The purpose of this study was to investigate the extraction yield and physicochemical characteristics of mucin from the epidermal mucus of Misgurnus mizolepis. Ninety percent ethanol produced the highest yield of mucin. At higher concentrations, mucin had more elastic properties, as revealed by dynamic viscoelasticity measurements. Differential scanning calorimetry (DSC) demonstrated that mucin is a thermostable glycoprotein. Denaturation enthalpy increased as mucin concentration increased. Mucin inhibited the growth of both Escherichia coli and Bacillus cereus, but had better antimicrobial activity in the former. Mucin also exhibited antioxidant activity, equivalent to 66.64% of that of L-cysteine. Mucin inhibited elastase activity and MMP-1 & MMP-2 expression activity, and induced hyaluronic acid expression. It is indicating that mucin is effective in moisturizing and anti-aging the skin. Thus, mucin from the epidermal mucus of Misgurnus mizolepis could have great potential as a non-food material in various industries.