The invention discloses an elastin peptide with a beautifying effect as well as a preparation method and application thereof. The preparation method of the elastin peptide comprises the following steps: after pre-treating cattle neck ligaments, mixing the cattle neck ligaments with water; after adjusting the pH (Potential of Hydrogen) value, mixing the cattle neck ligaments with proteinase and carrying out enzymolysis; carrying out enzyme denaturing, centrifuging, filtering and collecting filtrate; taking the filtrate, concentrating and drying to obtain the elastin peptide. The method provided by the invention utilizes a biological enzymolysis technology, and a bioactive peptide with the beautifying effect is released from elastin of the cattle neck ligaments. The cattle neck ligament elastin peptide prepared by adopting the method has remarkable anti-wrinkle and moisturizing effects, can remarkably improve the moisture content in skins and the content of hyaluronic acid in blood serum and can also remarkably increase the content of collagen and the elastin in the skins.