<jats:title>Abstract</jats:title><jats:p>Two novel cyclic hexapeptides, designated NW‐G08 (<jats:bold>1</jats:bold>) and NW‐G09 (<jats:bold>2</jats:bold>), were isolated from the fermentation broth of <jats:italic>Streptomyces alboflavus</jats:italic> 313. Their structures were elucidated on the basis of extensive spectroscopic analysis, MS experiments, and chemical analysis. Their antibacterial activities against several strains of bacteria were evaluated by micro‐broth dilution method. NW‐G08 (<jats:bold>1</jats:bold>) and NW‐G09 (<jats:bold>2</jats:bold>) were highly antibacterial against <jats:italic>Gram</jats:italic>‐positive but not active against <jats:italic>Gram</jats:italic>‐negative bactaria. The minimum inhibitory concentrations (<jats:italic>MIC</jats:italic>s) of <jats:bold>1</jats:bold> against <jats:italic>Bacillus cereus, Bacillus subtilis</jats:italic>, and <jats:italic>Staphylococcus aureus</jats:italic> were 6.25, 6.25, and 1.56 μg/ml, respectively. It is worth noting that <jats:bold>1</jats:bold> and <jats:bold>2</jats:bold> exhibited much higher activities against methicillin‐resistant <jats:italic>Staphylococcus aureus</jats:italic> (MRSA) than ampicillin, which implies that they might be potential candidates for the development of novel drugs against resistant pathogenic bacteria.