R. Huang <huangriming
scau.edu.cn; X. Zhang <zhangxiaoyong
Institute for Environmental and Climate Research, Jinan University, Guangzhou 511443, China, University Joint Laboratory of Guangdong Province, Hong Kong and Macao Region on Marine Bioresource Conservation and Exploitation, College of Marine Sciences, South China Agricultural University, Guangzhou 510642, China, Guangdong Provincial Key Laboratory of Nutraceuticals and Functional Foods, College of Food Science, South China Agricultural University, Guangzhou 510642, China
Recent studies have found that many marine microbial polysaccharides exhibit distinct immune activity. However, there is a relative scarcity of research on the immunomodulatory activity of marine fungal exopolysaccharides. A novel water-soluble fungal exopolysaccharide ASP-1 was isolated from the fermentation broths of marine coral-associated fungus Aspergillus pseudoglaucus SCAU265, and purified by Diethylaminoethyl-Sepharose-52 (DEAE-52) Fast Flow and Sephadex G-75. Structural analysis revealed that ASP-1 had an average molecular weight of 36.07 kDa and was mainly composed of (1→4)-linked α-D-glucopyranosyl residues, along with highly branched heteropolysaccharide regions containing 1,4,6-glucopyranosyl, 1,3,4-glucopyranosyl, 1,4,6-galactopyranosyl, T(terminal)-glucopyranosyl, T-mannopyranosyl, and T-galactopyranosyl residues. ASP-1 demonstrated significant effects on the proliferation, nitric oxide levels, and the secretion of cytokines TNF-α and IL-6 in macrophage RAW264.7 cells. Metabolomic analysis provided insights into the potential mechanisms of the immune regulation of ASP-1, suggesting its involvement in regulating immune function by modulating amino acid anabolism, particularly arginine synthesis and metabolism. These findings provide fundamental scientific data for further research on its accurate molecular mechanism of immunomodulatory activity.
Fig. 2f, table 2, ASP-1
13C NMR, NMR-2D, GC-MS, sugar analysis, FTIR, extraction, statistical analysis, methylation (Hakomori method), fermentation, H NMR, HPGPC, ethanol precipitation, UV-vis, metabolomics analysis, immunomodulatory activity
At a concentration of 10 to 1000 μg, ASP-1 did not exhibit cytotoxicity towards RAW 264.7 macrophages. ASP-1 exhibited In vitro remarkable immune-enhancing activity, as indicated by the significant elevation of NO, TNF- , and IL-6 levels in RAW264.7 macrophages.
NMR temperature was not specified; proposed the chemical structure of a water-soluble EPS (ASP-1). Published NMRC spectra was removed by CSDB staff due to multiple errors in the NMR assignment.
1H NMR data:
Linkage Residue H1 H2 H3 H4 H5 H6
4,4,4,6,4,4,3,4,2,4,6,4,3,4 bDManp
4,4,4,6,4,4,3,4,2,4,6,4,3,6,4 aDGalp 5.29 3.58 3.83 4.19 3.93 3.72
4,4,4,6,4,4,3,4,2,4,6,4,3,6,6,4,6,4 bDManp
4,4,4,6,4,4,3,4,2,4,6,4,3,6,6,4,6,6 aDGlcp
4,4,4,6,4,4,3,4,2,4,6,4,3,6,6,4,6 aDGlcp
4,4,4,6,4,4,3,4,2,4,6,4,3,6,6,4,4 aDGalp 5.29 3.58 3.83 4.19 3.93 3.72
4,4,4,6,4,4,3,4,2,4,6,4,3,6,6,4 aDGlcp
4,4,4,6,4,4,3,4,2,4,6,4,3,6,6,6 aDGlcp
4,4,4,6,4,4,3,4,2,4,6,4,3,6,6 aDGlcp
4,4,4,6,4,4,3,4,2,4,6,4,3,6 aDGlcp
4,4,4,6,4,4,3,4,2,4,6,4,3 aDGalp
4,4,4,6,4,4,3,4,2,4,6,4 aDGlcp
4,4,4,6,4,4,3,4,2,4,6,3,4 bDManp
4,4,4,6,4,4,3,4,2,4,6,3,6,4 aDGalp 5.29 3.58 3.83 4.19 3.93 3.72
4,4,4,6,4,4,3,4,2,4,6,3,6,6,4,6,4 bDManp
4,4,4,6,4,4,3,4,2,4,6,3,6,6,4,6,6 aDGlcp
4,4,4,6,4,4,3,4,2,4,6,3,6,6,4,6 aDGlcp
4,4,4,6,4,4,3,4,2,4,6,3,6,6,4,4 aDGalp 5.29 3.58 3.83 4.19 3.93 3.72
4,4,4,6,4,4,3,4,2,4,6,3,6,6,4 aDGlcp
4,4,4,6,4,4,3,4,2,4,6,3,6,6,6 aDGlcp
4,4,4,6,4,4,3,4,2,4,6,3,6,6 aDGlcp
4,4,4,6,4,4,3,4,2,4,6,3,6 aDGlcp
4,4,4,6,4,4,3,4,2,4,6,3 aDGalp
4,4,4,6,4,4,3,4,2,4,6 aDGlcp 4.91 3.51 3.87 3.51 3.83 3.88
4,4,4,6,4,4,3,4,2,4 bDGlcp 4.81 3.50 4.61 3.51 3.94 3.60
4,4,4,6,4,4,3,4,2,6 bDManp 5.27 3.55 3.64 3.87 4.08 3.46
4,4,4,6,4,4,3,4,2 aDGalp 5.28 3.54 3.81 3.57 3.74 3.60
4,4,4,6,4,4,3,4 aDManp 5.27 3.56 3.92 3.79 3.64 3.75
4,4,4,6,4,4,3,6 bDManp 5.27 3.55 3.64 3.87 4.08 3.46
4,4,4,6,4,4,3 aDGalp 5.28 3.54 3.81 3.57 3.74 3.60
4,4,4,6,4,4 aDGlcp 5.27 3.55 3.89 3.87 3.57 3.66-3.84
4,4,4,6,4 aDGlcp 5.33 3.55 3.73 3.58 3.93 3.69
4,4,4,6,4,4,3,4,2,4,6,4,4 aDGlcp 4.89 3.52 3.58 3.34 3.58 3.69
4,4,4,6 aDGlcp
4,4,4 aDGlcp 4.89 3.57 3.65 3.75 3.85 3.75
4,4,6 aDGlcp 4.89 3.52 3.58 3.34 3.58 3.69
4,4 aDGlcp 4.89 3.57 3.65 3.75 3.85 3.75
4,6 aDGlcp 4.89 3.52 3.58 3.34 3.58 3.69
4 aDGlcp 4.89 3.57 3.65 3.75 3.85 3.75
6 aDGlcp 4.89 3.52 3.58 3.34 3.58 3.69
aDGlcp
1H NMR data:
| Linkage | Residue | H1 | H2 | H3 | H4 | H5 | H6 |
|---|
| 4,4,4,6,4,4,3,4,2,4,6,4,3,4 | bDManp | |
| 4,4,4,6,4,4,3,4,2,4,6,4,3,6,4 | aDGalp | 5.29 | 3.58 | 3.83 | 4.19 | 3.93 | 3.72 |
| 4,4,4,6,4,4,3,4,2,4,6,4,3,6,6,4,6,4 | bDManp | |
| 4,4,4,6,4,4,3,4,2,4,6,4,3,6,6,4,6,6 | aDGlcp | |
| 4,4,4,6,4,4,3,4,2,4,6,4,3,6,6,4,6 | aDGlcp | |
| 4,4,4,6,4,4,3,4,2,4,6,4,3,6,6,4,4 | aDGalp | 5.29 | 3.58 | 3.83 | 4.19 | 3.93 | 3.72 |
| 4,4,4,6,4,4,3,4,2,4,6,4,3,6,6,4 | aDGlcp | |
| 4,4,4,6,4,4,3,4,2,4,6,4,3,6,6,6 | aDGlcp | |
| 4,4,4,6,4,4,3,4,2,4,6,4,3,6,6 | aDGlcp | |
| 4,4,4,6,4,4,3,4,2,4,6,4,3,6 | aDGlcp | |
| 4,4,4,6,4,4,3,4,2,4,6,4,3 | aDGalp | |
| 4,4,4,6,4,4,3,4,2,4,6,4 | aDGlcp | |
| 4,4,4,6,4,4,3,4,2,4,6,3,4 | bDManp | |
| 4,4,4,6,4,4,3,4,2,4,6,3,6,4 | aDGalp | 5.29 | 3.58 | 3.83 | 4.19 | 3.93 | 3.72 |
| 4,4,4,6,4,4,3,4,2,4,6,3,6,6,4,6,4 | bDManp | |
| 4,4,4,6,4,4,3,4,2,4,6,3,6,6,4,6,6 | aDGlcp | |
| 4,4,4,6,4,4,3,4,2,4,6,3,6,6,4,6 | aDGlcp | |
| 4,4,4,6,4,4,3,4,2,4,6,3,6,6,4,4 | aDGalp | 5.29 | 3.58 | 3.83 | 4.19 | 3.93 | 3.72 |
| 4,4,4,6,4,4,3,4,2,4,6,3,6,6,4 | aDGlcp | |
| 4,4,4,6,4,4,3,4,2,4,6,3,6,6,6 | aDGlcp | |
| 4,4,4,6,4,4,3,4,2,4,6,3,6,6 | aDGlcp | |
| 4,4,4,6,4,4,3,4,2,4,6,3,6 | aDGlcp | |
| 4,4,4,6,4,4,3,4,2,4,6,3 | aDGalp | |
| 4,4,4,6,4,4,3,4,2,4,6 | aDGlcp | 4.91 | 3.51 | 3.87 | 3.51 | 3.83 | 3.88 |
| 4,4,4,6,4,4,3,4,2,4 | bDGlcp | 4.81 | 3.50 | 4.61 | 3.51 | 3.94 | 3.60 |
| 4,4,4,6,4,4,3,4,2,6 | bDManp | 5.27 | 3.55 | 3.64 | 3.87 | 4.08 | 3.46 |
| 4,4,4,6,4,4,3,4,2 | aDGalp | 5.28 | 3.54 | 3.81 | 3.57 | 3.74 | 3.60 |
| 4,4,4,6,4,4,3,4 | aDManp | 5.27 | 3.56 | 3.92 | 3.79 | 3.64 | 3.75 |
| 4,4,4,6,4,4,3,6 | bDManp | 5.27 | 3.55 | 3.64 | 3.87 | 4.08 | 3.46 |
| 4,4,4,6,4,4,3 | aDGalp | 5.28 | 3.54 | 3.81 | 3.57 | 3.74 | 3.60 |
| 4,4,4,6,4,4 | aDGlcp | 5.27 | 3.55 | 3.89 | 3.87 | 3.57 | 3.66
3.84 |
| 4,4,4,6,4 | aDGlcp | 5.33 | 3.55 | 3.73 | 3.58 | 3.93 | 3.69 |
| 4,4,4,6,4,4,3,4,2,4,6,4,4 | aDGlcp | 4.89 | 3.52 | 3.58 | 3.34 | 3.58 | 3.69 |
| 4,4,4,6 | aDGlcp | |
| 4,4,4 | aDGlcp | 4.89 | 3.57 | 3.65 | 3.75 | 3.85 | 3.75 |
| 4,4,6 | aDGlcp | 4.89 | 3.52 | 3.58 | 3.34 | 3.58 | 3.69 |
| 4,4 | aDGlcp | 4.89 | 3.57 | 3.65 | 3.75 | 3.85 | 3.75 |
| 4,6 | aDGlcp | 4.89 | 3.52 | 3.58 | 3.34 | 3.58 | 3.69 |
| 4 | aDGlcp | 4.89 | 3.57 | 3.65 | 3.75 | 3.85 | 3.75 |
| 6 | aDGlcp | 4.89 | 3.52 | 3.58 | 3.34 | 3.58 | 3.69 |
| | aDGlcp | |
|
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scau.edu.cn; X. Zhang <zhangxiaoyong