α-Glucosidase Inhibitory Activity of Polyphenol-rich Sugarcane extract: Screening and Mechanistic Insights Based on Biolayer Interferometry-Mass Spectrometry

Mengli Yao¹Jia Liu¹Fang Zhou¹Haizhi Li¹Ruoyong Wang²ZHONG HAN³Jie Liu⁴Wei Chen⁵Guoyu Liu¹Shuheng Yang¹Shenlin Duan¹Xiaofeng Han^1*Peng Yuan^1*

• ¹China National Research Institute of Food and Fermentation, Beijing, China
• ²Air Force General Hospital PLA, Beijing, Beijing Municipality, China
• ³School of food science and engineering, South China University of Technology, Guangzhou, Guangdong Province, China
• ⁴Key Laboratory of Geriatric Nutrition and Health, Beijing Technology and Business University, Beijing, Beijing Municipality, China
• ⁵Peking Union Medical College Hospital (CAMS), Beijing, Beijing Municipality, China

Polyphenol-rich sugarcane extract (PRSE) is known to contain bioactive compounds with potential hypoglycemic properties, but its direct interaction with α-glucosidase remains unexplored. This study aimed to clarify the inhibitory mechanism of PRSE on α-glucosidase and to identify the key active compounds responsible for this effect. To achieve this, the inhibitory mechanism of PRSE on αglucosidase was investigated using enzyme kinetics, bioactive compounds with α-glucosidasebinding affinity through biolayer interferometry-mass spectrometry (BLI-MS), and their binding mechanisms were further explored via molecular docking analysis. Results revealed that PRSE inhibits α-glucosidase through a mixed-type mechanism. A total of 29 compounds were identified in the PRSE dissociation solution, including 4 coumarins and their derivatives, 9 phenolic acids and their derivatives, and 16 flavonoids. Representative compounds included coumarin, kaempferol, apigenin 7-o-neohesperidoside, and vicenin 3. Notably, apigenin 7-o-neohesperidoside and vicenin 3 were identified for the first time as potential α-glucosidase inhibitors. These compounds interact with key residues of α-glucosidase, such as Asp and Glu, via multiple molecular interactions, including hydrogen bonding, π-anion interactions, and hydrophobic forces. These findings suggest that PRSE could serve as a promising natural source of α-glucosidase inhibitors and demonstrate the efficiency of BLI-MS as a rapid and effective screening tool for identifying target bioactive compounds in plant extracts. Furthermore, this study supports the potential application of PRSE in functional foods for postprandial glycemic control and type 2 diabetes prevention.