Abstract: To investigate the deep-processing suitability of the same Lentinus edodes cultivar ('Shenxiang 1513') from different geographical origins (Suizhou, Hubei and Qingyuan, Zhejiang, China), aqueous extracts were prepared using three distinct extraction methods: hot water extraction, ultrasonic-assisted hot water extraction, and microwave-assisted hot water extraction (designated as HH, HU, HM for Hubei samples and ZH, ZU, ZM for Zhejiang samples, respectively). The physicochemical properties and structural characteristics of these extracts were systematically analyzed using high-performance liquid chromatography, ultraviolet spectroscopy, and fourier-transform infrared spectroscopy. The results demonstrated that the polysaccharide content of the six aqueous extracts ranged from 37.44% to 53.17%, while the protein content ranged from 14.70% to 27.03%. Notably, both HH and ZH exhibited higher polysaccharide and protein contents compared to the other extraction methods. Furthermore, the protein content of Zhejiang-derived extracts consistently surpassed that of the extracts from Hubei. Additionally, HU and ZM showed relatively higher ash contents among all tested samples. FTIR spectra confirmed the presence of polysaccharide-specific peaks (O-H, C-H) and protein-specific peaks (N-H) in all extracts. Amino acid analysis revealed complete profiles with glutamic acid dominant, ZM had the highest total amino acid content while HH had the lowest, with HU showed elevated individual amino acid levels compared to HH and HM, while ZM surpassed ZH and ZU. Molecular weight analysis revealed a trimodal distribution for the water extract of Hubei-sourced L. edodes, while the Zhejiang-sourced extract exhibited a bimodal distribution. Monosaccharide composition analysis demonstrated that all extracts consisted of mannose, glucosamine hydrochloride, galactose, and glucose, with glucose content exceeded 90%, both ultrasonic and microwave treatments significantly increased the monosaccharide content. In conclusion, significant differences in physicochemical and structural properties were observed among the aqueous extracts of L. edodes from different regions under varying extraction methods, for optimal extraction of bioactive compounds from aqueous extracts, hot water extraction proved to be the most effective method for both geographical origins. However, to obtain extracts with higher amino acid content, ultrasonic-assisted hot water extraction was preferable for Hubei-derived samples, while microwave-assisted hot water extraction yielded superior results for Zhejiang-sourced specimens. These findings provide a theoretical foundation for evaluating the deep-processing suitability of L. edodes and support the precise development of future health foods using region-specific extracts as functional ingredients.