Silk fibroin (SF) hydrogel has been popular in the wound dressing and tissue engineering fields for its good biocompatibility, biodegradation and water retention ability. The development of functional SF‐based hydrogels has gained attention in recent decades in medical fields. Multifunctional SF hydrogel is prepared by the addition and combination of therapeutic agents with cell growth promoting, antibacterial, antioxidant and anti‐inflammatory properties. In particular, the antibacterial activity of SF‐based hydrogel is essential in clinical practice. Despite this growing interest in functional silk protein hydrogels and their potential antimicrobial applications, a comprehensive review on this topic is currently lacking. Therefore, this review aims to provide an in‐depth understanding of the advanced status of the antibacterial application of SF‐based hydrogel, mainly focused on different components loaded. The review begins by briefly introducing the preparation methods of SF‐based hydrogel. Subsequently, the review summarizes diverse components loaded into the hydrogel to provide a comprehensive understanding in the antibacterial field. Furthermore, the article reviews the biomedical applications of antibacterial aspects including tissue engineering, sustained drug release, wound repair and adhesive use. Also the challenges associated with developing SF‐based hydrogels are analyzed alongside future prospects. We hope that this study will contribute to promoting the application and innovation of SF material in the clinical antibacterial field. © 2024 Society of Chemical Industry.