aspergillus niger mold in order to yield homogeneous and consistent products. The evaluation of functional, morphological, and antibacterial activities was carried out at pH 6.0‐13.0. The analysis FT‐IR showed the interaction of hydroxyl groups, aromatic rings, as well as N−H and O−Zn‐O compounds at a wavenumber of 401–584 cm−1. The XRD and SEM characterizations showed that ZnO structure and crystal phase were hexagonal wurtzite at dimensions of 36.2 ‐ 45.4 nm. The differences in pH also influenced the dimensions, morphology, and antimicrobial activity. ZnO with a pH of 8.0 was characterized by FESEM‐EDAX, based on the analysis of morphological uniformity. This characterization obtained rod and cube structures, with atomic ratios of Zn=61.5 % and O=38.5 %. The UV‐DRS spectrum showed that the optical band gap with a value of Eg=3.00–3.11 eV. The differences in morphology further distinguished antibacterial properties on textile fibers, through the use of the Gram ‐ and + bacteria (Pseudomonas aeruginosa and Staphylococcus aureus), with inhibition zones of 17–21 mm and 21–25 mm, respectively. Therefore, ZnO is classified as a very strong antibacterial material than amoxicillin, with an inhibition zone of 13.6 mm.