This article is devoted to studying the musical and educational activities of the Yekaterinoslav Theological Seminary in the context of Russian spiritual education in the late nineteenth and early twentieth centuries. The Seminary's role in regional culture is considered in detail, with a specific focus on the development of music education and musical art. The extent of dissemination of musical knowledge by the Yekaterinoslav Theological Seminary is discussed. The role of Seminary musical education in preserving the traditions of church singing art is also studied. The connection between Seminary musical education and musical life is traced, as well as the emergence of secular musical art elements in the performance of sacred music in the concert repertoire of the Seminary choir, orchestra, and instrumental ensembles. Finally, the three most important functions of the Theological Seminary's musical and educational activities are formulated. This research is a contribution to the study of the process that supported Ukraine music education formation as a part of Europe.