Study on the structural characteristics, in vitro hypoglycemic activity, and regulatory mechanisms of Alpinia oxyphylla polysaccharide in modulating human gut microbiota
Main Article Content
Keywords
Alpinia oxyphylla, intestinal microbiota, polysaccharide, structural analysis
Abstract
This study aimed to systematically compare the structural characteristics, in vitro hypoglycemic activity, and regulatory effects on gut microbiota of Alpinia oxyphylla polysaccharides (AOFP) obtained by three extraction techniques, high-pressure and high-temperature (HH), acid-assisted high-pressure and high-temperature (AHP), and Aspergillus niger fermentation (ANF), to elucidate their structure–activity relationship and gut microbiota regulation mechanisms. By comparing the three types of AOFP (AOFP-HH, AOFP-AHP, AOFP-ANF), we revealed intrinsic associations among structure, activity, and microbiota regulation. The HH method promoted the ordered arrangement of β-1,3-glucan chains, forming a semicrystalline state and folded lamellar structures, thereby exhibiting the strongest hypoglycemic activity (α-amylase inhibition rate: 97.08 %; α-glucosidase inhibition rate: 59.43 %). The key mechanism lies in the synergy between the integrity of β-1,3-glycosidic bonds and the galacturonic acid content (11.74 %); together, they enhance activity through spatial hindrance effects and hydrogen bond networks. On the other hand, the AHP method, due to acid-induced cleavage of glycosidic bonds, generated amorphous low-molecular-weight structures (1.03 kDa), leading to a marked reduction in activity. The ANF method produced medium-molecular-weight fragments (1.14 kDa) via enzymatic hydrolysis. In terms of gut microbiota regulation, all AOFPs significantly promoted early butyrate production; notably, the HH group maintained a butyrate concentration of 0.076 mmol/L at 48 h. Each polysaccharide sample specifically increased the Firmicutes and stimulated the proliferation of Ligilactobacillus, thereby improving intestinal barrier function. Interestingly, the ANF group uniquely upregulated the abundance of Fusobacteriota, suggesting potential immunomodulatory capacity. Further analysis indicated that different processing techniques modulate selective microbial metabolic pathways through conformational differences. The AOFP-HH prepared by the HH method combines high hypoglycemic activity with structural stability, making it an ideal functional food additive. Meanwhile, the distinctive microbiota interaction mechanism exhibited by the ANF method offers novel insights for the development of personalized probiotics. In conclusion, our findings lay a solid theoretical foundation for the precise design of A. oxyphylla polysaccharides and their application in the prevention and treatment of -metabolic diseases.
References
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Chi, Y., Li, Y., Zhang, G., Gao, Y., Ye, H., Gao, J., et al. 2018. Effect of extraction techniques on properties of polysaccharides from Enteromorpha prolifera and their applicability in iron chelation. Carbohydrate Polymers. 181: 616–623. 10.1016/j.carbpol.2017.11.104
El-Gendi, H., Abu-Serie, M.M., Kamoun, E.A., Saleh, A.k., El-Fakharany, E.M. 2023. Statistical optimization and characterization of fucose-rich polysaccharides extracted from pumpkin (Cucurbita maxima) along with antioxidant and antiviral activities. International Journal of Biological Macromolecules. 232: 123372. 10.1016/j.ijbiomac.2023.123372
Ge, C.-C., Li, X.-Y., Qiao, W.-H., Cui, C., Wang, J., Gongpan, P., et al. 2024. BACE1 inhibitors from the fruits of Alpinia oxyphylla have efficacy to treat T2DM-related cognitive disorder. Fitoterapia. 178: 106157. 10.1016/j.fitote.2024.106157
Huang, K., Lin, M., Hsu, Y., Lu, I.H., Pan, I.H., Yang, J.L., et al. 2019. Alpinia oxyphylla fruit extract ameliorates experimental autoimmune encephalomyelitis through the regulation of Th1/Th17 Cells. Evidence-Based Complementary and Alternative Medicine. 2019: 6797030. 10.1155/2019/6797030
Jiang, L., Wang, W., Wen, P., Shen, M., Li, H., Ren, Y., et al. 2020. Two water-soluble polysaccharides from mung bean skin: Physicochemical characterization, antioxidant and antibacterial activities. Food Hydrocolloids. 100: 105412. 10.1016/j.foodhyd.2019.105412
Kac̆uráková, M., Capek, P., Sasinková, V., Wellner, N., Ebringerová, A. 2000. FT-IR study of plant cell wall model compounds: Pectic polysaccharides and hemicelluloses. Carbohydrate Polymers. 43(2): 195–203. 10.1016/S0144-8617(00)00151-X
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Li, H., Li, C., Xu, Y., Cao, H., Wang, X., He, J. 2025. Ultrasonic-assisted alkali extraction of quinoa polysaccharides: Yield and structural characterization. Journal of Cereal Science. 122: 104108. 10.1016/j.jcs.2025.104108
Li, X., and Wang, L., 2016. Effect of extraction method on structure and antioxidant activity of Hohenbuehelia serotine polysaccharides. International Journal of Biological Macromolecules. 83: 270–276. 10.1016/j.ijbiomac.2015.11.060
Li, Y., Zhou, X., and Fang, Z., 2024. Fang Zhaohui’s clinical experience in treating diabetic nephropathy with Yizhiren (Alpinia oxyphylla seeds). Clinical Journal of Traditional Chinese Medicine. 36(7): 1256–1259. 10.16448/j.cjtcm.2024.0714
Liberato, M.V., Prates, E.T., Gonçalves, T.A., Bernardes, A., Vilela, N., Fattori, J., et al. 2021. Insights into the dual cleavage activity of the GH16 laminarinase enzyme class on β-1,3 and β-1,4 glycosidic bonds. Journal of Biological Chemistry. 296: 100385. 10.1016/j.jbc.2021.100385
Lin, Y., Wu, J., Xiong, B., Wang, L., Zeng, Z., Zheng, B., et al. 2025. Effect of digestive properties and gut microbiota structure of steam-exploded Laminaria japonica polysaccharide: In vitro dynamic human gastrointestinal systems. LWT. 225: 117866. 10.1016/j.lwt.2025.117866
Liu, C., Bai, J., Yang, S., Shi J., Qin Y., Wu D., et al. 2024. Optimization of acid extraction process for Hovenia dulcis polysaccharides and investigation of its physicochemical properties and-antioxidant activities. Food and Fermentation Industries. 50(9): 148–156. 10.13995/j.cnki.11-1802/ts.035439
Liu, Z., Wang, M., Li, J., Zhao, Z., Wang, Z., Hu, Y., et al. 2025. Sulfated polysaccharide from Hizikia fusiforme selectively utilized by Bacteroides caccae and regulates its metabolite profile during in vitro fermentation. Food Bioscience. 69: 106816. 10.1016/j.fbio.2025.106816
Pan, L., Tu, Y., Li, Y., Wei, Z., Liu, L., Zhang, A., et al. 2025. Physicochemical analysis and hypoglycemic potential of Tremella aurantialba polysaccharides. Food and Fermentation Industries. 1–13. 10.13995/j.cnki.11-1802/ts.041529
Park, C.L., Kim, J.H., Jeon, J.S., Lee, J. H., Zhang, K., Guo, S., et al. 2022. Protective effect of Alpinia oxyphylla fruit against tert--butyl hydroperoxide-induced toxicity in HepG2 cells via Nrf2 activation and free radical scavenging and its active molecules. Antioxidants. 11: 1032. 10.3390/antiox11051032
Qian, J.-Y., Chen, W., Zhang, W.-M., Zhang, H. 2009. Adulteration identification of some fungal polysaccharides with SEM, XRD, IR and optical rotation: A primary approach. Carbohydrate Polymers. 78(3): 620–625. 10.1016/j.carbpol.2009.05.025
Ruan, X., 2021. Study on bioactive components of Alpinia-oxyphylla from Guangxi and product development. Nanchang: Nanchang University. 10.27232/d.cnki.gnchu.2021.003672
Shang, H., Chen, S., Li, R., Zhou, H., Wu, H., Song, H. 2018. Influences of extraction methods on physicochemical characteristics and activities of Astragalus cicer L. polysaccharides. Process Biochemistry. 73: 220–227. 10.1016/j.procbio.2018.07.016
Shi, F., Yan, X., Cheong, K.-L., Liu, Y. 2018. Extraction, purification, and characterization of polysaccharides from marine algae Gracilaria lemaneiformis with anti-tumor activity. Process Biochemistry. 73: 197–203. 10.1016/j.procbio.2018.08.011
Shi, W., Zhong, J., Zhang, Q., Yan, C. 2020. Structural characterization and antineuroinflammatory activity of a novel heteropolysaccharide obtained from the fruits of Alpinia oxyphylla. Carbohydrate Polymers. 229: 115405. 10.1016/j.carbpol.2019.115405
Song, H., Zhang, Y., Huang, Q., Wang, F., Wang, L., Xiong, L., et al. 2025. Extraction optimization, purification, characterization, and hypolipidemic activities of polysaccharide from pumpkin. International Journal of Biological Macromolecules. 141907. 10.1016/j.ijbiomac.2025.141907
Song, J., Wu, Y., Ma, X., Feng, L., Wang, Z., Jiang, G., et al. 2020. Structural characterization and α-glycosidase inhibitory-activity of a novel polysaccharide fraction from Aconitum coreanum. Carbohydrate Polymers. 230: 115586. 10.1016/j.carbpol.2019.115586
Sun, Y., Zeng, H., Huang, J., Zhao, Y., Cai, J., Sun, S., et al. 2025. Isolation, purification, structural characterization and in vitro hypolipidemic activity of Wuzhuyu polysaccharides. Science and Technology of Food Industry. 1–18 [2025-03-17]. 10.13386/j.issn1002-0306.2024070437
Tang, Y., Wei, Z., He, X., Ling, D., Qin, M., Yi, P., et al. 2024. A comparison study on polysaccharides extracted from banana flower using different methods: Physicochemical characterization, and antioxidant and antihyperglycemic activities. International Journal of Biological Macromolecules. 264(1): 130459. 10.1016/j.ijbiomac.2024.130459
Wang, L., Liu, H.-M., and Qin, G.-Y., 2017. Structure characterization and antioxidant activity of polysaccharides from Chinese quince seed meal. Food Chemistry. 234: 314–322. 10.1016/j.foodchem.2017.05.002
Wang, M., Hu, J., You, F., Ma, F., Peng, Y., Zhu, S., et al. 2025. Structural characteristics, in vitro activities, and gut microbiota regulatory mechanism of Alpinia oxyphylla oligosaccharides. Science and Technology of Food Industry. 1–17 [2025-10-24]. 10.13386/j.issn1002-0306.2025060074
Wang, R., Chen, P., Jia, F., Tang, J., Ma, F. 2012. Optimization of polysaccharides from Panax japonicus C.A. Meyer by RSM and its anti-oxidant activity. International Journal of Biological Macromolecules. 50(2): 331–336. 10.1016/j.ijbiomac.2011.12.023
Wang, X., Yan, H., Xu, Y., Li, D., Bao, Y., Dai, Z. 2024. Stability during colonic digestion and modulatory effects on fecal microbiota of different modified lutein nanoliposomes. Science and Technology of Food Industry. 45(16): 376–383. 10.13386/j.issn1002-0306.2023090276
Xiang, H., Wen, W., Xu, P., Qiu, H., Chu, C., Shao, Q., et al. 2024. Inhibition mechanism of α-glucosidase by three geranylated compounds: Kinetic, spectroscopic and molecular docking. Process Biochemistry. 136: 237–244. 10.1016/j.procbio.2023.11.034
Yang, W., Su, L., Wang, L., Wu, J., Chen, S. 2022. Alpha-glucanotransferase from the glycoside hydrolase family synthesizes α(1–6)-linked products from starch: Features and synthesis pathways of the products. Trends in Food Science & Technology. 128: 160–172. 10.1016/j.tifs.2022.08.001
Yang, X., 2020. Structural characteristics and immunomodulatory activity of polysaccharides from Alpinia oxyphylla. Haikou: Hainan University. 10.27073/d.cnki.ghadu.2020.001252
Yang, X., Zhou, S., Li, H., An, J., Li, C., Zhou, R., et al. 2021. Structural characterization of Alpiniae oxyphyllaefructus polysaccharide 2 and its activation effects on RAW264.7 macrophages. International Immunopharmacology. 97: 107708. 10.1016/j.intimp.2021.107708
Yin, D., Sun, X., Li, N., Guo, Y., Tian, Y., Wang, L. 2021. Structural properties and antioxidant activity of polysaccharides extracted from Laminaria japonica using various methods. Process Biochemistry. 111(2): 201–209. 10.1016/j.-procbio.2021.10.019
Yin, Q., Xu, W., Wu, K., Meng, P., Cai, J., Lin, C. 2024. Effects of different extraction methods on physicochemical properties and bioactivities of polygonatum polysaccharides. Modern Food Science and Technology. 40(3): 142–152. 10.13982/j.mfst.1673-9078.2024.3.0112
Yusoff, M.H.M., and Shafie, M.H., 2024. A review of in vitro antioxidant and antidiabetic polysaccharides: Extraction methods, physicochemical and structure-activity relationships. International Journal of Biological Macromolecules. 282(4): 137143. 10.1016/j.ijbiomac.2024.137143
Zeng, D., and Zhu, S., 2018. Purification, characterization, antioxidant and anticancer activities of novel polysaccharides extracted from Bachu mushroom. International Journal of Biological Macromolecules. 107(A): 1086–1092. 10.1016/j.ijbiomac.2017.09.088
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Apr 1, 2026
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Wu, J., Li, J., Ma, F., Huang, S., Ye, X., Yang, M., Yang, H., Peng, Y., Zhu, S., & Wang, M. (2026). Study on the structural characteristics, in vitro hypoglycemic activity, and regulatory mechanisms of Alpinia oxyphylla polysaccharide in modulating human gut microbiota. Italian Journal of Food Science, 38(2), 71–93. https://doi.org/10.15586/ijfs.v38i2.3430
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How to Cite
Wu, J., Li, J., Ma, F., Huang, S., Ye, X., Yang, M., Yang, H., Peng, Y., Zhu, S., & Wang, M. (2026). Study on the structural characteristics, in vitro hypoglycemic activity, and regulatory mechanisms of Alpinia oxyphylla polysaccharide in modulating human gut microbiota. Italian Journal of Food Science, 38(2), 71–93. https://doi.org/10.15586/ijfs.v38i2.3430