:::

| | | | | 第 | | 筆

詳目顯示區塊

第 1 筆
| (以作者名查詢海洋大學圖書館館藏系統)
| (以作者名查詢全國圖書書目資訊網)
| (以作者名查詢全國博碩士系統)
論文名稱: | Pseudomonas vesicularis MA103 Fucosidase 之 fucI基因轉殖至 Lactococcus lactis NZ3900 表現及Fucosidase rfucI-H水解岩藻聚醣所得寡醣於 Caco-2 細胞吸收效果之探討
| Studies on Cloning and Expression of Pseudomonas vesicularis MA103 Fucosidase fucI Gene on Lactococcus lactis NZ3900 and Evaluation the Adsorption Effect of Oligosaccharides from Hydrolyzed Fucoidan by Fucosidase rfucI-H from on Caco-2 Cells
研究生: | 朱美嘉 Mei-Chia Chu
指導教授: | 潘崇良 Chorng-LiangPan
| 張君如 Chun-Ju Chang
學位類別: | 碩士
學校名稱: | 國立臺灣海洋大學
系所名稱: | 食品科學系
學號: | 19932063
畢業學年度: | 100
語文別: | 中文
關鍵字: | 岩藻聚醣 Fucoidan
| 乳酸鏈球菌 NZ3900 Lactococcus lactis NZ3900
| Caxo-2細胞 Caco-2 cell
全文影像連結: | | (本論文20150730對校內公開)
| 論文全文(pdf)

論文頁數: | 132
摘要: | 本研究目的在探討Pseudomonas vesicularis MA103 fucI基因構築至乳酸菌 (Nisin controlled expression system, NICE) 表現系統之 pNZ8149載體上，再以電穿孔法轉殖至 Lactococcus lactis NZ3900 中，以 Nisin 誘導該轉殖株產 Fucosidase 並降解莢托馬尾藻 (Sargassum siliquosum) 之岩藻聚醣 (Fucoidan) ，探討並比較莢托馬尾藻 Fucoidan (SSF) 及其衍生寡醣之抗凝血、抗氧化之生理活性，並評估岩藻寡醣於 Caco-2 細胞吸收之效果。依據菌株 MA103 中 Fucosidase 之 fucI 基因與乳酸菌 pNZ8149限制酶切位，設計引子對分別於 5’ 和 3’ 端加入 SpeI 和 SacI 限制酶切位，另設計一組接上His-tag之引子對。電穿孔轉形至 E. coli DH5α，抽取含 fucI基因之質體與乳酸菌表現載體 pNZ8149 接合，電穿孔轉形至 Lc. lactis NZ3900 之勝任細胞中表現。乳酸菌轉殖株 Lc. lactis NZ3900 rfucI 及 Lc. lactis NZ3900 rfucI-H 以 5 ng/mL Nisin 誘導 3 hr 之胞內酵素活性最佳，分別可得0.056 U/mg protein 及 0.057 U/mg Protein比活性，經 SDS-PAGE 可觀察推估分子量約為 51 kDa之條帶。另外，將 SSF進行高效能液相層析 (High performance liquid chromatograph, HPLC)，與標準品比對，推估其平均分子量約為 25,000 Da，以Lc. lactis NZ3900 rfucI-H胞內粗酵素液 (Fucosidase rfucI-H) 水解後利用超過濾系統區分出小於 3 kDa分子量之寡糖，經過 Sephadex G-10膠體過濾層析 (Gel permeation chromartography, GPC) 後，以HPLC 分析並比對標準品得知，可能是聚合度為 DP3 之寡醣 (Fuc-C3)，硫酸基含量為 0.28 ± 0.14%。在兔血漿在抗凝血實驗中，於活化部分凝血活酶時間 (Activated partial thromboplastin time, APTT) 實驗中，SSF濃度為 5 mg/mL 時，其 APTT 延遲效果等同於 10 g/mL 肝素與空白組相比可延遲 88 sec，而凝集程度則較 10 g/mL 肝素低， Fuc-C3 濃度則至 20 mg/mL 時才具有延遲 APTT 之效果，與空白組相比可延遲 66 sec，無降低凝集程度之能力。而在凝血酶原時間 (Prothrombin time, PT) 實驗中，結果顯示 SSF 及 Fuc-C3 皆無法延遲凝血時間，但濃度為 5 mg/mL 時具有降低凝血程度之效果。測試 SSF及 Fuc-C3 之抗氧化活性，SSF 有較佳的清除 DPPH自由基能力 (27.12 ± 0.01%) 及還原力 (OD700nm = 0.21 ± 0.01)，Fuc-C3 則有較佳的螯合亞鐵能力 (24.29 ± 0.02%)。在人類腸道上皮細胞 Caco-2 吸收實驗上，以 HPLC 分析，可發現隨著吸收時間的增加，上層液 Fuc-C3 之波峰有下降之趨勢，但在下層液部分並未出現其波峰，進一步使用 TLC 確認，當吸收時間達 90 min時，可在下層液中觀察到 Fuc-C3 色斑之產生，測定其全糖量定量，初步推估其吸收率為 0.1%，而其吸收機制與反應條件仍需進一步探討。
| The aim of this study is using fucI gene from Pseudomonas vesicularis MA103 construct on LAB nisin controlled expression (NICE) system vector, pNZ8149, then cloned into Lactococcus lactis NZ3900 by electroporating and using nisin induced recombinant crude enzyme to hydrolyzed fucoidan from Sargassum siliquosum. Contrast the effects of Sargassum siliquosum fucoidan (SSF) and fucoidan oligosaccharides on their anticoagulant, antioxidant activities, and its adsorption performance with intestinal cell line Caco-2 are examined. The fucI forward primer encode SpeI restriction site and the reverse primer encode SacI restriction site, and one has his-tag. Then using the gene of Pseudomonas vesicularis MA103 fucosidase fucI gene as a template for PCR reaction, and then transformed into the competent cells of E. coli DH5α. Then pGEM-T vector, which carrying the cloned fucI gene, was digested with SpeI and SacI, and ligated into the LAB vector pNZ8149 and electroporated into the Lc. lactis NZ3900. The clones Lc. lactis NZ3900 rfucI and Lc. lactis NZ3900 rfucI-H induced by 5 ng/mL nisin at 30oC for 3 hr could obtain higher fucosidase activity (0.056 U/mg protein and 0.057 U/mg), which revealed a 51 kDa protein band by SDS-PAGE. Using high performance liquid chromatograph (HPLC) to analysis SSF compare to standard, the average molecular weight was 25,000 Da. The SSF hydrolyzed by Lc. lactis NZ3900 rfucI-H intracellular crude enzyme (Fucosidase rfucI-H) and molecular mass < 3 kDa were collected by ultra filtration (UF) system, then though gel permeation chromartography (GPC) and analysis by HPLC. Compare to standard, the DP of fucoidan oligosaccharides may DP 3 (Fuc-C3). In the experiment of anticoagulant activities of rabbit plasma, activated partial thromboplastin time (APTT) analysis, 5 mg/mL of SSF can delayed 88 sec like 10 g/mL heparin, and also reduction aggregation level better than 10 g/mL heparin. The 20 mg/mL of Fuc-C3 can delayed 66 sec. The anticoagulant activities of rabbit plasma in prothrombin time (PT) analysis were examined, SSF and Fuc-C3 were no obvious delayed in prothrombin time, but there were a tendency for reduction on absorbance while concentration were 5 mg/mL. The experiment on antioxidant activity of SSF and Fuc-C3, SSF has better DPPH scavenging effect (27.12 ± 0.01%) and reducing power (OD700nm = 0.21 ± 0.01), and Fuc-C3 has better Fe2+ chelating effect (24.29 ± 0.02%). In the adsorption performance with Caco-2 cells on transwell device, the HPLC analysis showed that Fuc-C3 did not penetrate to the basolateral liquid but the Fuc-C3 concentration were decreased in apical fluid in a time-dependent fashion. Using thin layer chromatography (TLC) to confirm intestinal absorption, Fuc-C3 was observed in basolateral fluid after 90 min. Further measurement on total suger content and the absorption rate of Fuc-C3 was estimated as 0.1%. The possible absorption mechanism and reaction conditions of Fuc-C3 by intestinal cell need further investigation.
|
|
論文目次: | 目錄    i
| 表目錄    vi
| 圖目錄    vii
| 附錄目錄    x
| 中文摘要    xi
| 英文摘要 (Abstract)    xiii
| 壹、前言        1
| 貳、文獻整理        3
| 一、海藻        3
| 二、海藻多醣        4
| 2-1. 海藻多醣定義        3
| 2-2. 海藻多醣之萃取方式        4
| 2-3. 海藻多醣之生理活性        5
| 三、海藻寡醣        6
| 3-1. 海藻寡醣之萃取方法        6
| 3-2. 海藻寡醣之生理活性        7
| 四、岩藻聚醣 (Fucoidan)        9
| 4-1. 岩藻聚醣的組成與結構        9
| 4-2. 岩藻聚醣之生理活性        10
| 五、岩藻醣苷酶 (Fucosidase)        12
| 六、乳酸菌        12
| 6-1. 定義        12
| 6-2. 基因轉殖之應用        13
| 七、Caco-2 細胞        13
| 7-1. 腸道吸收模式    14
| 7-2. 寡醣之吸收    15
| 参、實驗設計        16
| 肆、材料方法        18
| 一、實驗材料        18
| 1-1. 原料        18
| 1-2. 實驗菌株        18
| 1-3. 實驗細胞株        18
| 1-4. 試驗藥品        18
| 1-4-1.  藥品        18
| 1-4-2.  細胞培養液        20
| 1-4-3.  載體        21
| 1-4-4.  引子對 (Primer pair)        21
| 1-4-5.  聚合酶連鎖反應試劑        21
| 1-4-6.  限制酶        21
| 1-4-7.  蛋白質及DNA萃取純化套裝試劑        21
| 1-4-8.  電泳標準品        21
| 1-4-9.  培養基組成        22
| 1-4-10. 細胞培養液組成        23
| 1-4-11. Fucosidase 反應基質        24
| 1-4-12. 電泳溶液        24
| 1-4-13. 電泳膠片配製        24
| 1-4-14. 膠過濾緩衝液        24
| 1-4-15. 無菌過濾        25
| 1-4-16. 薄層層析矽膠片        25
| 1-5. 儀器設備        25
| 二、實驗方法        27
| 2-1. 菌株保存與活化        27
| 2-1-1.  菌株之保存        27
| 2-1-2.  菌株之活化        27
| 2-2.  P. vesicularis MA103 之fucI 基因選殖至乳酸菌        28
| 2-2-1.  引子設計        28
| 2-2-2.  PCR擴增欲用於TA-cloning 之基因        28
| 2-2-3.  大腸桿菌之 TA-cloning 轉形作用        29
| 2-2-3-1. 連接反應        29
| 2-2-3-2. 大腸桿菌 (E. coli DH5) 勝任細胞製作        30
| 2-2-3-3. 大腸桿菌 (E. coli DH5) 電穿孔轉形        30
| 2-2-3-4 利用膠電泳確認插入之基因片段大小        30
| 2-2-3-5. 利用限制酶剪切純化目標基因        31
| 2-2-4. 乳酸菌之電穿孔轉形作用        31
| 2-2-4-1. 連接反應        31
| 2-2-4-2. 乳酸菌 (Lc. lactis NZ3900) 勝任細胞之製備        32
| 2-2-4-3. 乳酸菌 (Lc. lactis NZ3900) 電穿孔轉形        33
| 2-2-4-4 利用膠電泳確認插入之基因片段大小        33
| 2-2-4-5. 利用限制酶剪切純化目標基因        34
| 2-2-5. DNA 序列比對        34
| 2-2-6. Nisin對重組乳酸菌生長之影響        34
| 2-2-7. 乳酸菌重組 Fucosidase之酵素活性分析        34
| 2-2-7-1. Fucosidase 活性測定        34
| 2-2-7-2. Lc. lactis NZ3900 rpfuI 之Nisin 誘導濃度對Fucosidase 活性碳討        35
| 2-2-7-3. 蛋白質的定量        35
| 2-2-7-4. 分子量的鑑定        36
| 2-3. 莢托馬尾藻一般成份分析        36
| 2-3-1.  水分含量        36
| 2-3-2.  灰分含量        36
| 2-3-3.  粗蛋白含量        36
| 2-3-4.  粗脂肪含量        37
| 2-3-5.  粗纖維含量        37
| 2-4.岩藻聚糖之萃取及其經 Fucosidase rfucI-H 水解後寡醣產物之分析        38
| 2-4-1.  岩藻聚醣之萃取        38
| 2-4-2.  總醣量測定        38
| 2-4-3.  還原醣量測定        38
| 2-4-4.  硫酸酯含量測定        39
| 2-4-5.  總酚含量測定        39
| 2-4-6.      Fucosidase rfucI-H 之製備        39
| 2-4-7.  岩藻寡醣之製備        40
| 2-4-8.  膠體過濾層析 (GPC)        40
| 2-4-9.  超過濾 (Molecular weight cut off, MWCO)        40
| 2-4-10. 薄層層析 (TLC)        41
| 2-4-11. 高效能液相層析 (HPLC)        41
| 2-4-12. 傅立葉紅外線光譜分析 (FT-IR)        41
| 2-5. 抗氧化活性測定        41
| 2-5-1.  清除 DPPH 自由基能力測定        41
| 2-5-2.  還原能力之測定        42
| 2-5-3.  亞鐵離子螯合能力之測定        42
| 2-6. 抗凝血生理活性        42
| 2-6-1.  凝血酶原時間 (Prothrombin time, PT)        42
| 2-6-2.  活化部份凝血活酶時間 (Activited
| partial thromboplastin time, APTT)        43
| 2-7. 岩藻寡醣於腸道細胞吸收試驗        43
| 2-7-1. Caco-2 細胞株之活化        43
| 2-7-2. Caco-2 細胞株培養方法及繼代方法        44
| 2-7-3. 細胞數計數        44
| 2-7-4. 細胞之冷凍保存        44
| 2-7-5. Caco-2 細胞存活率分析-MTT assay        45
| 2-7-5-1. 以MTT assay測定不同細胞數對 Caco-2
| 細胞存活率之影響    45
| 2-7-5-2. 岩藻寡醣對 Caco-2細胞之毒性試驗    46
| 2-7-6. Transwell insert 培養法        47
| 2-7-6-1. Caco-2 細胞於吸收裝置其單層膜完整性測試    47
| 2-7-7. 岩藻寡醣於腸道細胞吸收率測定        47
| 三、統計分析        47
| 伍、結果與討論        48
| 陸、結論        61
| 柒、參考文獻        62
|
參考文獻: | 卡伯特。2010。低聚合度洋菜寡糖的鑑定和應用以及洋菜多糖降解機制的探討。國立臺灣海洋大學食品科學系博士學位論文。基隆。臺灣。
| 江仁淳。2001。臺灣重緣葉馬尾藻海藻多醣體之免疫調節功能評估。國立臺灣海洋大學食品科學系碩士學位論文。基隆。臺灣。
| 呂廷璋。1986。礁膜 (Monostroma nitidum) 水溶性多醣的理化性質。國立臺灣海洋學院水產食品科學研究所碩士學位論文。基隆，
| 臺灣。
| 吳紹祺。1999。海洋菌所產 agarase 生產條件與海洋多醣經 agarase 分解所得寡醣類組成之探討。國立臺灣海洋大學食品科學系碩士學位論文。基隆。臺灣。
| 吳紹祺。2005。海洋多醣洋菜酶水解物及其發酵產物之生理活性研究。國立臺灣海洋大學食品科學系博士學位論文。基隆。臺灣。
| 李冠青。2008。食品多醣、甲基化衍生物及降解物之抗氧化活性及
| 對腸腺癌細胞之抑制性。靜宜大學食品營養學系碩士學位論文，臺中，臺灣。
| 林季磊。2011。海藻寡醣經分離與提高硫酸基含量後之生理活性探討。國立臺灣海洋大學食品科學系碩士學位論文。基隆。臺灣。
| 胡幼圃。2000。美國藥學科學家會議報告，科學發展月刊。29: 283-285.
| 洪啟芳。2003。硫化程度對一些多醣及寡醣之抗凝血性質之探討。
| 大同大學生物工程研究所碩士學位論文。臺北。臺灣。
| 高慶煌。2002。以熱分析法測定基丁質類物質之乙醯化程度。國立
| 臺灣海洋大學食品科學系碩士學位論文，基隆。臺灣。
| 郭馨鎂。2006。海藻寡糖對雙叉桿菌之助生性及在發酵乳應用之研究。國立臺灣大學食品科技研究所碩士學位論文。臺北，臺灣。
| 陳怡均。2001。Galectin-1 與Thioredoxin peroxidase II 基因之選殖及表現。國立中央大學生命科學系碩士學位論文。桃園。臺灣。
| 陳柏璇。2010。Aeromonas salmonicida MAEF108 之 Agarase AS-IIIb 基因轉殖至乳酸菌及乳酸菌轉殖株發酵山藥乳製品之探討。國立臺灣海洋大學食品科學系碩士學位論文。基隆。臺灣。
| 陳錫秋、高泉創。1983。青海菜及carrageenan對降低膽固醇之效果。中國水產。371: 5-13。
| 陳慶源、林富美。2004。益生菌之保健功能。食品工業。36: 1-3。
| 馮家輝、鍾佳華、李欣翰、卡伯特 (B. Kazlowski)、潘崇良。2007。酸和agarases組合降解龍鬚菜多醣液所得寡醣之組成分析，臺灣食品科學技術學會年會，2007年12 月07 日，彰化，B-57。
| 黃淑芳。1989。認識藻類。臺灣省立博物館出版。台北市。pp. 1-54。
| 黃霨禎。2005。龍鬚菜寡糖水解物之製備及其抗氧化與抗至突變性之探討與對雙叉桿菌生長影響。國立臺灣海洋大學食品科學系碩士學位論文。基隆。臺灣。
| 葉美娟、許家榮。2005。以葡萄醣醛酸酶基因gusA 為報告基因探討 nisin 調控表現系統於乳酸菌 Lactobacillus paracasei 之調控表現研究。臺灣農業化學與食品科學。43: 86-96。
| 楊依珊。2011。Pseudomonas vesicularis MA103 所產 Alginate lyase 之基因轉殖至 Lactococcus lactis NZ3900表現及 Alginate lyase 所產寡醣產物分析和用於酵母菌發酵生產乙醇之探討。國立臺灣海洋大學食品科學系碩士學位論文。基隆。臺灣。
| 廖啟成。1998。乳酸菌之分類應用。食品工業。30: 1-10。
| 潘崇良、陳衍昌、 蔡震壽、方翠筠、黃意真、鄒文雄、林富邦、劉秀美、唐世杰。2009。海洋功能性醣體研究: 藻類多醣生質乙醇與丁醇及碳交易附加收入。海洋功能性醣體研究: 藻類多醣生質乙醇與丁醇及碳交易附加收入。第三屆海峽兩岸「魚類生理與養殖」研討會，國立臺灣海洋大學水產生物科技頂尖研究中心主辦，行政院國家科學委員會、中央研究院細胞與個體生物學研究所協辦，2009年10月19-21日。基隆。臺灣。
| 劉綉婷。2008。Pseudomonas vescicularis MA103 菌株所產 Agarase PV-1b 純化與基因選殖至大腸桿菌表現之探討。國立臺灣海洋大學食品科學系碩士學位論文。基隆。臺灣。
| 劉錦芬。2001。臺灣養殖龍鬚菜萃取物之免疫生理活性探討。國立臺灣海洋大學食品科學系碩士學位論文。基隆，臺灣。
| AOAC. 1997. Offical Methods of Analysis.  16th edition.  Association of Official Analytical Chemists.  Washington, DC, U.S.A.
| Adhikaria, U., Mateub, C. G., Chattopadhyaya, K. C., Pujolb, A., Damonteb, E. B., and Ray, B. 2006. Structure and antiviral activity of sulfated fucans from Stoechospermum marginatum. Phytochemistry 67: 2474-2482.
| Ale, M. T., Maruyama, H., Tamauchi, H., Mikkelsen, J. D., and Meyer, A. S. 2011. Fucoidan from Sargassum sp. and Fucus vesiculosus reduces cell viability of lung carcinoma and melanoma cells in vitro and activates natural killer cells in mice in vivo. International Journal of Biological Macromolecules 49: 331- 336.
| Anggadiredja, J., Andyani, R., and Muawanah, H.  1997. Antioxidant activity of Sargassum polycystum (Phaeophyta) and Laurencia obtuse (Rhodophyta) from Seribu Islands. Journal of Applied Phycology  9: 477-479.
| Aspinall, G. O. 1982.  Isolation and fractionation of polysaccharides. In: The Polysaccharides. Aspinall, G. O. Academic Press Inc., New York , U.S.A.. Ch 1, pp. 19-34.
| Athukorala, Y., Lee, K. W., Kim, S. K., and Jeon, Y. J.  2007.  Anticoagulant activity of marine green and brown algae collected from Jeju Island in Korea. Biotechnology  98: 1711-1716.
| Athukorala, Y., Jung, W. K., Vasanthan, T., and Jeon, Y. J. 2006.  An anticoagulative polysaccharide from an enzymatic hydrolysate of Ecklonia cava.  Carbohydrate Polymers  66: 184-191.
| Balimane, P. V., and Chong, S. 2005. Cell culture-based models for intestinal permeability : A critique. Drug Discovery Today 10: 335-343.
| Bertani, G. 1951. Studies on lysogenesis. I. The mode of phage liberation by lysogenic Escherichia coli. The Journal of Bacteriology  62: 293-300.
| Berteau, O., and Mulloy, B. 2003. Sulfated fucans, fresh perspectives: Structures, functions, and biological properties of sulfated fucans and an overview of enzymes active toward this class of polysaccharide. Glycobiology 13: 29-40.
| Brasch, D. J., Chang, H. M., Chuah, C. T., and Melton, L. D. 1981.  The galactan sulfate from the edible, red alga Porphyra columbina.  Carbohydrate Research  97: 113-125.
| Braun, A.,  Hammerle, S.,  Suda, K., Rothen-Rutishauser, B., Gunthert, M., Kramer, S. D., Wunderli-Allenspach, H. 2000. Cell cultures as tools in biopharmacy. European Journal of Pharmaceutical Sciences 11: 51-60.
| Bilan, M. I., Grachev, A. A., and Ustuzhanina, N. E. 2002. Structure of a fucoidan from the brown seaweed Fucus evanescens C.Ag. Carbohydrate Research 337: 719-730.
| Black, W. A. P., Dewar, E. T., and Woodward, F. N. 1952. Manufacture of algal chemicals. IV-laboratory-scale isolation of fucoidin from brownmarine algae. Journal of the Science of Food and Agriculture 3: 122-129.
| Carlucci, M. J., Pujol, C. A., Ciancia, M., Noseda, M. D., Matulewicz, M. C., Damonte, E. B., and Cerezo, A. S. 1997. Antiherpetic and anticoagulant properties of carrageenans from the red seaweed Gigartina skottsbergii and their cyclized derivatives: Correlation between structure and biological activity. International Journal of Biological Macromolecules  20: 97-105.
| Chen, H., Yan, X., Zhu, P., and Lin, J. 2006. Antioxidant activity and hepatoprotective potential of agaro-oligosaccharides in vitro and in vivo.  Nutrition Journal  5: 31-43.
| Chen, S., Wang, W., Liu, H., Li, C., and Liu, C. 2010. Purificationandlowinghyperlipi-demia activity of fucoidan from Sargassum henslowianum. Food and Fermentation Industries 36 28-31.
| Chizhov, A. O., Dell, A., and Morris, H. R. 1999. A study of fucoidan from the brown seaweed Chorda filum. Carbohydrate Research 320: 108-119.
| Cho, S., Kang, S., Cho, J., Kim, A., Park, S., Hong, Y. 2007. The antioxidant properties of brown seaweed (Sargassum siliquastrum) extracts. Journal of Medicinal Food 10: 479-485.
| Cumashi, A., Ushakova, N. A., Preobrazhenskaya, M. E., D'Incecco, A., Piccoli, A., Totani, L., Tinari, N., Morozevich, G. E., Berman, A. E., Bilan, M. I., Usov, A. I., Ustyuzhanina, N. E., Grachev, A.A., Sanderson, C. J., Kelly, M., Rabinovich, G. A.,  Iacobelli, S., and Nifantive, N. E. 2007. A comparative study of the  anti-inflammatory, nticoagulant, antiangiogenic, and antiadhesive activities of  nine different fucoidans from  brown  seaweeds.  Glycobiology 17: 541-552.
| Cory, A. H., Owen, T. C., Barltrop, J. A., and Cory, J. G. 1991. Use of an aqueous soluble tetrazolium/formazan assay for cell growth assays in culture. Cancer Communication 3: 207-212.
| Daniel, R., Berteau, O., Jozefonvicz, J., and Goasdoue, N. 1999. Degradation of algal (Ascophyllum nodosum) fucoidan by an enzymatic activity contained in digestive glands of the marine mollusk Pecten maximus. Carbohydrate Research 322: 291-297.
| Dawes, C. J. 1998. Marine Botany. John Wiley and Sons, Inc., New York, U.S.A, p. 480.
| de Ruyter, P. G., Kuipers, O. P., Beerthuyzen, M. M., Alen-Boerrigter, I., and de Vos, W. M. 1996. Functional analysis of promoters in the nisin gene cluster of Lactococcus lactis. The Journal of Bacteriology  178: 3434-3439.
| Dodgson, K. S. 1961. Determination of inorganic sulfate in studies on the enzymatic and non-enzymatic hydrolysis of carbohydrate and other sulfate esters. The Journal of Biochemistry  78: 312-319.
| Doh-ura, K., Kuge, T., Uomoto, M., Nishizawa, K., Kawasaki, Y., and Iha, M. 2007. Prophylactic effect of dietary seaweed fucoidan against enteral prion infection. Antimicrobial Agents and Chemotherapy  51(6): 2274-2277.
| Dinis, T. C. P., Madeira, V. M. C., and Almeida, L. M. 1994. Action of phenolic derivatives (acetaminophen, salicylite, and 5-aminosalicylite) as inhibitors of membrane lipid peroxidation and as peroxyl radical scavengers. Archives of Biochemistry and Biophysics  315: 161-169.
| Dubois, M., Gilles, K. A., Hamilton, J. K., Rebers, P. A., and Smith, F., 1956. Colormetric method for determination of sugars and related
| substances. Analytical Chemistry  28: 350-356.
| Duckworth, M., and Yaphe, W. 1970. Thon-layer chromatographic analysis of enzymic hydrolysates of agar. Journal of Chromatography 49: 482-487.
| Federle, M. J., and Bassler, B. L. 2003. Interspecies communication in bacteria.  The Journal of Clinical Investigation  112: 1291-1299.
| Fleurence, J., Coeur, C. L., Mabeau, S., Maurice, M., and Landrein, A. 1995. Comparison of different extractive procedures for proteins from the edible seaweeds Ulva rigida and Ulva rotundata.  The Journal of Applied Phycology 7: 477-582.
| Gan, L. S. L., and Thakker, D. R. 1997. Applications of the Caco-2 Emodel in the design and development of orally active drugs: lucidation of biochemical and physical barriers posed by the intestinal epithelium. Advanced Drug Delivery Reviews 23: 77-98.
| Gnoth, M. J., Rudloff, S., Kunz, C., and Kinne, R. K. H. 2001. Investigations of the in vitro transport of human milk oligosaccharides by a Caco-2 monolayer using a novel high performance liquid chromatography-mass spectrometry technique.  The Journal of Biological Chemistry 276: 34363-34370.
| Gnoth, M. J., Rudloff, S., Kunz, C., and Kinne, R. K. H. 2002. Studies on the intestinal transport of human milk oligosaccharides(HMO) using Caco-2 cells. Food Research International 35: 145-49.
| Hassid, W. Z., 1969. Biosynthesis of oligosaccharides and polysaccharides in plants. Science  165: 137-144.
| Hayashi, T., Hayashi, K., and Kojima, I. 1996. Calcium spirum spirularin, an inhibitor of enveloped virus replication, from a blue-green alga Spirulain platensis. The Journal of Nutrition 59: 83-87.
| Hayashi, K., Nakano, T., Hashimoto, M., Kanekiyo, K., Hayashi, T. 2008. Defensive effects of a fucoidan from brown alga Undaria pinnatifida against herpes simplex virus infection. International Journal of Immunopharmacology  8:109-116.
| Hemmingson, J. A., Falshaw, R., Furneaux, R. H., Thompson, K. 2006. Structure and antiviral activity of the galactofucan sulfates extracted from Undaria pinnatifida (Phaeophyta). Journal of Applied Phycology 18: 185-193.
| Heo, S. J., Jeon, Y. J. Lee, J., Kim, H. T., and Lee, K. W. 2003. Antioxidant effect of enzymatic hydrolyzate from a kelp, Ecklonia cava. Algae 18: 71-81.
| Heo, S. J., Park, E. J., Lee, K. W., and Jeon, Y. J. 2005. Antioxidant activities of enzymatic extracts from brown seaweeds. Bioresource Technology 96: 1613-1623.
| Heyraud, A., Colin-Morel, P., Girond, S., Richard, C., and Kloareg, B. 1996. HPLC analysis of saturated or unsaturated oligoguluronates and oligomannuronates. Application to the determination of the action pattern of Haliotis tuberculata alginate lyase. Carbohydrate Research  29: 115-126.
| Hidalgo, I. J., Raub, T. J, and Borchardt, R. T.  1989. Haracterization of the human colon carcinoma cell line (Caco-2) as a model system for intestinal epithelial permeability. Gastroenterology 96: 736-749.
| Hillgren, K. M., Kato, A., and Borchardt, R.T. 1995. In vitro systems for studying intestinal drug absorption. Medicinal Research Reviews15: 83-109.
| Holtkamp, A. 2009. Isolation, aharacterisation, modification and application of fucoidan from Fucus vesiculosus. Südwestdeutscher Verlag für Hochschulschriften. Stuttgart, Deutschland. pp. 127-128.
| Hu, B., Gong, Q., Wang, Y., Ma, Y.,  Li, J., and Yu, W.  2006.  Prebiotic effects of neoagaro-oligosaccharides prepared by enzymatic hydrolysis of agarose.  Anaerobe  12: 260-266.
| Huang, L., Wen, K., Gao, X., and Liu, Y. 2010. Hypolipidemic effect of fucoidan from Laminaria japonica in hyperlipidemic rats. Pharmaceutical Biology 48: 422-426.
| Hunter, J., Jepson, M. A., Tsuruo, T., Simmons, N. L., and Hirst, B. H.1993. Functional expression of P-glycoprotein in apical membranes of human intestinal Caco-2 cells. The Journal of Bioligical Chemistry 268(20): 14991-14997.
| Hyun. J. H., Kim,S. C., Kang, J. I., Kim,M. K., Boo, H. J., Kwon, J. M., Koh, Y. S., Hyun,J. W., Park, D. B., Yoo,E. S., and Kang, H. K. 2009. Apoptosis inducing activity of fucoidan in HCT-15 colon carcinoma cells. Biological and Pharmaceutical Bulletin 32 (10) 1760-1764.
| Hwang, P. A., Chien,S. Y., Chan,Y. L., Lu, M. K., Wu, C. H., Kong, Z. L., and Wu, C. J. 2011. Inhibition of lipopolysaccharide (LPS)- induced inflammatory responsesby Sargassum hemiphyllum sulfated polysaccharide extract in RAW264.7 macrophage cells. Journal of Agricultural and Food Chemistry 59: 2062-2068.
| Ito, K., and Hori, K. 1989. Seaweed: Chemical composition and  otential food uses. Food Reviews International 5: 101-144.
| Itoh, H., Noda, H., and Amano, H.  1993.  Antitumor activity and immunological properties of marine algal polysaccharide, especially fucoidan, prepared from Sargassum thunbergii of phaeophycease.  Anticaner Research  13(6A): 2045-2052.
| Kazlowski, B.,Chiu Y. H., Kazlowska, K., Pan, C. H. Wu, C. J. 2012. Prevention of Japanese encephalitis virus infections by low-degree-polymerisation sulfated saccharides from Gracilaria sp. and Monostroma nitidum. Food Chemistry 133: 866-874.
| Kazlowski, B., Ko,Y. T., and Pan, C. L. 2007. Comparison of two neoagaro-oligosaccharide production systems in the quantification of their oligomer products generated from agarose by -agarase digestion.  December 7th, 2007. B-56. Taiwan Association for Food Science and Technology, Changhua, Taiwan, R.O.C.
| Kazlowski, B., and Pan, C. L.  2008.  Separation and quantification of Monostroma-oligosaccharide from Monostroma nitidum and neoagaro-oligosaccharide from Gracilaria tenuistipitata products obtained via digestion of Aeromonas salmonicida MAEF108 -agarases by preparative HPLC and NMR analysis.  June 30th, 2008.  C42, p. 125.  Agricultural Chemical Society of Taiwan, Taipei, Taiwan, R.O.C.
| Kazlowski, B., Pan, C. L., and Ko, Y. T. 2008a. Separation and quantiﬁcation of neoagaro- and agaro-oligosaccharide products generated from agarose digestion by -agarase and HCl in liquid chromatography systems. Carbohydrate Research  343: 2443-2450.
| Kazlowski, B., Kazlowska, K., Pan, C. L., and Tsai, G. J. 2008b.  Stimulation on nitric oxide production in RAW 264.7 macrophages by defined chain lengths oligosaccharides derived from Gracilaria tenuistipitata and Monostroma nitidum.  May 8th to 9th, 2008.  BT-03, p. 270.  International Conference of Advanced Research on Marine Bioresources, Keelung, Taiwan, R.O.C.
| Kleerebezem, M., Beerthuyzen, M. M., Vaughan, E. E., Devos, W. M., and Kuipers, O. P. 1997. Controlled gene expression systems for lactic acid bacteria: Transferable nisin-inducible expression cassettes for Lactococcus, Leuconostoc, and Lactobacillus spp. Applied and Environmental Microbiology  63: 4581-4584.
| Kuipers, O. P., de Ruyter, P. G. G. A., Kleerebezem, M., and de Vos, W. M. 1998. Quorum sensing-controlled gene expression in lactic acid bacteria.  The Journal of  Bacteriology  64: 15-21.
| Laitinen, L., Kangas, H., Kaukonen, A. M., Hakala, K., Kotiaho, T., Kostiainen, R., and Hirvonen, J. 2003. N-in-one permeability studies of heterogeneous sets of compounds across Caco-2 cell monolayers. Pharmaceutical Research 20: 187-197.
| Lapikova, E. S., Drozd, N. N., Tolstenkov, A. S., Makarov, V. A., Zvyagintseva, T. N., Shevchenko, N. M., Bakunina, I. U., Besednova, N. N., and Kuznetsova, T. A. 2008. Inhibition of thrombin and factor Xa by Fucus evanescens fucoidan and its modified analogs. Bulletin of Experimental Biology and Medicine 46(3): 328-333.
| Lee, J. B., Hayashi, K., Hashimoto, M., Nakano, T., and Hayashi, T. 2004. Novel antiviral fucoidan from sporophyll of Undaria pinnatifida (Mekabu). Chemical and Pharmaceutical Bulletin 52: 1091-1094.
| Li, B., Lu, F., Wei, X., and Zhao, R. 2008. Fucoidan: Structure and bioactivity. Molecules 13: 1671-1695.
| Li, B., Wei, X. J., Sun, J. L., and Xu, S. Y. 2006. Structural investigation of a fucoidan containing a fucose-free core from the brown seaweed, Hizikia fusiforme. Carbohydrate Research 341: 1135-1146.
| Lowry, O. H., Rosebrough, N. J., Farr, A. L., and Randall, R. J. 1951 Protein measurement with the folin phenol reagent. Journal of Biological Chemistry 193: 265-275.
| Mandal, P., Mateu, C. G., Chattopadhyay, K., Pujol, C. A., Damonte, E. B.,  Ray, B. 2007. Structural features and antiviral activity of sulphated fucans from the brown seaweed Cystoseira indica. Antiviral Chemistry and Chemotherapy 18: 153-162.
| Maria, E. R., Cardoso, D. M. A., Noseda, M. D., Cerezo, A. S. 2001. Structural studies on fucoidans from the brown seaweed Sargassum stenophyllum. Carbohydrate Research 333:  281-293.
| Marinho-Soriano, E., Fonseca, P. C., Carneiro, M. A. A., and Moreira,  W. S. C. 2006. Seasonal variation in the chemical composition of two tropical seaweeds. BioresourceTechnology. 97: 2402-2406.
| Matsubara, K., Y. Mastsuura, A. Bacic, M.-L. Liao, K. Hori, and K. Miyazawa. 2001. Anticoagulant properties of a sulfate galactan preparation from a marin green alga, Codium cylindricum.  Biomacromolecules  28: 395-399.
| McKay, L. L., Balwin, K. A., and Zottola, E. 1972. Loss of lactose metabolism in lactic streptococci.  Applied Microbiology  23: 1090-1096.
| Mierau, I., and Kleerebezem, M. 2005. 10 years of the nisin-controlled gene expression system (NICE) in Lactococcus lactis.  Applied Microbiology and Biotechnology  9: 1-13.
| Mierau, I., Leij, P., van Swam, I., Blommestein, B., Floris, E., Mond, J., and Smid, E. J. 2005. Industrial-scale production and purification of a heterologous protein in Lactococcus lactis using the nisin-controlled gene expression system NICE®: The case of lysostaphin. Microbial Cell Factories 4: 15-24.
| Miller, G. L. 1959. Use of dinitrosalicylic acid reagent for reagent for deterimination of reduceing sugars. Analytical Chemistry 31: 426-428.
| Moll, S., and Roberts, H. R. 2002. Overview of anticoagulant drugs for the future.  Seminars in Hematology 39 ( 3): 145-157.
| Nagai, T., and Yukimoto, T. 2003. Preparation and functional properties of beverages made from sea algae.  Food Chemistry 81: 327-332.
| Nishide, E.  1981.  Extraction of fructose containing polysaccharide from the brown alga Kjellmanidlla crassfolia.  Bulletin of the Japanese Society of Scientific Fisheries  47: 1233-1235.
| Okai, Y., Higashi-Okai, H., Ishizaka, S., and Yamashita, U. 1997.  Enhancing effect of polysaccharides from an edible brown alga, Hijikia fusiforme (Hijihi), on release of tumor necrosis factor-alpha from macrophages of endotoxin-nonresponder C3H/HeJ mice.  Nutrition and Cancer 27: 74-79.
| Okai, Y., Higashi-Okai, K., Ishizaka, S., Ohtoni, K., Matsui-Yuasa, I., and Yamashita, U. 1998. Possible immunodulating activities in an extract of edible brown alga, Hijikia fusiforme (Hijihi). Journal of the Science of Food and Agriculture  76: 56-62.
| O’Sullivan, A. M., O’Callaghan Y. C., O’Grady, M. N., Queguineur, B., Hanniffy, D., Troy, D. J., Kerry, J. P., O’Brien, N. M. 2011. In vitro and cellular antioxidant activities of seaweed extracts prepared from five brown seaweeds harvested in spring from the west coast of Ireland.  Food Chemistry  126: 1064-1070.
| Oyaizu, M. 1986. Studies on products of the browning reaction. Antioxidative activities of browning reaction products prepared from glucosamine. Japanese Journal of Nutrition 44 (6): 307-315.
| Palkama, T. 1991.  Induction of interleukin-1 production by ligands binding to the scacenger recepter in human monocytes and the THP-1 cell line. Immunology 74: 432-438.
| Panesar, P., Kennedy, J. F., Gandhi, D. N., and Bunko, K. 2007.  Bioutilisation of whey for lactic acid production. Food Chemistry  105: 1-14.
| Pelegrin, Y. F., Robledo, D. R., and Garcia, G. 1995. Seasonal agar yield and quality in Gelidium canarieusis (Grunow) seoane-camba (Gelidiales, Rhodophyta) from gran canaria, Spain. The Journal of Applied Phycology 7: 141-144.
| Peterson, M. D., and Mooseker, M. S. 1992. Characterization of the enterocyte-like brush border cytoskeieton of the C2BBe clones of the human intestinal cell line, Caco-2. Journal of Cell Science 102: 581-600.
| Percival, E. and McDowell, R. H. 1967. Chemistry and Enzymology of Marine Algae Polysaccharide. Vol. 2, pp. 26-50. Academic Press.  New York, U.S.A.
| Pinto, M., Robine-Leon, S., Appay, M. D., Kedinger,M., Triadou, N., Dussaulx, E., Lacroix, B., Simon-Assmann, P., Haffen, K., Fogh, J., and Zweibaum, A. 1983. Enterocyte-like differentiation and polarization of the human colon carcinoma cell line Caco-2 in culture. Biology of the Cell 47: 323-330.
| Platteeuw, C., van Alen-Boerrigter, I., van Schalkwijk, S., and de Vos, W. M. 1996. Food-grade cloning and expression system for Lactococcus lactis.  Applied and Environmental Microbiology 62: 1008-1013.
| Plumb, J. A., Milroy, R., and Kaye, S. B. 1989 Effects of the pH- dependence of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl- tetrazolium bromide formazan absorption on chemosensitivity determined by a novel tetrazolium-based assay. Cancer Research 49: 4435-4440.
| Ponce, N. M. A., Pujol, C. A., and Damonte, E. B. 2003. Fucoidans from the brown seaweed Adenocystis utricularis: Extraction methods, antiviral activity and structural studies. Carbohydrate Research 338: 153-165.
| Preetha, S. P., and Devaraj, H. 2010. Role of sulphated polysaccharides from Sargassum wightii in the control of diet-induced hyperlipidemia and associated inflammatory complications in rats. European Journal of  Inflammation 8 23-30.
| Robledo, D., and Freile-Pelegrin, Y. 1997. Chemical and mineral omposition of six potentially edible seaweed species of Yucata’n. Botanica Marina 40: 301-306.
| Sambrook, J., and Russell, D. W.  2006.  The Condensed Protocol from Molecular Cloning: A Laboratory Manual.  Cold Spring Harbor Lab. Press, Cold Spring Harbor, NY, U.S.A.  pp. 58-59, 177, 188-192, 243, 718-725, 730, 758-760.
| Saunders, M. 2009. Transplacental transport of nanomaterials. WIREs Nanomedicine and Nanobiotechnology  1: 671-684.
| Shan, B. E., Yoshida, Y., Kuroda, E., and Yamashita, U. 1999.  Immunomodulating activity of seaweed extract on human lymphocytes in vitro. International Journal of Immunopharmacology  21: 59-70.
| Shimada, K., Fujikawa, K., Yahara, K., and Nakamura, T. 1992. Antioxidative properties of xanthan on the antioxidation of soybean oil in cyclodextrin emulsion. Journal of Agricultural and Food Chemistry 40: 945-948.
| Singleton, V. L., and Rossi, J. A. 1965. Colorimetry of total phenolics with phosphomolybdic-phosphotingsticacid regent. American Journal of Enology Viticulture 16:144-153.
| Sogawa, K., Sumida, T., Hamakawa, H., Yamada, T., Matsumoto, K., and Matsuda, H.  1998.  Inhibitory effect of a marine microalgal polysaccharide on telomerase activity in K562 cells.  Research Communications in Molecular Pathology and Pharmacology  9: 259-265.
| Stiles, M. E., and Holzapfel, W. H.  1997.  Lactic acid bacteria of foods and their current taxonomy.  International Journal of Food Microbiology  36: 1-29.
| Taoda, N., Shinji, E., Nishii, K., Nishioka, S.,  Yonezawa, Y.,  Uematsu,  J., Hattori, E., Yamamoto, H.,  Kawano, M., Tsurudome, M., O'Brien, M.,  Yamashita, T., and Komada, H. 2008. Fucoidan inhibits parainfluenza virus type 2 infection to LLCMK2 cells.  Biomedical Research  29: 331-334.
| Terho, T. T., and Hartiala, K. 1971.  Method for determination of the sulfate content of glycosaminoglycans. Analytical Biochemistry 41: 471-476.
| Terzaghi, B. E., and Sandine W. E. 1975. Improved medium for lactic streptococci and their bacteriophage.  Applied Microbiology 29: 807-813.
| Tavelin, S., Milovic, V., Ocklind, G., Olsson S., and Artursson, P. 1999. A conditionally immortalized epithelial cell line for studies of intestinal drug transport. Journal of Pharmacology and Experimental Therapeutics 290: 1212.
| Tavelin, S., Taipalensuu, J., Erberg, L. S., Morrison, R., Chong, S., and Artursson, P. 2003. Prediction of the oral absorption of low-permeability drugs using small intestine-like 2/4/A1 cell monolayers. Pharmaceutical Research 20: 397-405.
| Turvey, J. R., and William, E. L. 1970. The structure of some xylans from red algal.  Phytochemistry  9: 2383-2388.
| van den Ent, F., and Lowe, J. 2006. RF cloning: A restriction-free method for inserting target genes into plasmids. Journal of Biochemical and  Biophysical Methods  67: 67-74.
| Vincent, M. L., Russell, R. M., and Sasak, V. 1985. Folic acid uptake characteristics of a human colon carcinoma cell line, Caco-2. A newly-described cellular model for small intestinal epithelium. Human Nutrition: Clinical Nutrition 39: 355-360.
| Walgren, R. A, Walle, U. K, and Walle, T. 1998. Transport of Quercetin and Its Glucosides across Human Intestinal Epithelial Caco-2 Cells. Biochemical Pharmacology 55: 1721-1727.
| Wang, Y., Yu, G. L., Zhao, X., Guan, H. S., and Du, Y. G. 2005. Preparation and structural characterization of several oligoguluronic acids. Chemical Journal of Chinese Universities  26: 179-183.
| Wang, J., Zhang, Q., Zhang, Z., Song, H., and Li, P. 2010. Potential antioxidant and anticoagulant capacity of low molecular weight fucoidan fractions extracted from Laminaria japonica. International Journal of Biological Macromolecules  46: 6-12.
| Wang, J., Zhang, Q., Zhang, Z., Song, H., and Zhien, L. 2008. Antioxidant activity of sulfated polysaccharide fractions extracted from Laminaria japonica. International Journal of Biological Macromolecules  42: 127-132.
| Weerachayaphorn J., and Pajor A. M. 2008. Identification of transport pathways for citric acid cycle intermediates in the human colon carcinoma cell line, Caco-2. Biochimica et Biophysica Acta (BBA) - Biomembranes 1778(4): 1051-1059.
| Wells, J. M., and Le-Page, R. W. F.  1993. Improved cloning vectors and transformation procedure for Lactococcus lactis.  Journal of Applied Bacteriology  74: 629-636.
| Whistler, R. L. 1965. Methods in Carbohydrate Chemistry. Vol. 5, pp. 3-46. Academic Press. New York, U.S.A.
| Wisselink, H. W., Weusthuis, R. A., Eggink, G., Hugenholtz, J., and Grobben, G. J. 2002. Mannitol production by lactic acid bacteria: A review.  International Dairy Journal 12: 151-161.
| Xue, C. H., Yu, G. L., Hirata, T., Terao, J., and Lin, H. 1998. Antioxidative activities of several marine polysaccharides evaluated in a phosphatidylcholine-liposomal suspension and organic solvents. Bioscience, Biotechnology, and Biochemistry 62: 206-209.
| Yamasaki-Miyamoto, Y., Yamasaki, M., Tachibana, H., and Yamada, K. 2009. Fucoidan induces apoptosis through activation of caspase-8 on human breast cancer MCF-7 cells. Journal of Agricultural and Food Chemistry 57: 8677-8682.
| Yang, H. N. 2010. Biota Taiwanica. http://algae.biota.biodiv.tw/pages/16
| 24 (June 25, 2012, Taiwan)
| Yu, H., Cook, T. J., Sinko, P. J. 1997. Evidence for diminished functional expression of intestinal transporters in Caco-2 cell monolayers at high passages. Pharmaceutical Research 14: 757-762.
| Yu, P., Li, N., Liu, X., and Zhou, G. 2003. Antiperlipidemic effects of different molecular weight sulfated polysaccharides from Ulva pertusa (Chlorophyta).  Pharmacological Research  48: 543-549.
| Yuan, H., Song, J., Li, X., Li, N., and Dai, J. 2006.  Immunomodulation and antitumor activity of -carrageenan oligosaccharides. Cancer Letter 243: 228-234.
| Zhang, Z., Zhang, Q., Wang, J., Song, H.,  Zhang, H., and Niu, X.2010. Regioselective syntheses of sulfated porphyrans from Porphyra haitanensis and their antioxidant and anticoagulant activities in vitro. Carbohydrate Polymers 79: 1124-1129.
| Zhao, X., Xue, C., Cai, Y., Wang, D., and Fang, Y. 2005. Study of antioxidant activities of fucoidan from Laminaria japonica. High Technology Letters 11: 91-94.
| Zhu, Z., Zhang, Q., Chen, L., Ren, S., Xu, P., Tang, Y., and Luo, D. 2010. Higher specificity of the activity of low molecular weight fucoidan for thrombin-induced platelet aggregation. Thrombosis Research 125: 419-426.
|

|

預覽及輸出檢索結果

記錄

欄位

| 本頁 1 筆
| 記錄 1 至 50
| 選擇之記錄

| 標題
| 全部欄位
| selected fields new window
| APA Style
| Chicago Style
| MLA Style
| CNS-13611 Style
| EndNote Style

e-mail 地址 | : |
e-mail 標題 | : |

| |

系統規劃：海洋大學圖書館
系統製作：飛資得資訊有限公司

2012 年 11 月 19 日 8:55