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Structural elucidation of mulberry leaf oligosaccharide and its selective promotion of gut microbiota to alleviate type 2 diabetes mellitus

Tenggen Hua,b,c, Yuanshan Yua,b, Jijun Wua, Yujuan Xua, Gengsheng Xiaod, Kejing Ana, Erna Lia, Sentai Liaoa, Yuxiao Zoua()
Sericultural & Agri-Food Research Institute, Guangdong Academy of Agricultural Sciences/Key Laboratory of Functional Foods, Ministry of Agriculture and Rural Affairs/Guangdong Key Laboratory of Agricultural Products Processing, Guangzhou 510610, China
Huagongliya (Foshan) Technology Industry Co., Ltd., Foshan 528313, China
Heyuan Branch, Guangdong Laboratory for Lingnan Modern Agriculture, Heyuan 517001, China
Zhongkai University of Agriculture and Engineering, Guangzhou 510225, China

Peer review under responsibility of Tsinghua University Press.

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Highlights

• A non-digestible oligosaccharide (MLO 2-1) was obtained.

• The proposed sugar chain structure of MLO 2-1 was determined.

• MLO 2-1 could tolerate simulated saliva, gastric and intestinal digestion.

• MLO 2-1 displayed the regulatory activity on the gut microbiota, especially Lactobacillus murinus.

• MLO 2-1 showed hypoglycemic activity by selectively accelerating the proliferation of L. murinus.

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Abstract

Two oligosaccharide fractions (MLO 2-1 and 2-2) were purified from enzymatic hydrolysate of mulberry leaf polysaccharide. The results of simulated digestion showed that MLO 2-2 was a digestible oligosaccharide, which could be degraded by human digestive juice; while MLO 2-1 possessed the non-digestible property in the upper gastrointestinal tract and performed the function by regulating the gut microbiota. Hence, MLO 2-1 was selected for the further analysis. The structure of MLO 2-1 was elucidated as follow: α-T-Glcp-(1→3)-β-Glcp-(1→5)-α-Araf-(1→5)-α-Araf-1→5)-α-Araf-(1→3)-α-(2-OAc)-Glcp-1. The in vitro fecal fermentation results showed that MLO 2-1 could modulate the composition of gut microbiota. Meanwhile, MLO 2-1 was effectively metabolized by fecal bacteria to produce lactate and short chain fatty acids, especially acetate and butyrate. The specific metabolic pathways of MLO 2-1 by gut microbiota were further illuminated. Gut microbiota analysis revealed that MLO 2-1 selectively promoted the growth of Ligilactobacillus murinus, a commensal bacterium presented a reduced level in T2DM mice. Animal experiments indicated that MLO 2-1 and L. murinus exhibited hypoglycemic activities. These results demonstrated that MLO 2-1 might alleviate T2DM by selectively accelerating the proliferation of L. murinus.

Keywords

Mulberry leaf oligosaccharideCharacterizationGut microbiotaLactobacillus murinusHypoglycemic activity

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Food Science and Human Wellness
Volume 13 Issue 4,
July 2024
Pages 2161-2173
DOI: 10.26599/FSHW.2022.9250180
Cite this article:
Hu T, Yu Y, Wu J, et al. Structural elucidation of mulberry leaf oligosaccharide and its selective promotion of gut microbiota to alleviate type 2 diabetes mellitus. Food Science and Human Wellness, 2024, 13(4): 2161-2173. https://doi.org/10.26599/FSHW.2022.9250180
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The methylation analysis results of MLO 2-1.

PMAARetention time (min)Molar ratioLinkage
1, 5-Di-O-acetyl-2, 3, 4, 6-tetra-O-methyl glucitol9.6081.00→1)-Glcp
1, 4, 5-Tri-O-acetyl-2, 3-di-O-methyl arabinitol11.3122.89→5)-Araf-(1→
1, 3, 5-Tri-O-acetyl-2, 4, 6-tri-O-methyl glucitol12.9562.13→3)-Glcp-(1→

The H and C chemical shift of MLO 2-1.

ResidueChemical shifts δ
123456
A (→3-β-Glcp-1→)H4.423.293.623.513.423.72
C103.3674.6874.5071.5876.9562.97
B (→3-α-Glcp-1→)H5.383.503.733.403.783.80
C93.1872.4172.2870.3172.8563.10
C (→5-α-Araf-1→)H5.394.043.914.143.76
C104.2680.1676.0580.6366.98
D (α-T-Glcp-1→)H5.333.513.713.423.823.83
C93.2672.7774.6870.0272.3760.86

Inter-residue cross peaks observed in HMBC spectrum of MLO 1-2.

ResidueSugar linkageH1/C1 δConnectivities
δH/δCδH/δCResidueAtom
Note: a Not observed.
A→3-β-Glcp-1→4.4266.98CC5
B→3-α-(2-OAc)-Glcp-1→5.38-a
OAc171.353.50BH2
C→5-α-Araf-1→5.3972.28BC3
Dα-T-Glcp-1→5.3374.50AC3