This study aimed to select Perenniporia fraxinea isolates that form mycelial mats with excellent texture and ultimately to use them to produce mycelial based leather (MBL). Among 10 P. fraxinea isolates, JK-M0825 had the fastest mycelial growth rate on malt extract-yeast extract glucose medium. Polymorphic DNA of P. fraxinea isolates was detected using ultra-fast photonic polymerase chain reaction, which showed DNA diversity. The ribosomal DNA-internal transcribed spacer sequence of JK-M0825 was 99% identical to that of P. fraxinea isolates registered in GenBank. Four P. fraxinea isolates formed high-quality mycelial mats on sawdust substrates, and JK-M0825 produced the thickest mycelial mat (3.8 mm). JK-M0825 had the highest tensile strength (6.08 N/mm2) and an elongation rate of 32.93±6.31%. In addition, the mycelia of JK-M0825 formed uniformly on a large-scale culture medium (90 cm x 90 cm) and formed a textured mycelial mat. These results suggest that JK-M0825 is appropriate for producing MBL.