Production, Characterization and Immunomodulatory Activity of an Extracellular Polysaccharide from Rhodotorula mucilaginosa YL-1 Isolated from Sea Salt Field
Abstract
:1. Introduction
2. Results
2.1. Identification of Strain and Phylogenetic Tree
2.2. Polysaccharide Production Optimization
2.3. Purification and Structural Characterization of YL-1 Polysaccharide
2.3.1. Determination of Homogeneity and Molecular Weight of Polysaccharide
2.3.2. Monosaccharide Composition Analysis
2.3.3. Sulfate Content Determination
2.3.4. FT-IR Spectrometric Analysis
2.3.5. Methylation and GC-MS Analysis
2.4. Bioactivity Test of Polysaccharide on RAW264.7 Cell Lines
2.4.1. MTT Test of the Cell Viability
2.4.2. NO Release Test
2.4.3. Neutral Red Uptake Test
2.4.4. Cytokine Concentration Test
2.4.5. Q-PCR Test Result of Cytokines
3. Discussion
4. Materials and Methods
4.1. Materials
4.2. Reagents
4.3. Production Optimization of Polysaccharides
4.4. Extraction and Purification of Extracellular Polysaccharides
4.5. Structure Characterization of Purified Polysaccharide
4.5.1. Determination of Homogeneity and Molecular Weight
4.5.2. Monosaccharide Composition Analysis
4.5.3. Sulfate Content Determination
4.5.4. FT-IR Spectrometric Analysis
4.5.5. Methylation and GC-MS Analysis
4.6. Immunomodulatory Activity of Purified Polysaccharide
4.6.1. Cell Viability
4.6.2. Immunomodulatory Activity of Cytokines
5. Conclusions
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
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Retention Time (min) | Monosaccharide Composition | Relative Molar Ratio |
---|---|---|
20.709 | rhamnose | 0.000 |
20.937 | fucose | 0.019 |
21.42 | arabinose | 0.000 |
22.075 | xylose | 0.000 |
31.52 | mannose | 0.276 |
31.763 | glucose | 0.176 |
32.562 | galactose | 0.529 |
RT | Methylated Sugar | Mass Fragments (m/z) | Molar Ratio | Type of Linkage |
---|---|---|---|---|
12 | 2,3,4-Me3-Fucp | 43, 59, 72, 89, 101, 115, 117, 131, 175 | 1.17 | Fuc-(1→ |
16.26 | 2,3,4,6-Me4-Glcp | 43, 71, 87, 101, 117, 129, 145, 161, 205 | 15 | Glc-(1→ |
16.556 | 2,3,4,6-Me4-Manp | 43, 71, 87, 101, 117, 129, 145, 161, 205 | 5.51 | Man-(1→ |
22.216 | 2,4,6-Me3-Galp | 43, 87, 99, 101, 117, 129, 161, 173, 233 | 52.8 | →3)-Gal-(1→ |
22.608 | 2,3,4-Me3-Manp | 43, 71, 87, 99, 101, 117, 129, 159, 161 | 3.84 | →6)-Man-(1→ |
25.074 | 4,6-Me2-Manp | 43, 85, 87, 99, 101, 127, 129, 161, 201, 216 | 5.36 | →2,3)-Man-(1→ |
29.548 | 2,4-Me2-Manp | 43, 87, 117, 129, 159, 189, 233 | 12.38 | →3,6)-Man-(1→ |
27.311 | 2,3-Me2-Glcp | 43, 71, 85, 87, 99, 101, 117, 127, 159, 161, 201 | 3.94 | →4,6)-Glc-(1→ |
Gene Name | Up Primers | Down Primers |
---|---|---|
TNF-α | 5-TGTCTACTGAACTTCGGGGTGAT-3 | 5-AACTGATGAGAGGGAGGCCAT-3 |
IL-1β | 5-AGTTGACGGACCCCAAAAG-3 | 5-AGCTGGATGCTCTCATCAGG-3 |
IL-6 | 5-GCTACCAAACTGGATATAATCAGGA-3 | 5-CCAGGTAGCTATGGTACTCCAGAA-3 |
IL-18 | 5-GCCATGTCAGAAGACTCTTGCGTC-3 | 5GTACAGTGAAGTCGGCCAAAGTTGTC-3 |
Actin | 5-GATTACTGCTCTGGCTCCTAGC-3 | 5-GACTCATCGTACTCCTGCTTGC-3 |
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Li, H.; Huang, L.; Zhang, Y.; Yan, Y. Production, Characterization and Immunomodulatory Activity of an Extracellular Polysaccharide from Rhodotorula mucilaginosa YL-1 Isolated from Sea Salt Field. Mar. Drugs 2020, 18, 595. https://doi.org/10.3390/md18120595
Li H, Huang L, Zhang Y, Yan Y. Production, Characterization and Immunomodulatory Activity of an Extracellular Polysaccharide from Rhodotorula mucilaginosa YL-1 Isolated from Sea Salt Field. Marine Drugs. 2020; 18(12):595. https://doi.org/10.3390/md18120595
Chicago/Turabian StyleLi, Haifeng, Lifeng Huang, Yunyi Zhang, and Yu Yan. 2020. "Production, Characterization and Immunomodulatory Activity of an Extracellular Polysaccharide from Rhodotorula mucilaginosa YL-1 Isolated from Sea Salt Field" Marine Drugs 18, no. 12: 595. https://doi.org/10.3390/md18120595