Synthetic fibers are used extensively in textile industry, however, their high hydrophobicity is a drawback that needs to be considered. The decrease of hydrophobicity can be achieved via a ‘green” root using enzymes as biocatalysts. In this study, the enzymatic surface modification of polyamide (PA) 6.6 fabric was studied with the use of the commercial protease Alcalase 2.4 L at optimal conditions. The modified fabrics were studied via dyeing parameters K/S and ΔΕ values. For treatment at 40–60 °C and pH 8 ΔE was found to be approximately 14 and K/S was 1.24-fold increased. Additionally, the enzymatic surface modification of PA textile was justified using different spectroscopy techniques, such as FTIR-ATR and XPS. FTIR-ATR indicated alterations of Cdouble bond; length as m-dashO and N-H band intensities, while via XPS, there proved to be differences in relative intensities of carbon component peaks. Finally, thermogravimetric and mechanical tests were also conducted to prove the non-degradation of the properties of the bulk material. In conclusion, the investigated enzymatic process increased the hydrophilicity with 2.7-fold increased water absorbency and 1.24-fold enhanced color strength of PA textiles, while maintaining the thermal and mechanical properties of the bulk synthetic material.