"The study aimed to estimate the influence of temperature, time, and ultrasound application during desizing and scouring of cotton with -amylase and pectinase, respectively, on the weight loss, breaking force and color difference between raw and bio-scoured cotton, to obtain purified and hydrophilic cotton, with a simultaneous slight decrease in mechanical properties. Under the optimal conditions, determined by the application of Response Surface Methodology (time – 30 min, temperature – 59.4 °C, with ultrasound treatment), a weight loss of 4.97%, color difference of 3.86, and a breaking force of 730.22N were obtained. The bio-scoured cotton fabric was also characterized in terms of electrokinetic and sorption properties, chemical composition and morphology of the cotton surface by zeta-potential measurement, wicking and contact angle determination, FTIR and SEM characterization, respectively. The developed enzymatic scouring process leads to obtaining purified, whiter and hydrophilic cotton, with slight changes in mechanical properties, which makes bio-scoured cotton fabric suitable for further wet processing."