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In this paper, a novel polysaccharide extracted from Inonotus obliquus, coding IOP, was prepared by water extraction and alcohol precipitation method, which was used to study the fermentation process of functional yoghurt supplemented with IOP (fermented IOP yoghurt). With sensory evaluation as the index, the preparation parameters of fermented IOP yoghurt, including IOP content, yoghurt starter content, sucrose content, fermentation time and fermentation temperature, were optimized by the single factor experiments and response surface methodology. Its quality and biological activities were compared with ordinary yoghurt without IOP and yoghurt with unfermented IOP. The results showed that the optimal technological parameters of fermented IOP yoghurt were 0.08% IOP, 4.45% yoghurt starter, 6% sucrose, fermentation time 4 h, fermentation temperature 42 ℃. Under this conditions, fermented IOP yoghurt possessed better sensory quality, nutritional value and texture properties, stronger DPPH radical and superoxide anion scavenging activities, more excellent diet-related α-glucosidase inhibitory activity. It was indicated that IOP had the potential application as a natural food additive in the development of function yoghurt due to the health benefits of IOP yoghurt. This study enriched the consumer market of yoghurt, helped to broaden the application of IOP in the food industry, and also opened up a new way for the development and utilization of the resources of Inonotus obliquus in China. [ABSTRACT FROM AUTHOR]