Streptomycetes are known for biosynthesizing a wide variety of structurally complex secondary metabolites, and genome data have revealed numerous peptide biosynthetic genes, though the number of reported peptides is relatively few. Recently, the new lantibiotic peptide venezuelin was characterized via heterogeneous expression of biosynthetic genes based on genome mining. In this context, we conducted a chemical investigation on the extract of Streptomyces venezuelae to search for new peptides. As a result, we isolated a new hydrophobic peptide named venepeptide from the MeOH extract of S. venezuelae. Structural determination using amino acid analysis, cyanogen bromide (CNBr) treatment, N-terminal protein sequencing, matrix-assisted laser desorption/ionization–time-of-flight (MALDI-TOF) MS/MS, and NMR spectroscopy (1H, 13C, HMBC, HSQC, TOCSY, DQF-COSY, NOESY) showed that venepeptide is an N-formylated 19-amino acid peptide with the sequence MNVITNLLAGVVHFLGWLV and molecular formula C101H158N24O24S. Genome search using the tblastn program identified the venepeptide biosynthetic gene in S. venezuelae, which is flanked by genes SVEN_0784 (protein YceG) and SVEN_0785 (putative ABC transporter). Additionally, analogous biosynthetic genes were found in six other streptomycetes, with 63% (12/19) of amino acids being identical among the expected peptide analogs and conserved hydrophobic regions. Venepeptide has a high hydrophobic amino acid content (73%, 14/19 residues). Activity assays revealed that unlike SapB and SapT, venepeptide did not show aerial hyphae formation activity on S. venezuelae. Furthermore, it did not exhibit any antimicrobial activity against tested microorganisms (including Escherichia coli, Pseudomonas aeruginosa, Bacillus subtilis, Staphylococcus aureus, etc.) via the paper disc method.