Down fiber – a natural and environmentally recyclable insulation material – is applied mainly in the area of natural-filled products. However, owing to its natural deficiencies of low fluffiness and thermal stability, large-scale application is limited. In our work, an enzyme preparation (transglutaminase (TGase)) was used as a fluffy agent to improve the fluffy degree of down. With fluffy as the main index, the single factor condition of only using TGase enzyme was optimized, and fluffiness was increased by 22%. On the basis of this optimum condition, the compound dosage of papain and TGase was further optimized. The fluffy degree of down was improved by hydrolysis of papain and crosslinking of TGase. Amino group content, thermogravimetry (TG), differential scanning calorimetry (DSC), moisture absorption, tensile strength, and elongation at break of down treated with different enzyme reagents were tested. The results showed that the thermal stability of down fiber treated with the multi enzyme increased significantly, the denaturation temperature was increased from 53.7°C to 77.4°C, and the moisture absorption was also improved. The elastic property also exhibited a great enhancement; elongation at break increased from 12.4% to 37.7%, nearly three times higher than the original property, and the tensile strength increased from 180 MPa to 370 MPa, almost 2.1 times as much as before, increased by 105.6%.