碩士 / 國立臺灣海洋大學 / 食品科學系 / 97 / This study attempted to know bioactivities of oligosaccharides from Spirulina. Spirulina polysaccharides (spirulan) were degraded by commercial enzymes, Pseudomonas vesicularis MA103-, and Aeromonas salmonicida MAEF108- crude agarases. The changes of reducing sugar content of spirulan hydrolyzed for 24 hr by ��-glucuronidase and industrial cellullase (2.09 mg/mL and 1.99 mg/mL) were higher than alginate lysase, xylanase, cellullase, and ��-glucuronidase. Experimental data showed that spirulan were hydrolyzed by 5 unit/dL MAEF108-crude agarases for 6 hr then hydrolyzed by 5 unit/dL MA103-crude agarases for 18 hr at optimum condition of individual enzyme contented the highest reducing sugar content. Sulfated group content of spirulan hydrolysate was higher than spirulan without hydrolyzed (83.75 �慊/mg), and sulfated group content of spirulan hydrolyzed by MA103-crude agarase was the highest in each group (791.11 �慊/mg). MA103- and MAEF108-crude spirulan inducing enzyme (MA103-SE and MAEF108-SIE) were induced by 0.15% or 0.30% spirulan at 20, 26, and 30oC. The optimum inducing condition of MA103-SIE was induced by 0.15% spirulan at 26oC for 72 hr, and the enzyme activity was 0.31 U (1 �慆ol rhamnose equivalent/mL/min). When induced by 0.15% spirulan at 26oC for 48 hr, the enzyme activity of MAEF108-SIE was 0.32 U. That was the optimum inducing condition. In evaluating scavenging effect on DPPH free radicals, > 5 kDa spirulan oligosaccharides-hydrolysates fraction was had the best (59.14%) in each group. < 3 kDa spirulan oligosaccharides-hydrolysates had the best chelating effects on ferrous ions (86.4%). Anticoagulant activity was analyzed by activated partial thromboplastin time (APTT) and prothrombin time (PT) of human plasma. APTT of 3-5 kDa fraction (200 �慊/mL) was longer than 3-5 kDa fraction (100 �慊/mL), but there was no significantly different between control group. It was similar to < 3 kDa fraction. All of the spirulan oligosaccharides-hydrolysates were no extension for PT, but their optical density were lower than control group. Cell study showed that 3-5 kDa and < 3 kDa spirulan oligosaccharides-hydrolysates had significantly increased cell viability of mouse embryonic fibroblast NIH 3T3 cell for 48 and 72 hr culturing in the concentration of 3.9 to 2000 �慊/mL. 3-5 kDa spirulan oligosaccharides-hydrolysates showed proliferation effect on human skin fibroblast CCD-966SK. The concentrations were 62.5 to 2000 �慊/mL had increased cell viability more than 115%.s Cell viability of < 3 kDa (2000 �慊/mL) was 127.9%. The result showed they enabled to proliferate NIH 3T3 and CCD-966SK cell. For melanoma cell (A375) growth inhibition experiments, it was found that 3-5 kDa spirulan oligosaccharides-hydrolysates inhibited A375 cell proliferation by 80.8% at 24hr, when the concentration was 250 �慊/mL. Cell viability of 125 �慊/mL 3-5 kDa spirulan oligosaccharides-hydrolysates was 33.2%. Inhibition of < 3 kDa spirulan oligosaccharides-hydrolysates was lower than 3-5 kDa. There was no significant tyrosinase inhibition of 3-5 kDa and < 3 kDa spirulan oligosaccharides-hydrolysates. Spirulan oligosaccharides-hydrolysates showed antioxidatnt activity, A375 cell inhibition effect, and CCD-966SK cell proliferation effect, thus potential to develop novel health food or skin care products.
| Identifer | oai:union.ndltd.org:TW/097NTOU5253070 |
| Date | January 2009 |
| Creators | Tsen-Yin Lin, 林岑穎 |
| Contributors | Chorng-Liang Pan, 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 | 102 |
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