The invention provides a preparation method of antibacterial anti-wrinkle linen-cotton fabric. The method comprises the following steps: (1) ultrasonically treating the linen-cotton fabric with an aqueous solution of triethanolamine; (2) preparing amino-modified silver nanoparticles; (3) modifying 2,3-epoxypropyltrimethyl ammonium chloride by a carbodiimide solution, then crosslinking with the amino-modified silver nanoparticles, aminated graphene, and chitosan, and drying the materials; and (4) adding a dried product obtained in the step (3), tea polyphenol, malic acid, tartaric acid, tripolymaleic acid, nano TiO2, antimony trioxide, an osmotic agent, and fibroin to an aqueous solution of ethanol to prepare a finishing agent, and performing dip padding on the fabric to obtain the antibacterial anti-wrinkle linen-cotton fabric. The linen-cotton fabric prepared by the invention has good inhibition effect on Escherichia coli, Staphylococcus aureus and Candida albicans, and has good anti-wrinkle performance. Therefore, the fabric made by the invention has broad application prospects as the antibacterial anti-wrinkle fabric.