SummaryAs the flexitarian diet becomes more popular, as well as rising concerns about the environment and health wellness, the demand for plant‐based meat has significantly increased in recent years. This study aimed to develop the feasibility of using mushrooms as an alternative ingredient in a plant‐based meat analogue via high‐moisture twin‐screw extrusion. Five high‐moisture fibrous‐structured meat analogues (HMs, moisture around 61–64 wt, %) from soy protein isolate (SPI) mixed with mushroom powder (MP) in the ratios of SPI and MP of 60:0 (control: CT), 45:15 (M15), 35:25 (M25), 25:35 (M35), and 15:45 (M45) were successfully developed. The increase in MP led to a significant change in darker and brown spot scattering attributes; however, it significantly decreased the textural properties (hardness, and chewiness) of the selected HM (M25), which were significantly lower than CT (P < 0.05). Water holding capacity slightly decreased, while oil holding capacity decreased in HMs with MP compared to CT. Low protein solubility and viscosity of HMs proved the thermally denatured when extrusion cooking. The protein content of all HMs ranges from 52.29 to 72.14 wt% in concordance with the increment of the SPI ratio. HMs showed higher protein digestibility during the intestinal stage than the gastric stage. Thai‐Northern style sausage (Sai‐aua) analogue prepared from M25 gained the highest overall acceptability. Due to their availability, low cost, nutritional values, and health benefits, mushrooms proved to be a potential source to produce HMs. Besides, our findings help create high‐added‐value products that benefit human health, protein sustainability, and environmental concerns.