Built in the 6th and 7th centuries during the Baekje period, the Buyeo Royal Tombs consist of seven tombs, including Tomb No. 1, which contains murals. To preserve Tomb No. 1 from damage caused by microorganisms, periodic microbial-distribution investigations are conducted. Following such investigations in August 2016, June 2018, and November 2019, the microbes were classified according to the investigation period, location of collection, and space. This study compares and analyzes the results. The concentration of airborne microorganisms in Tomb No. 1 and the number of microbial genera identified in each space of the tomb decreased as proximity to the main room with murals diminished. During the investigation period, the genera <i>Bacillus, Cladosporium, Penicillium,</i> and <i>Streptomyces</i> were commonly identified on Tomb No. 1. The microorganisms collected f rom the main room walls were mostly isolated from the east and west walls where the genera <i>Bacillus, Cupriavidus, Paenibacillus, Pseudomonas,</i> and <i>Streptomyces</i> were commonly identified in three or more walls. In particular, the genus <i>Streptomyces</i> is a dangerous strain capable of damaging murals by penetrating into and discoloring the pigments on them. The data generated from this study may be useful for future research on microbial distribution in other domestic mural tombs and those located in North Korea and abroad.