ShLysG is a novel g-type lysozyme isolated from Hippocampus abdominalis in 2016. However, its antibacterial, antioxidant and anti-inflammatory properties have not been investigated. In this study, a recombinant ShLysG (rShLysG) fused with a 6xHis tag was produced in Escherichia coli with barely any hemolytic activity or cytotoxicity. It exhibited a broad antimicrobial spectrum against gram-positive and gram-negative bacteria with minimum inhibitory concentration (MIC) values between 20 and 60 mu g/mL. The bioactivity of rShLysG was stable in high temperature and extreme acid base conditions. In addition, rShLysG was resistant to papain and pepsin, and it showed a concentration-dependent antioxidant activity. By inhibiting the phosphorylation of NF-kappa B, the inflammatory cytokines expression level, including TNF-alpha, IL-6, and IL-8 in RAW264.7 macrophage cells induced by LPS were decreased. Taken together, rShLysG is a promising substitute to antibiotics as an antimicrobial and immunoregulatory pharmaceutical peptide for clinical use.