Accumulated genome data of streptomycetes have revealed many peptide biosynthetic genes, but the number of reported peptides is relatively small. Recently, venezuelin was characterized via heterogeneous expression, as it is not produced in normal conditions. We thus conducted a chemical investigation on the extract of Streptomyces venezuelae to search for new peptides. We isolated a new hydrophobic peptide, named venepeptide, from the MeOH extract of S. venezuelae. Using matrix-assisted laser desorption/ionization–time-of-flight (MALDI-TOF) mass spectrometry, electrospray ionization Fourier-transform ion cyclotron resonance (ESI-FTCR) mass spectrometry, amino acid analysis, and nuclear magnetic resonance (NMR) spectra (1H, 13C, HMBC, HSQC, TOCSY, DQF-COSY, NOESY), we determined its molecular formula as C101H158N24O24S and its structure as an N-formylated 19-amino acid peptide (sequence: MNVITNLLAGVVHFLGWLV). From the whole-genome data of S. venezuelae, we identified the biosynthetic gene of venepeptide, which is flanked by genes SVEN_0784 (protein YceG) and SVEN_0785 (putative ABC transporter). A tblastn search revealed that analogous biosynthetic genes are distributed in six other streptomycetes, with 63% (12/19) amino acid identity among the expected analogs. Venepeptide has a high hydrophobic amino acid content (73%, 14/19 residues). Unlike the morphogenetic peptides SapB and SapT, venepeptide did not exhibit aerial hyphae formation activity in S. venezuelae. Additionally, it showed no antimicrobial activity against tested microorganisms (including Escherichia coli, Pseudomonas aeruginosa, Bacillus subtilis, Staphylococcus aureus, Saccharomyces cerevisiae, and Aspergillus niger) via the paper disc method.