Abstract: Among many plant polysaccharides, arabinogalactan had the best growth-promoting effect on Akkermansia muciniphila (Akk) model strain (DSM 22959). In order to further explore the influence of arabinogalactan on Akk metabolites, arabinogalactan was used as part of the carbon source to ferment Akk in vitro for the first time, and determined the supplementary amount of arabinogalactan to facilitate the Akk fermentation broth. In the subsequent analysis of the samples, high-resolution non-targeted metabolomics technology was used to detect and analyze the differential metabolites in the Akk fermentation broth samples. The results showed that there were 85 different metabolites (VIP>1, P<0.05) and 32 different metabolic pathways. This study confirmed that the arabinogalactan could affect metabolic pathways, biosynthesis of amino acids, 2-oxocarboxylic acid metabolism, ABC transporters and other metabolic pathways, promoting Akk to produce more metabolites with special effects, such as hypoxanthine, mohanone phenol, propionic acid, etc., which had potential of weight loss, anti-cancer and blood pressure lowering, respectively. The results of this study will provide a theoretical basis for the use of arabinogalactan as a prebiotics of Akk or its biogenesis in the future.