Enzyme-aided-extraction, purification and characterization of anti-inflammatory compounds from seaweed /
Use of available anti-inflammatory drugs cause adverse side effects and even death, thus required alternative anti-inflammatory biocompounds extracted with environmental friendly method. Aqueous extracts, ethanolic precipitate and supernates of five Malaysian brown seaweeds; Sargassum duplicatum, Sa...
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| Main Author: | |
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| Format: | Thesis |
| Language: | English |
| Published: |
Kuala Lumpur :
Kulliyyah of Engineering, International Islamic University Malaysia,
2014
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| Subjects: | |
| Online Access: | Click here to view 1st 24 pages of the thesis. Members can view fulltext at the specified PCs in the library. |
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| Summary: | Use of available anti-inflammatory drugs cause adverse side effects and even death, thus required alternative anti-inflammatory biocompounds extracted with environmental friendly method. Aqueous extracts, ethanolic precipitate and supernates of five Malaysian brown seaweeds; Sargassum duplicatum, Sargassum binderi, Sargassum fulvellum, Padina australis and Turbinaria turbinata, were investigated for inhibition Nitric oxide (NO) production in lipopolysaccharide induced macrophage RAW 264.7 cell line. All extracts contain high fucose and inhibit NO secretion in a dose dependent manner, precipitate of T. turbinata was highest in percentage yield 20% and dosage of 200g/ml down-regulated >80% NO secretion. Among the supernates, that of T. turbinata, which contain highest fucose, gave highest inhibition of NO synthesis. Cytotoxicity assay revealed that some extracts were moderately toxic. Enzyme aided extraction was carried out on T. turbinata and release of ethanolic precipitate was enhanced by the use of cellulase compared to vicozymes and amyloglucosidase. Eight processing conditions were screened with Plackett-Burman design and 3 significant factors (hydrolysis time, enzyme concentration and extraction stage) were optimized using Faced Centered Central Composite Design in Response Surface Methods. The conditions, that gave 25.13 and 10.54 %, respectively for crude precipitated fraction and fucoidan yield, are extraction stages of 2, hydrolysis time of 19.5 h and enzyme concentration of 0.15 %. Increase in yield was due to substrate ruptured by cellulase according to micrograph images from Field Emission Scanning Electron Microscopy. Infrared spectra of sulfated polysaccharide confirmed presence of sulfated groups. Anion exchange chromatography separation gave three fractions: TtF1, TtF2 and TtF3. Yield and chemical composition of purified fractions varied considerably and monosaccharide composition showed high percentage of fucose (60.920.09 %) in TtF3 which suggests that the polysaccharide might be fucoidan with 41.710.29 % total sugar, 8.330.45 % uronic acid, 33.790.57 % sulfate ion. TtF1 and TtF2 lack anti-inflammatory potential while TtF3 inhibited NO secretion in LPS induced macrophage with IC50 value of 103.76 g/ml. Secretion of PGE2, TNF-α and IL-8 were also dose dependently inhibited by TtF3. Cytotoxicity assay confirmed that TtF3 was nontoxic. The sulfated polysaccharide fractions exhibited different molecular weight with 223.5, 495.5 and 326.05 kDa, respectively for TtF1, TtF2 and TtF3. Proton NMR signals showed that the spectra of TtF2 and TtF3 possessed α-anomeric proton (5– 5.6 ppm), ring proton (3.4 – 4.4), O-acetyl group (2.2 ppm) and methyl proton (1 – 1.3 ppm) while that of TtF1 only showed presense of ring proton and methyl proton. Rheological analysis showed that the sulphated polysaccharide fractions exhibited Newtonian and/or weak pseudoplastic behaviour when data were fitted with power law. Increase in consistency value was highest in TtF2 compared to that of TtF1 while TtF3 was lowest. Thermal degradation pattern of TtF1 and TtF2 were similar compared to that of TtF3. Both chemical and physical properties have some relationships with the anti-inflammatory role of the sulfated polysaccharide extract. The findings confirmed the potential of brown seaweed of Malaysia origin as a viable source of anti-inflammatory compounds. |
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| Physical Description: | xx, 218 leaves : ill. ; 30cm. |
| Bibliography: | Includes bibliographical references (leaves 194-211). |