In recent years, the optimization of straw-rotting formulations for cultivating edible mushrooms and the management of the resulting spent mushroom substrate have emerged as new challenges. This study aimed to investigate the composting of spent mushroom substrate produced from mushroom cultivation with various straw additions, under conditions where chicken manure was also used. Parameters measured during the composting process included temperature, pH, electrical conductivity (EC), germination index (GI), moisture, and total nitrogen content. Additionally, changes in nutrient content within the compost piles before and after composting were determined, and the variations in bacterial and fungal communities across different treatments before and after composting were analyzed using 16S rRNA and ITS sequencing. The results indicated that the spent mushroom substrate produced by adding 20% straw during mushroom cultivation was more suitable for composting treatment. The findings suggest that incorporating an appropriate amount of straw in mushroom cultivation can facilitate subsequent composting of spent mushroom substrate, providing an effective strategy for both environmental protection and cost reduction.