This study aims to investigate the secondary metabolites produced by the Bacillus velezensis JS8.Four compounds were isolated from ISP2 liquid fermentation products of the B. velezensis JS8 by silica gel column chromatography and preparative high performance liquid chromatograph,and their structures were identified by the NMR and MS data as a new metabolite desmethyl venezuel G (1) and three known metabolites N-(5-formyl-2-hydroxyphenyl)acetamide (2),N-(2-hydroxy-5-methylphenyl)acetamide (3),and 3-acetamido-4-hydroxybenzoic acid (4) Based on the HPLC analysis of culture broths in different culture days,the biosynthetic pathway of compounds 1-4 was proposed.Compounds 1-4 were tested for antibacterial activities against Staphylococcus aureus and Escherichia coli.The results showed that only compound 3 had moderate antibacterial activity against S.aureus with minimum inhibitory concentration (MIC) value of 18.1 μg/mL.In addition,all compounds were evaluated for cytotoxicity against HeLa cell line and anti-inflammatory activity via inhibiting NO production in lipopolysaccharide-stimulated RAW 264.7 cells in vitro.The results indicated that compounds 1-4 showed no cytotoxic activity and no inhibitory effect against NO production with concentration of 50 μmol/L.