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. 2024 Mar 11;29(6):1238.
doi: 10.3390/molecules29061238.

Biphasic Fermentation of Trapa bispinosa Shells by Ganoderma sinense and Characterization of Its Polysaccharides and Alcoholic Extract and Analysis of Their Bioactivity

Xiaoyan Sun  1 , Qiuqi Lei  2 , Qinyi Chen  1 , Dandan Song  1 , Min Zhou  2 , Hongxun Wang  1 , Limei Wang  1
Affiliations

Biphasic Fermentation of Trapa bispinosa Shells by Ganoderma sinense and Characterization of Its Polysaccharides and Alcoholic Extract and Analysis of Their Bioactivity

Xiaoyan Sun et al. Molecules. .

Abstract

Background: Trapa bispinosa shells (TBs) and its flesh (TBf) have been recognized for their medicinal properties, including antioxidant, antitumor, and immunomodulatory effects. Despite these benefits, TBs are often discarded as waste material, and their applications remain to be further explored.

Methods: In this study, we optimized the solid-state fermentation process of Ganoderma sinense (GS) with TBs using a response surface experiment methodology to obtain the fermented production with the highest water extract rate and DPPH free radical scavenging activity. We prepared and characterized pre-fermentation purified polysaccharides (P1) and post-fermentation purified polysaccharides (P2). Alcoholic extracts before (AE1) and after (AE2) fermentation were analyzed for active components such as polyphenols and flavonoids using UPLC-QTOF-MS/MS (ultra-performance liquid chromatography-quadrupole time-of-flight tandem mass spectrometry). Mouse macrophages (RAW 264.7) were employed to compare the immune-stimulating ability of polysaccharides and the antioxidant activity of AE1 and AE2.

Results: Optimal fermentation conditions comprised a duration of 2 days, a temperature of 14 °C, and a humidity of 77%. The peak water extract yield and DPPH free radical scavenging rate of the water extract from TBs fermented by GS were observed under these conditions. The enhanced activity may be attributed to changes in the polysaccharide structure and the components of the alcoholic extract. The P2 treatment group indicated more secretion of RAW 264.7 cells of NO, iNOS, IL-2, IL-10, and TNF-α than P1, which shows that the polysaccharides demonstrated increased immune-stimulating ability, with their effect linked to the NF-кB pathway. Moreover, the results of the AE2 treatment group indicated that secretion of RAW 264.7 cells of T-AOC and T-SOD increased and MDA decreased, which shows that the alcoholic extract demonstrated enhanced antioxidant activity, with its effect linked to the Nrf2/Keap1-ARE pathway.

Conclusions: Biphasic fermentation of Trapa bispinosa shells by Ganoderma sinense could change the composition and structure of the polysaccharides and the composition of the alcoholic extract, which could increase the products' immunomodulatory and antioxidant activity.

Keywords: Ganoderma sinense; Trapa bispinosa shell; UPLC-QTOF-MS/MS; antioxidant activity; immune activity; polysaccharides.

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Conflict of interest statement

The authors declare no conflicts of interest.

Figures

Figure 1
Figure 1
The single-factor experiment plots show the rate of DPPH radical scavenging and yield of water extract: (A) Fermentation humidity (%); (B) Fermentation temperature (°C); (C) Fermentation times (d).
Figure 2
Figure 2
Response surface plots showing effects of variables on the extraction yield of water extract: (A) Interaction of Temperature and Time; (B) Interaction of Humidity and Time; (C) Interaction of Humidity and Temperature.
Figure 3
Figure 3
The cytotoxicity models of RAW264.7 cells: (A) cytotoxicity of P1, P2 treatment; (B) cytotoxicity of AE1, AE2 treatment; (C) cytotoxicity and NO secretion of LPS treatment. The values in the figure are labeled with superscripts a-f in order from lowest to highest. Values with different superscripts are significantly different (p < 0.05). (D) cytotoxicity of H2O2 treatment.
Figure 4
Figure 4
The effect on inflammatory factors of P1 and P2: (A) secretion of NO; (B) secretion of iNOS; (C) secretion of TNF-α; (D) secretion of IL-2; (E) secretion of IL-10; (F)expression of genes related to NF-κB pathway. The values in the figure are labeled with superscripts a-c in order from lowest to highest. Values with different superscripts are significantly different (p < 0.05). Notes: 50 µg/mL P1, P2 compared with LPS treatment group and blank control group, Δ p < 0.05, ΔΔ p < 0.01; 100 µg/mL P1, P2 compared with LPS treatment group and blank control group, ** p < 0.01, *** p < 0.001; 150 µg/mL P1, P2 compared with LPS treatment group and blank control group, ## p < 0.01, ### p < 0.001.
Figure 5
Figure 5
The effects on antioxidant activity of AE1 and AE2: (A) secretion of MDA; (B) secretion of T-AOC; (C) secretion of T-SOD; (D) expression of genes related to Nrf2/Keap1-ARE pathway. The values in the figure are labeled with superscripts a-c in order from lowest to highest. Values with different superscripts are significantly different (p < 0.05). Note: 50 µg/mL L-Ascorbic acid (VC), AE1, AE2 treatment group compared with blank control group, Δ p < 0.05; 100 µg/mL L-Ascorbic acid (VC), AE1, AE2 treatment group compared with blank control group, * p < 0.05, ** p < 0.01, *** p < 0.001; 150 µg/mL L-Ascorbic acid (VC), AE1, AE2 treatment group compared with blank control group, ## p < 0.01, ### p < 0.001.
Figure 6
Figure 6
SEC−MALL−RI chromatograms of (P1) and (P2).
Figure 7
Figure 7
Morphological characteristics by scanning electron microscope of polysaccharides at different resolutions: (A), P1; (B), P2. The image scales are 1 μm, 500 nm and 200 nm.
Figure 8
Figure 8
Infrared detection spectrum of P1 and P2.
Figure 9
Figure 9
Distribution map of triple helix structure of P1 and P2.
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References

    1. Adkar P., Dongare A., Ambavade S., Bhaskar V. Trapa bispinosa Roxb.: A review on nutritional and pharmacological aspects. Adv. Pharmacol. Pharm. Sci. 2014;2014:959830. - PMC - PubMed
    1. Iwaoka Y., Suzuki S., Kato N., Hayakawa C., Kawabe S., Ganeko N., Uemura T., Ito H. Characterization and identification of bioactive polyphenols in the Trapa bispinosa Roxb. pericarp extract. Molecules. 2021;26:5802. doi: 10.3390/molecules26195802. - DOI - PMC - PubMed
    1. Wang L., Yin D., Fan Y., Min T., Yi Y., Wang H. Molecular mechanism of the anti-gastric cancer activity of 1, 2, 3, 6-tetra-O-galloyl-β-D-glucose isolated from Trapa bispinosa Roxb. shell in vitro. PLoS ONE. 2022;17:e0269013. doi: 10.1371/journal.pone.0269013. - DOI - PMC - PubMed
    1. Takeshita S., Ishioka Y., Yagi M., Uemura T., Yamada M., Yonei Y. The effects of water chestnut (Trapa bispinosa Roxb.) on the inhibition of glycometabolism and the improvement in postprandial blood glucose levels in humans. Glycative Stress Res. 2016;3:24–132.
    1. Tania Wahed T.W., Islam M.F., Ratna Sarkar R.S., Sonia Wahed W.S. Phytochemical, antioxidant and cytotoxicity study of the methanolic extracts of leaves of Trapa bispinosa Roxb. Int. J. Pharm. Sci. Res. 2014;10:4400–4405.