Scientia Agricultura Sinica ›› 2026, Vol. 59 ›› Issue (15): 3412-3423.doi: 10.3864/j.issn.0578-1752.2026.15.013

• FOOD SCIENCE AND ENGINEERING • Previous Articles     Next Articles

Flavonoid Composition and Gut Homeostasis-Modulatory Effects of Shatianyu (Citrus grandis L. Osbeck) Whole Fruit Fermented by Lactobacillus rhamnosus

MA XIAONI1,3(), WANG JIANMING1, ZHANG MINGWEI3, ZHANG LIN2, LUO GUANGYI3, ZHANG RUIFEN1,3, DENG MEI2()   

  1. 1 College of Food Science and Engineering, Tianjin University of Science & Technology, Tianjin 300457
    2 College of Food Science, South China Agricultural University, Guangdong Provincial Key Laboratory of Food Quality and Safety, Guangzhou 510642
    3 Sericultura & Agri-Food Research Institute, Guangdong Academy of Agricultural Sciences/Key Laboratory of Functional Foods, Ministry of Agriculture/Guangdong Key Laboratory of Agricultural Products Processing, Guangzhou 510610
  • Received:2025-12-29 Accepted:2026-04-24 Online:2026-08-01 Published:2026-08-03
  • Contact: DENG MEI

Abstract:

【Objective】This study aimed to elucidate the differences in flavonoid composition and gut homeostasis-regulating effects of Shatianyu (Citrus grandis L. Osbeck) whole fruit before and after debittering by Lactobacillus rhamnosus fermentation, thereby providing the theoretical foundation for the precise nutritional processing of Shatianyu.【Method】Shatianyu whole fruit powder (SWFP) and Lactobacillus rhamnosus-fermented Shatianyu whole fruit powder (F-SWFP) were used as test materials. High-performance liquid chromatography (HPLC) was adopted to analyze the differences in flavonoid composition and content between the above-mentioned fruit powder. Subsequently, SWFP and F-SWFP were subjected to in vitro fecal microbiota fermentation for 24 h, respectively. Gas chromatography (GC) and a lipopolysaccharide (LPS) enzyme-linked immunosorbent assay (ELISA) kit were employed to determine the content of short-chain fatty acids (SCFAs) and LPS in each fermentation system, respectively. 16S rRNA sequencing was adopted to determine the structure of fecal microbiota. The correlation between the relative abundance of fecal microbiota among SCFAs and LPS contents was revealed by Pearson correlation analysis.【Result】Eight monomeric flavonoids, namely neoeriocitrin, naringin, melitidin, cigranoside B, cigranoside A, cigranoside C, rhoifolin, and hesperidin, were detected in both SWFP and F-SWFP. Naringin, melitidin, and cigranoside B were the major flavonoid components, accounting for approximately 85% of the total content of detected flavonoids. Fermentation with Lactobacillus rhamnosus significantly reduced the total flavonoid content in SWFP, cigranoside B, neoeriocitrin, rhoifolin, hesperidin and melitidin were the main degraded flavonoids in F-SWFP, with the degradation rates ranging from 28.1% to 78.5%. After 24 h-fecal microbiota fermentation, both SWFP and F-SWFP significantly reduced the relative abundance ratio of Bacillota to Bacteroidota in fecal microbiota, with F-SWFP exhibiting a more pronounced effect. SWFP specifically induced the proliferation of Bifidobacterium and Mediterraneibacter, whereas F-SWFP not only specifically promoted the proliferation of Segatella and g_norank_f_Prevotellaceae, but also inhibited the growth of Collinsella, Blautia, Parabacteroides, and Dialister. 24 h-fecal microbiota fermentation of SWFP or F-SWFP all significantly increased acetic, propionic, and butyric acid content but reduced LPS content in the fermentation system. Notably, F-SWFP fermentation group showed higher SCFAs but lower LPS content than those of SWFP fermentation group, also, the butyric acid and LPS content in F-SWFP group was 1.2 and 0.7 times those of SWFP group, respectively. Correlation analysis revealed that the characteristic microbes in SWFP group were significantly positively correlated with SCFAs content, whereas those in F-SWFP group were not only positively correlated with SCFAs content, but also negatively correlated with LPS content.【Conclusion】Compared with SWFP, F-SWFP showed a lower total flavonoid content, but exhibited greater efficacy in promoting butyrate production while inhibiting LPS production by fecal microbiota. Given the key roles of butyrate and LPS in obesity-related metabolic diseases, F-SWFP serves as a functional ingredient for ameliorating obesity-related metabolic diseases and is suitable for use in the development of healthy foods.

Key words: Citrus grandis L. Osbeck, Lactobacillus rhamnosus fermentation, fecal microbiota fermentation, SCFAs, lipopolysaccharide (LPS)

Table 1

Composition and content of flavonoid components in SWFP and F-SWFP,respectively"

单体黄酮
Monomer flavonoid
沙田柚全果粉
SWFP
(mg·g-1 DW)
鼠李糖乳杆菌发酵沙田柚全果粉
F-SWFP (mg·g-1 DW)
新圣草次苷 Neoeriocitrin 1.97±0.14 0.86±0.01*
柚皮苷 Naringin 20.65±1.19 17.88±0.33*
麦利他汀Melitidin 13.58±1.61 9.77±0.19*
Cigranoside B 10.36±0.66 2.23±0.04**
Cigranoside A 1.91±0.10 1.49±0.03*
Cigranoside C 2.70±0.35 2.07±0.15
野漆树苷 Rhoifolin 0.71±0.03 0.33±0.01*
橙皮苷 Hesperidin 1.17±0.16 0.67±0.01*
总黄酮 Total flavone 53.05±3.40 35.29±0.59*

Fig. 1

Effects of SWFP and F-SWFP on the Alpha and Beta diversity of fecal microbiota Blank: Control group; SWFP: Shatianyu whole fruit powder fermentation group; F-SWFP: Lactobacillus rhamnosus-fermented SWFP fermentation group; Different lowercase letters indicate significant difference (P<0.05). The same as below"

Fig. 2

Effects of SWFP and F-SWFP on fecal microbial composition at phylum and family levels"

Fig. 3

Identification of the characteristic microbes in different fermentation groups by LEfSe analysis A: Multilevel species hierarchy of the LEfSe analysis; B: Characteristic microbial taxa among groups meeting an LDA significance threshold > 3"

Fig. 4

SCFAs contents in SWFP and F-SWFP fermentation groups"

Fig. 5

LPS content in SWFP and F-SWFP fermentation groups"

Fig. 6

Correlation analysis among the microbial genera with top 30 total relative abundance in each fermentation group and SCFAs, LPS contents in fermentation broth (23 genera with significant correlations are shown in the figure) A: Enrichment of genera significantly correlated with SCFAs and LPS content in SWFP and F-SWFP groups, respectively; ○ and ● indicate that the relative abundance of the genus in F-SWFP group is significantly lower or higher than those in SWFP group, respectively (P<0.05). Statistical analysis is performed using the Wilcoxon rank-sum test. B: Genera significantly correlated with SCFAs and LPS content in each fermentation group. Colors of squares represent the Pearson correlation (r-value). *, **, and *** indicate P<0.05, P<0.01 and P<0.001, respectively"

[1]
Li Q Y, Gao B, Siqin B, He Q, Zhang R, Meng X X, Zhang N H, Zhang N, Li M H. Gut microbiota: A novel regulator of cardiovascular disease and key factor in the therapeutic effects of flavonoids[J]. Frontiers in Pharmacology, 2021, 12: 651926.
[2]
Lin L, Zhang J Q. Role of intestinal microbiota and metabolites on gut homeostasis and human diseases[J]. BMC Immunology, 2017, 18(1): 2.
[3]
Gong L X, Wen T T, Wang J. Role of the microbiome in mediating health effects of dietary components[J]. Journal of Agricultural and Food Chemistry, 2020, 68(46)12820-12835.
[4]
Yang L, Gao Y C, Farag M A, Gong J P, Su Q L, Cao H, Zhang W, Zhao Y H, Wang H. Dietary flavonoids and gut microbiota interaction: A focus on animal and human studies to maximize their health benefits[J]. Food Frontiers, 2023, 4(4): 1794-1809.
[5]
Deng M, Jia X C, Dong L H, Liu L, Huang F, Chi J W, Ma Q, Zhao D, Zhang M W, Zhang R F. Structural elucidation of flavonoids from Shatianyu (Citrus grandis L. Osbeck) pulp and screening of key antioxidant components[J]. Food Chemistry, 2022, 366: 130605.
[6]
Deng M, Ye J M, Zhang R F, Zhang S, Dong L H, Su D X, Zhang M W, Huang F. Shatianyu (Citrus grandis L. Osbeck) whole fruit alleviated loperamide-induced constipation via enhancing gut microbiota-mediated intestinal serotonin secretion and mucosal barrier homeostasis[J]. Food & Function, 2024, 15(21): 10614-10627.
[7]
Singh S, Mishra S. Organoleptic and morphological analysis of Citrus peel powder[J]. Asian Food Science Journal, 2022: 1-11.
[8]
Gupta A K, Dhua S, Pratiksha, Kumar V, Naik B, Magwaza L S, Ncama K, Opara U L, McClements D J, Mishra P. Current and emerging applications in detection and removal of bitter compounds in Citrus fruit juice: A critical review[J]. Food Bioscience, 2023, 55: 102995.
[9]
叶佳敏, 张名位, 卢琦, 张瑞芬, 邓梅. 乳酸菌半固态发酵对沙田柚果粉苦味及活性成分的影响[J]. 中国农业科学, 2024, 57(13): 2662-2673. DOI: 10.3864/j.issn.0578-1752.2024.13.013.
Ye J M, Zhang M W, Lu Q, Zhang R F, Deng M. Effects of semi-solid fermentation with Lactobacillus on the bitterness and active components of Shatianyu (Citrus grandis L. Osbeck) fruit powder[J]. Scientia Agricultura Sinica, 2024, 57(13): 2662-2673. DOI: 10.3864/j.issn.0578-1752.2024.13.013.(in Chinese)
[10]
He J, Zhang P W, Shen L Y, Niu L L, Tan Y, Chen L, Zhao Y, Bai L, Hao X X, Li X W, Zhang S H, Zhu L. Short-chain fatty acids and their association with signalling pathways in inflammation, glucose and lipid metabolism[J]. International Journal of Molecular Sciences, 2020, 21(17): 6356.
[11]
Hine A M, Loke P. Intestinal macrophages in resolving inflammation[J]. The Journal of Immunology, 2019, 203(3): 593-599.
[12]
Sittipo P, Shim J W, Lee Y K. Microbial metabolites determine host health and the status of some diseases[J]. International Journal of Molecular Sciences, 2019, 20(21): 5296.
[13]
Gomes J M G, de Assis Costa J, de Cássia Gonçalves Alfenas R. Metabolic endotoxemia and diabetes mellitus: A systematic review[J]. Metabolism, 2017, 68: 133-144.
[14]
Wassenaar T M, Zimmermann K. Lipopolysaccharides in food, food supplements, and probiotics: Should we be worried?[J]. European Journal of Microbiology & Immunology, 2018, 8(3): 63-69.
[15]
Gonçalves P, Araújo J R, di Santo J P. A cross-talk between microbiota-derived short-chain fatty acids and the host mucosal immune system regulates intestinal homeostasis and inflammatory bowel disease[J]. Inflammatory Bowel Diseases, 2018, 24(3): 558-572.
[16]
Xiong R G, Zhou D D, Wu S X, Huang S Y, Saimaiti A, Yang Z J, Shang A, Zhao C N, Gan R Y, Li H B. Health benefits and side effects of short-chain fatty acids[J]. Foods, 2022, 11(18): 2863.
[17]
Gasaly N, Hermoso M A, Gotteland M. Butyrate and the fine-tuning of colonic homeostasis: Implication for inflammatory bowel diseases[J]. International Journal of Molecular Sciences, 2021, 22(6): 3061.
[18]
Salvi P S, Cowles R A. Butyrate and the intestinal epithelium: Modulation of proliferation and inflammation in homeostasis and disease[J]. Cells, 2021, 10(7): 1775.
[19]
Hu M Y, Zhang L, Ruan Z, Han P H, Yu Y J. The regulatory effects of Citrus peel powder on liver metabolites and gut flora in mice with non-alcoholic fatty liver disease (NAFLD)[J]. Foods, 2021, 10(12): 3022.
[20]
Tung Y C, Chang W T, Li S M, Wu J C, Badmeav V, Ho C T, Pan M H. Citrus peel extracts attenuated obesity and modulated gut microbiota in mice with high-fat diet-induced obesity[J]. Food & Function, 2018, 9(6): 3363-3373.
[21]
Ferreira-Lazarte A, Plaza-Vinuesa L, de las Rivas B, Villamiel M, Muñoz R, Moreno F J. Production of α-rhamnosidases from Lactobacillus plantarum WCFS1 and their role in deglycosylation of dietary flavonoids naringin and rutin[J]. International Journal of Biological Macromolecules, 2021, 193: 1093-1102.
[22]
Wang Z W, Tang H R, Liu G W, Gong H X, Li Y G, Chen Y L, Yang Y X. Compound probiotics producing cellulase could replace cellulase preparations during solid-state fermentation of millet bran[J]. Bioresource Technology, 2023, 385: 129457.
[23]
Paiva F F, Vanier N L, De Jesus Berrios J, Pan J, de Almeida Villanova F, Takeoka G, Elias M C. Physicochemical and nutritional properties of pigmented rice subjected to different degrees of milling[J]. Journal of Food Composition and Analysis, 2014, 35(1): 10-17.
[24]
王璐璐, 张名位, 叶佳敏, 张瑞芬, 邓梅. 沙田柚果肉可溶和不溶性膳食纤维对肠道菌群的影响[J]. 中国农业科学, 2024, 57(20): 4119-4129. DOI: 10.3864/j.issn.0578-1752.2024.20.016.
Wang L L, Zhang M W, Ye J M, Zhang R F, Deng M. Effects of soluble and insoluble dietary fiber from Shatianyu pulp on gut microbiota[J]. Scientia Agricultura Sinica, 2024, 57(20): 4119-4129. DOI: 10.3864/j.issn.0578-1752.2024.20.016.(in Chinese)
[25]
Cushnie T P T, Lamb A J. Antimicrobial activity of flavonoids[J]. International Journal of Antimicrobial Agents, 2005, 26(5): 343-356.
[26]
Gao P, Zheng M Y, Lu H Y, Lu S M. The progressive utilization of ponkan peel residue for regulating human gut microbiota through sequential extraction and modification of its dietary fibers[J]. Foods, 2023, 12(22): 4148.
[27]
Duda-Chodak A. The inhibitory effect of polyphenols on human gut microbiota[J]. Journal of Physiology and Pharmacology, 2012, 63(5): 497-503.
[28]
Ramos F M M, Ribeiro C B, Cesar T B, Milenkovic D, Cabral L, Noronha M F, Sivieri K. Lemon flavonoids nutraceutical (Eriomin®) attenuates prediabetes intestinal dysbiosis: A double-blind randomized controlled trial[J]. Food Science & Nutrition, 2023, 11(11): 7283-7295.
[29]
van den Broek L A, Hinz S W, Beldman G, Vincken J P, Voragen A G. Bifidobacterium carbohydrases-their role in breakdown and synthesis of (potential) prebiotics[J]. Molecular Nutrition & Food Research, 2008, 52(1): 146-163.
[30]
Xie Y X, Yang W J, Tang F, Chen X Q, Ren L C. Antibacterial activities of flavonoids: Structure-activity relationship and mechanism[J]. Current Medicinal Chemistry, 2014, 22(1): 132-149.
[31]
Araya-Cloutier C, Vincken J P, van de Schans M G M, Hageman J, Schaftenaar G, den Besten H M W, Gruppen H. QSAR-based molecular signatures of prenylated (iso)flavonoids underlying antimicrobial potency against and membrane-disruption in Gram positive and Gram negative bacteria[J]. Scientific Reports, 2018, 8: 9267.
[32]
Di Vincenzo F, Del Gaudio A, Petito V, Lopetuso L R, Scaldaferri F. Gut microbiota, intestinal permeability, and systemic inflammation: A narrative review[J]. Internal and Emergency Medicine, 2024, 19(2): 275-293.
[33]
Xiao X Y, Singh A, Giometto A, Brito I L. Segatella clades adopt distinct roles within a single individual’s gut[J]. npj Biofilms and Microbiomes, 2024, 10: 114.
[34]
Qu Y F, Li X Y, Xu F Y, Zhao S M, Wu X M, Wang Y Z, Xie J M. Kaempferol alleviates murine experimental colitis by restoring gut microbiota and inhibiting the LPS-TLR4-NF-κB axis[J]. Frontiers in Immunology, 2021, 12: 679897.
[35]
Zhan J, Liang Y R, Liu D H, Ma X R, Li P Z, Zhai W J, Zhou Z Q, Wang P. Pectin reduces environmental pollutant-induced obesity in mice through regulating gut microbiota: A case study of p, p′-DDE[J]. Environment International, 2019, 130: 104861.
[36]
Gomez-Arango L F, Barrett H L, Wilkinson S A, Callaway L K, McIntyre H D, Morrison M, Dekker Nitert M. Low dietary fiber intake increases Collinsella abundance in the gut microbiota of overweight and obese pregnant women[J]. Gut Microbes, 2018, 9(3): 189-201.
[37]
Strati F, Cavalieri D, Albanese D, De Felice C, Donati C, Hayek J, Jousson O, Leoncini S, Renzi D, Calabrò A, De Filippo C. New evidences on the altered gut microbiota in autism spectrum disorders[J]. Microbiome, 2017, 5(1): 24.
[38]
Mena-Vázquez N, Ruiz-Limón P, Moreno-Indias I, Manrique-Arija S, Lisbona-Montañez J M, Rioja J, Mucientes A, Martin-Núñez G M, Cano-García L, Tinahones F J, Fernández-Nebro A. Adiposity is associated with expansion of the genus Dialister in rheumatoid arthritis patients[J]. Biomedicine & Pharmacotherapy, 2023, 160: 114388.
[39]
Tito R Y, Cypers H, Joossens M, Varkas G, Van Praet L, Glorieus E, van den Bosch F, De Vos M, Raes J, Elewaut D. Brief report: Dialister as a microbial marker of disease activity in spondyloarthritis[J]. Arthritis & Rheumatology, 2017, 69(1): 114-121.
[1] WANG LuLu, ZHANG MingWei, YE JiaMin, ZHANG RuiFen, DENG Mei. Effects of Soluble and Insoluble Dietary Fiber from Shatianyu Pulp on Gut Microbiota [J]. Scientia Agricultura Sinica, 2024, 57(20): 4119-4129.
[2] YE JiaMin, ZHANG MingWei, LU Qi, ZHANG RuiFen, DENG Mei. Effects of Semi-Solid Fermentation with Lactobacillus on the Bitterness and Active Components of Shatianyu (Citrus grandis L. Osbeck) Fruit Powder [J]. Scientia Agricultura Sinica, 2024, 57(13): 2662-2673.
[3] LU YanQing, LIN YanJin, WANG XianDa, LU XinKun. A Transcriptome Analysis Identifies Candidate Genes Related to Fruit Cracking in Pomelo Fruits [J]. Scientia Agricultura Sinica, 2023, 56(20): 4087-4101.
[4] LU Qi,JIA XuChao,DENG Mei,ZHANG RuiFen,DONG LiHong,HUANG Fei,CHI JianWei,LIU Lei,ZHANG MingWei. Effects of Different Drying Methods on Bioactive Components of Shatianyou (Citrus grandis L. Osbeck) Pomace Powder [J]. Scientia Agricultura Sinica, 2022, 55(14): 2825-2836.
[5] YING ShiJia, DAI ZiChun, GUO JiaJia, SHI ZhenDan. Time Course Effect of Lipopolysaccharide on Toll-Like Receptors Expression in the Goose Follicular Stroma [J]. Scientia Agricultura Sinica, 2017, 50(6): 1147-1156.
Viewed
Full text


Abstract

Cited

  Shared   
  Discussed   
No Suggested Reading articles found!