This study examined the use of cutinase and lipase to process cotton/polyester blend fabric. Optimum treatment conditions for cutinase and lipase were investigated for cotton/polyester blend fabric. The properties of enzyme-treated fabrics were evaluated and compared in optimal treatment conditions. In addition, the possibility to provide an enzymatic finishing on blend fabrics using mixed enzymes in a two-step process were studied. The weight loss of cotton/polyester blend fabrics with Triton X-100 was 0.8% and the dyeing property of blend fabrics with calcium chloride increased by a factor of 1.2. The use of two enzymes in combination with cutinase and lipase in the presence of auxiliaries resulted in a cotton/polyester blend fabric weight loss of 0.8%. In addition, the dyeing properties of cotton/polyester blend fabrics improved by a factor of 1.5 and the moisture regain of cotton/polyester blend fabrics improved by a factor of 1.16. However, no marked loss was observed in tensile strength. The surface morphology of cotton/polyester blend fabrics is modified through a two-enzyme treatment. The treatment of cotton/polyester blend fabrics with cutinase and lipase maintains cotton strength and improves the moisture regain of polyester fabrics.