Production of Active Poly- and Oligosaccharidic Fractions from Ulva sp. by Combining Enzyme-Assisted Extraction (EAE) and Depolymerization
Abstract
:1. Introduction
2. Results
2.1. Extraction and Fractionation of Ulvans
2.1.1. Yields of Extraction and Fractionation
2.1.2. Biochemical Characterization of Ulvans
2.2. Oligosaccharide Production by Ulvans Depolymerization
2.2.1. Monitoring of Depolymerization by High Performance Size Exclusion Chromatography (HPSEC)
2.2.2. Yields of Oligosaccharide Production
2.2.3. Biochemical Characterization of Oligosaccharides
2.2.4. Matrix-Assisted Laser Desorption Ionization-Time of Flight (MALDI-TOF) Mass Spectrometry
2.3. Biological Activities
2.3.1. Lipoxygenase Inhibition Assay by Crude Ulvans and HMWP and LMWP Fractions from Ulva sp.
2.3.2. Modulation of Fibroblast Proliferation by Crude Ulvans and HMWP and LMWP Fractions from Ulva sp.
3. Discussion
3.1. Extraction and Fractionation of Ulvans
3.1.1. Ulvan Production
3.1.2. Biochemical Characterization of Ulvans
3.2. Depolymerization of Ulvans for Oligosaccharide Production
3.2.1. Oligosaccharide Production
3.2.2. Biochemical Characterization of Oligosaccharides
3.3. Biological Activities
3.3.1. Anti-Inflammatory Activity by Lipoxygenase Inhibition
3.3.2. Effects of the Ulvans Extracts and Fractions on Fibroblast Proliferation and Viability
4. Materials and Methods
4.1. Extraction and Fractionation Procedure (Figure 6)
4.1.1. Enzyme-Assisted Extraction (EAE) Followed by Polysaccharide Precipitation
4.1.2. Fractionation of Crude Ulvans
4.2. Production of Oligosaccharides by Ulvan Depolymerization
4.2.1. Hydrogen Peroxide Depolymerization
4.2.2. Depolymerization by Ion-Exchange Resin
4.3. Polysaccharide Molecular Weight (MW) Distribution by HPSEC Analysis
4.4. Biochemical Composition Analysis
4.5. Monosaccharide Composition Determined by High Performance Anion Exchange Chromatography
4.6. Matrix-Assisted Laser Desorption Ionisation-Time of Flight (MALDI-TOF) Mass Spectrometry
4.7. Biological Activities
4.7.1. Lipoxygenase Inhibition Assay
4.7.2. Cell Culture
4.7.3. WST-1 Assay
4.8. Statistical Analysis
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
Appendix A
References
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Mineral Matter | Carbohydrates | Uronic Acids | Sulfate Groups | Proteins | |
---|---|---|---|---|---|
Crude Ulvans | |||||
U | 35.2 ± 0.2 a | 23.5 ± 0.5 c | 18.5 ± 0.6 c | 29.9 ± 0.5 c | 10.5 ± 0.4 b,c |
HMWPs | |||||
PP-U | 26.8 ± 1.0 b | 30.7 ± 0.2 b | 23.7 ± 0.7 b | 41.9 ± 0.5 b | 11.2 ± 0.2 b,c |
DS-U | 13.4 ± 0.8 c | 37.4 ± 0.2 a | 37.0 ± 0.7 a | 49.1 ± 1.0 a | 12.8 ± 0.2 a,b |
AS-DS-U | - | 35.1 ± 0.4 a,b | 36.0 ± 0.8 a | 49.4 ± 0.5 a | 8.9 ± 0.5 c |
AS-DP-U | - | 31.5 ± 0.6 b | 13.3 ± 0.4 c | 23.9 ± 0.5 d | 16.4 ± 0.2 a |
Rhamnose | Galactose | Glucose | Xylose | Glucuronic Acid | |
---|---|---|---|---|---|
Crude Ulvans | |||||
U | 50.4 ± 6.0 a | 1.6 ± 0.1 b | 14.0 ± 1.2 a | 3.2 ± 0.2 a,b | 10.6 ± 0.9 a |
HMWPs | |||||
PP-U | 42.4 ± 0.3 a | 1.3 ± 0.1 b | 5.7 ± 0.7 b,c | 2.7 ± 0.2 b | 11.6 ± 1.9 a |
DS-U | 50.2 ± 2.2 a | 1.6 ± 0.0 b | 3.3 ± 0.2 c | 2.8 ± 0.2 a,b | 11.9 ± 0.3 a |
AS-DS-U | 59.0 a | 1.3 b | 4.0 b,c | 3.0 a,b | 11.2 a |
AS-DP-U | 29.1 a | 6.2 a | 9.0 b | 4.3 a | 3.4 b |
Mn (kDa) | Mw (kDa) | I | |
---|---|---|---|
PP-U | 1651 | 2290 | 1.4 |
DEP-H1 PP-U | 1480 | 2154 | 1.5 |
DEP-H3 PP-U | 1343 | 1943 | 1.5 |
DEP-H7 PP-U | 128 | 510 | 4.0 |
DEP-HD PP-U | 7.0 | 8.7 | 1.2 |
Carbohydrates | Uronic Acids | Sulfate Groups | Proteins | |
---|---|---|---|---|
DEP-AD PP-U | 30.4 ± 0.2 a | 30.8 ± 1.0 a | nd | 12.8 ± 1.0 a |
DEP-HD PP-U | 24.4 ± 0.4 b | 21.6 ± 0.4 b | 6.6 ± 0.1 | 16.8 ± 0.4 a |
Rhamnose | Galactose | Glucose | Xylose | Glucuronic Acid | |
---|---|---|---|---|---|
DEP-AD PP-U | 55.4 ± 2.3 a | 1.7 ± 0.1 a | 7.0 ± 0.4 a | 2.6 ± 0.2 a | 11.0 ± 0.7 a |
DEP-HD PP-U | 44.9 a | 1.0 b | 4.9 a | 2.1 a | 7.5 b |
Fractions | Temperature (°C) | Time (h) | Dialysis (500–1000 Da) |
---|---|---|---|
DEP-H1 PP-U | 35 | 1 | No |
DEP-H3 PP-U | Step 1: 35 Step 2: 50 | 1 2 | No |
DEP-H7 PP-U | Step 1: 35 Step 2: 50 | 1 6 | No |
DEP-HD PP-U | 50 | 24 | Yes |
Chromatographic Columns | |
---|---|
PP-U | G6000PWXL G4000PWXL G3000PWXL |
DEP-H1 PP-U | |
DEP-H3 PP-U | |
DEP-H7 PP-U | |
DEP-HD PP-U | G3000PWXL |
DEP-AD PP-U |
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Fournière, M.; Latire, T.; Lang, M.; Terme, N.; Bourgougnon, N.; Bedoux, G. Production of Active Poly- and Oligosaccharidic Fractions from Ulva sp. by Combining Enzyme-Assisted Extraction (EAE) and Depolymerization. Metabolites 2019, 9, 182. https://doi.org/10.3390/metabo9090182
Fournière M, Latire T, Lang M, Terme N, Bourgougnon N, Bedoux G. Production of Active Poly- and Oligosaccharidic Fractions from Ulva sp. by Combining Enzyme-Assisted Extraction (EAE) and Depolymerization. Metabolites. 2019; 9(9):182. https://doi.org/10.3390/metabo9090182
Chicago/Turabian StyleFournière, Mathilde, Thomas Latire, Marie Lang, Nolwenn Terme, Nathalie Bourgougnon, and Gilles Bedoux. 2019. "Production of Active Poly- and Oligosaccharidic Fractions from Ulva sp. by Combining Enzyme-Assisted Extraction (EAE) and Depolymerization" Metabolites 9, no. 9: 182. https://doi.org/10.3390/metabo9090182