In the present study, an attempt has been made to investigate the bio-bleaching preparation of Egyptian cotton fibers. Bleaching of the fibers was performed with hydrogen peroxide, which was enzymatically produced by glucose oxidase during oxidation of D-glucose. The results obtained were compared with the conventional processing method. Promising results have been reported by Glucose Oxidase enzyme (Gox) that converts glucose to hydrogen peroxide and gluconic acid used for bleaching purpose. The gluconic acid produced is a good chelator and can be used as a sequestering agent for metal ions. This is so-called Indirect Enzymatic Bleaching Process and the whiteness degree obtained was acceptable compared with conventional chemical method. The mechanical properties such as tensile strength and elongation of cotton fibers showed no major influence using Gox compared with the conventional method.