To prevent and improve various metabolic-related diseases caused by modern high-energy eating habits, alternative meats using mushroom materials are being researched. In this study, high-moisture (HMMA) and low-moisture meat analog (LMMA) were prepared using Pleurotus ostreatus fruiting body (oyster mushroom) powder and isolated soy protein as the raw materials in a co-rotating twin-screw extruder. Textural characteristics tended to decrease as the oyster mushroom content increased. HMMA exhibited a fibrous structure similar to that of chicken, whereas LMMA did not show a characteristic fibrous structure. The water absorption capacity of substitute meat decreased with increasing mushroom powder content. Radical scavenging activity, a measure of antioxidant activity, increased with increasing mushroom content in the substitute meat because of the influence of antioxidant components such as polyphenols in mushrooms. In terms of the prepared substitute meat's color, it was less vibrant and lacked intensity, which is thought to make it less appealing to customers. To address this issue, more ingredients need to be investigated.