Vespa crabro and V. analis are social hornet species commonly found in Asia, including Korea. Mastoparan is one of the major venom peptides of these two hornets but its amino acid sequence defers substantially. To examine the differences in the potential toxicity and bioactivity of mastoparans between these two social hornets, differential toxicological and pharmacological activities of synthesized mastoparan were investigated. V. analis mastoparan showed a 7-fold higher hemolytic activity, suggesting its higher cytotoxic potential compared with V. crabro mastoparan. Mastoparans from both hornet species exhibited similar levels of antimicrobial activities against Staphylococcus aureus and Botrytis cinerea, whereas the mastoparan from V. analis showed more potent antimicrobial activities against Escherichia coli and Candida albicans. Nevertheless, the antimicrobial activities of mastoparans of V. crabro and V. analis were relatively lower compared with those of other wasps. Both mastoparans also exhibited some levels of antitumor activity but the activity was significantly higher in V. analis mastoparan. In summary, the hemolytic, antimicrobial, and antitumor activities of synthesized V. analis mastoparan were higher than those of V. crabro mastoparan. These differential bioactivities are likely due to the amino acid sequence differences in the mature peptides. In particular, the additional Lys residue present in V. analis mastoparan may contribute to the higher levels of bioactivity as proposed by secondary structure prediction.