碩士 / 國立臺灣海洋大學 / 食品科學系 / 106 / This study is aim to analyze characteristic and bioactivities of ulvan oligosaccharides produced by enzymatic hydrolysis and investigate conditions of lactic acid fermentation of Ulva residue. Firstly, ulvan were gained from Ulva lactuca powder that treated by hot-water (110oC/75 min), then collected by ultrafiltration system (> 30 kDa), and precipitated in 95% ethanol. The yield of ulvan is 15.89% (15.89 g of ulvan obtained from 100 g of Ulva lactuca powder), and the total sugar, reducing sugar, uronic acid, sulfate, protein and total phenols of ulvan are 48.59%, 3.05%, 30.22%, 21.29%, 0.47% and 0.11%, respectively. HPLC chromatogram is shown molecular weights (MW) of ulvan are mainly 769 kDa polysaccharides which relative ratio of fractions are 53.2%. FT-IR spectra of ulvan is presented O-H, C=O, C-O and S=O stretching. Moreover, using ulvan to induce marine bacteria Pseudomonas vesicularis MA103 and Aeromonas salmonicida MAEF108 could produce ulvan-degrading enzymes. Activities of ulvan lyase, amylase, cellulase, and xylanase were 1.93 U/mL, 3.78 U/mL, 2.72 U/mL, and 1.62 U/mL respectively which produced by MA103 in 3 days, and were 0.22 U/mL, 0.17 U/mL, 0.19 U/mL, and 3.23 U/mL respectively which produced by MAEF108 in 2 days. Further, ulvan were hydrolyzed by crude enzymes solution of MA103 and MAEF108, and differentiated by ultrafiltration system (< 3 kDa) to acquire ulvan oligosaccharides (UOS3K). The yield of UOS3K is 4.69% (4.69 g of ulvan gained from 100 g of Ulva lactuca powder), and the total sugar, reducing sugar, uronic acid, sulfate and total phenols of ulvan oligosaccharides are 7.42%, 7.17%, 2.54%, 13.19% and 0.73%, respectively. HPLC chromatogram is shown MW of UOS3K are mainly 800 Da oligosaccharides which relative ratio of fractions are 82.0%. FT-IR spectra of UOS3K is presented O-H, C=O, C-O and S=O stretching. Secondly, the bioactivity of ulvan or UOS3K were evaluated by angiotensin I converting enzyme (ACE) inhibitory, antioxidant activities and anticoagulant activities in vitro. In ACE inhibitory assay (ACEI), inhibition of ulvan (50 mg/mL) and UOS3K (12.5 mg/mL) were 18.41% and 81.86%. IC50 value of UOS3K on ACEI was 0.451 mg/mL (Peptide concentration). In antioxidant activity assays, the DPPH scavenging activity of ulvan (10 mg/mL) and UOS3K (10 mg/mL) were 52.46% and 27.27 %; the Fe2+ chelating ability of ulvan (20 mg/mL) and UOS3K (10 mg/mL) were 4.62% and 97.98%; both of ulvan and UOS3K were not shown the reducing power. In anticoagulant activity assays, wich are activated partial thromboplastin time (APTT), prothrombin time (PT) and thrombin time (TT) using rabbit plasma, UOS3K did not prolong the aggregation time compared to blank group. However, UOS3K could decrease the aggregation level to 94% and 78% in APTT and PT test. Thirdly, utilized Ulva residue which removed the ulvan from Ulva lactuca to induce MA103 and MAEF108 producing crude enzymes was investigated. Activities of ulvan lyase, amylase, and xylanase were 1.97 U/mL, 3.91 U/mL, and 2.08 U/mL respectively which produced by MA103 in 3 days, and activities of enzymes produced by MAEF108 were no significant. Additionally, hydrolyzing conditions of the Ulva residue were evaluated. Finally, Ulva residue polysaccharide hydrolysate was added 0.5% (w/v) yeast extract as nitrogen source, calcium carbonate as neutralizing agent, and then fermented by 1% (v/v) of Lactobacillus plantarum BCRC12327 at 30oC for 36 hr, the lactic acid concentration was 35.97 g/L and lactic acid yield was 15.65% (15.65 g of lactic acid gained from 100 g of Ulva residue).
| Identifer | oai:union.ndltd.org:TW/106NTOU5253060 |
| Date | January 2018 |
| Creators | Hung, Yueh-Hao, 洪悅豪 |
| 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 | 111 |
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