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  • 學位論文

西洋參渣生物轉化物之非消化醣類性質與結構特徵

Properties and structure characterization of indigestible carbohydrates from the bioconversion product of American ginseng residues

指導教授 : 呂廷璋
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摘要


已知西洋參與靈芝多醣具許多免疫調節之活性,工業生產之西洋飲品可以是生產靈芝與西洋參渣多醣之原料。本研究使用西洋參渣靈芝菌絲發酵轉化物 (轉化物) 為原料,研究此轉化物之多醣性質與活性,並比較西洋參多醣活性區分之多醣性質。西洋參非消化性水溶性多醣區分具有豐富之Arabinogalactan II (AGII) 與少量Arabinogalactan I (AGI)、Rhamnogalcturonan I (RGI) 與RhamnogalcturonanII (RGII)之結構特徵,推測以西洋參之AG II貢獻刺激RAW 264.7釋放NO或TNF-α之效果。轉化物以熱水萃取、酒精沉降與酵素水解後,藉由酒精沉降分成非消化水溶性多醣層 (IDPS) 與非消化短鏈醣層 (SCC)。IDPS經電荷層析可分成5個區分 (IDPS F1、F2、F3、F4與F5)。IDPS F1有豐富之 (1,3;1,6)-β-D-glucan 與少量之Xyloglucan與AG II;IDPS F2有 (1,3;1,6)-β-D-glucan、RG I、AG I、AGII;IDPS F3有β–glucan、RG I、AG I、AG II、Arabinan、RG I與Homogalacturonan (HG);IDPS F4與F5以RG I、AG I、AG II、Arabinan、RG I與HG為主要之醣結構。SCC經電荷劃分可分成3個區分 (SCC F1、SCC F2與SCC F3) 與其中SCC F1經分子篩層析可分成5個中性醣區分 (SCC F1A、F1B、F1C、F1D、F1E)。SCC F1A有Galactomannan、Xyloglucan、AG I、AG II與Arabinan;SCC F1B有(1,6)-glucan、(1,4)-Mannan;SCC F1C、F1D、F1E分別為麥芽三糖、麥芽二糖與葡萄糖為主要組成;SCC F2有RG I、AGI、Arabinan與β–glucan;SCC F3有RG I、Arabinan、AG II、Arabinogalactan protein。評估各區分刺激RAW 264.7釋放NO與TNF-α活性。IDPS F2、F3、F4、F5具刺激RAW 264.7釋放TNF-α活性,但不具調節NO釋放效果,其中IDPS F2以 (1,3;1,6)-β-D-glucan 為主要活性貢獻之來源。SCC F1、F2、F3皆具有調節RAW 264.7釋放NO之活性,但僅SCC F1、F2具刺激RAW 264.7釋放TNF-α之活性。測試SCCF1中之活性成分,發現以SCCF1A為主要活性區分之貢獻。西洋參渣經靈芝轉化可釋出果膠多醣與半纖維素,並產生免疫調節活性之β-glucan。

並列摘要


The immune modulatory activity of polysaccharides from America ginseng and Ganoderma lucidum has been well understood. The industrial by-product of America ginseng drinks can be used as the raw material to produce the Ganoderma lucidum bioconversion product of American ginseng residues. In the research, the bioconversion product and the America ginseng bioactive polysaccharide were investigated. The America ginseng bioactive polysaccharide is rich in arabinogalactan II ( AG II) and a little arabinogalactan I (AGI), rhamnogalacturonan I (RGI) and rhamnogalcturonanII (RGII). It is presumed that the stimulation effect on RAW264 of the American ginseng polysaccharides was contributed by AG II. After the bioconversion product was treated by hot water extraction, ethanol precipitation and enzyme digestion, it could be divided into indigestible polysaccharides (IDPS) and indigestible short chain carbohydrate (SCC). IDPS could be separated by DEAE chromatography into 5 fractions, IDPS F1, F2, F3, F4 and F5. IDPS F1 has the characterization of (1,3;1,6)–β–glucan, AGII, and xyloglucan. IDPS F2 has the characterization of (1,3;1,6)–β–glucan, RG I, AG I, and AGII. IDPS F3 has the characterization of β–glucan, AGI, AGII, arabinan, RGI, RGII, and homogalacturonan (HG). IDPS F4 and F5 has the characterization of AGI, AGII, arabinan, RGI, RGII, and HG. SCC can be separated by DEAE chromatography into 3 fractions, SCC F1, F2, and F3. Moreover, SCC F1 could be divided by size exclusion chromatography into SCC F1A, F1B, F1C, F1D, and F1E. SCC1A has the characterization of galactomannan, AGI, AGII, arabinan, and xyloglucan. SCC F1B has the characterization of (1,6)-glucan and (1,4)-mannan. SCC F1C, F1D, and F1E are the maltotriose and maltobiose and glucose individually. SCC F2 contains the structure of RG I, AG I, arabinan, and β–glucan. SCC F3 has the characterization of RGI, arabinan, AG II, arabinogalactan protein, and RGII. According to RAW 264.7 cell model, IDPS F2, F3, F4, F5 could stimulate RAW 264.7 TNF-α production but did not regulate the release of NO. Furthermore, (1,3;1,6)-β-glucan in IDPS F2 is the main activity structure. SCC F1, F2, and F3 could stimulate the release of NO from RAW 264.7 but only SCC F1 and F2 stimulated the release of TNF-α. SCCF1A is the main contribution of the immunomodulatory activity to SCC F1. After Ganoderma lucidum fermented the America ginseng residue, it can not only release pectin polysaccharides and hemicellulose but also produce β-glucan which have the immunomodulatory activity.

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