Cucumber (Cucumis sativus L.) is substantial economic importance in South Korea, with its cultivation occupying a significant portion of agricultural land. However, sustainable cucumber production requires effective management of various diseases affecting the crop yield. In this study, we explored the potential of Streptomyces sp. S20-465-derived metabolites in controlling cucumber anthracnose disease caused by Colletotrichum orbiculare. This study identified 4-hydroxy-4-methyl-2-pentanone as a potent antifungal compound present in the n-hexane extract of Streptomyces sp. S20-465 culture filtrate. This compound exhibited significant the disease reduction, demonstrating their potential as control agents. Our findings suggest that Streptomyces sp. S20-465-derived metabolite could serve as an effective tool for managing cucumber anthracnose, offering a sustainable approach to enhancing cucumber production. Further research into the application and efficacy of these compounds in agricultural system will be conducted.