碩士 / 國立臺灣海洋大學 / 食品科學系 / 107 / This aim of this study is to investigate the enzyme-assisted production conditions of fucooligosaccharides (FOS) from Laminaria japonica powder and the enzyme-assisted extraction conditions of fucoxanthin from L. japonica residue, as well as to evaluate their antioxidant activity and anti-inflammatory activity. Firstly, L. japonica fucoidan (LJ-FD) is gained from L. japonica treated by hot acid method, the yield is 18.09% (18.09 g per 100 g L. japonica powder). L. japonica fucoolisaccharides (LJ-FOS3k) is produced by hydrolyzing L. japonica powder with fucosidases which are induced by marine bacteria Pseudomonas vesicularis MA103, Aeromonas salmonicida MAEF108, and transgenic strain Lactococcus lactis NZ3900 rfucI-H. After 3% L. japonica powder is first hydrolyzed by commercial cellulase for 24 hr at pH 6.5 then fucosidases for 72 hr at pH 5.5, ultrafiltration system is used to collect < 3 kDa molecular, and the yield of LJ-FOS3k is 11.74% (11.74 g FOS3k per 100 g L. japonica powder). HPLC chromatogram results showed that there are 2,339 Da (DP 12) and 536 Da (DP 2) olisaccgarides in LJ-FOS3k, and the increased of hydrolysis time, the content of olisaccgarides has an upward trend. Secondly, using L. japonica residue (LJr) obtained from the above experiment to induce strains MA103 and MAEF108, the better total enzyme activities are 7.21 and 1.09 U/mL, respectively, which are produced by MA103 in 2 days and MAEF 108 in 3 days. Hydrolyzing 4% homogenous LJr with 30% (v/v) cude enzyme solution for 48 hr at 26oC, and then extract L. japonica fucoxanthin (LJ-FX) at room temperature (26oC) with 90% acetone, the concentration of FX in extract, quantified by absorbance value, is 6.061 g/mL. After initial separation and HPLC analysis, the yield of FX is 0.49‱ (0.49 mg LJ-FX per 100 g L. japonica powder). In antioxidant activity assays, FD (20 mg/mL), FOS3k (20 mg/mL), and FX (4 g/mL) have 30.42%, 43.95%, and 81.19% of the DPPH scavenging activitiy; 27.95%, 28.68%, and 12.61% of the Fe2+ chelating ability; and the absorbance value of the reducing power (OD700nm) were 0.289, 0.385, and 0.557. In anti-inflammatory assays, the cell viability significantly improved to 60.32-70.29% and 64.10-64.84%, respectively after treated in co-treatment and post-induced with 2 g/mL FOS3k and 2 ng/mL FX on lipopolysaccharides (LPS) induced RAW264.7 cell, while compared to that of the control group (50.14%) (p < 0.05). Compared to control group (24.05 mM), the NO release content significantly decreased 34.73% and 16.17% after treated in co-treatment with 2 g/mL FOS3k and 4 ng/mL FX on lipopolysaccharides (LPS) induced RAW264.7 cell (p < 0.05). FOS3k have anti-inflammatory activity on lipopolysaccharides (LPS) induced RAW264.7 cell when treated in co-treatment with 0.02 g/mL concentration, it can significantly decrease the secretion of TNF-38.63 ng/mL, IL-6 (160.29 ng/mL), IL-10 (8.76 ng/mL) while compared to that of the control group (56.52, 211.02, 25.31 ng/mL) (p < 0.05). Compared to the control group (56.52, 211.02, 25.31 ng/mL), it has anti-inflammatory activity on LPS induced RAW264.7 cell while treated in co-treatment with 4 ng/mL FX, which significantly decrease the secretion of TNF-37.05 ng/mL, IL-6 (109.92 ng/mL), IL-10 (16.85 ng/mL) (p < 0.05). Above all, LJ-FOS3k and LJ-FX obtained from this experiment perform antioxidant and anti-inflammatory activity.
| Identifer | oai:union.ndltd.org:TW/107NTOU5253050 |
| Date | January 2019 |
| Creators | Huang, Si-Ying, 黃思穎 |
| Contributors | Pan, Chorng-Liang, 潘崇良 |
| Source Sets | National Digital Library of Theses and Dissertations in Taiwan |
| Language | zh-TW |
| Detected Language | English |
| Type | 學位論文 ; thesis |
| Format | 109 |
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