To Study the Production of Seaweed Oligosacchrides by High Hydrostatic Pressure and to Evaluate Its Bioactivities(2/2)

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Project title

Code/計畫編號

MOST108-2221-E019-043-MY2

Translated Name/計畫中文名

探討以高靜水壓生產海藻寡醣並評估其生物活性(2/2)

Project Coordinator/計畫主持人

Chun-Ju Chang

Funding Organization/主管機關

National Science and Technology Council

Department/Unit

Department of Food Science

Website

https://www.grb.gov.tw/search/planDetail?id=13327724

Year

2020

Start date/計畫起

01-08-2020

Expected Completion/計畫迄

31-07-2021

Bugetid/研究經費

1201千元

ResearchField/研究領域

食品科技(工)

"研究背景：我們過去以海洋細菌酵素生產的低分子量海藻多醣證實具有益生性，並具有抑制雙醣酶、抗氧化、抗凝血等功能，但以酵素生產海藻多醣之成本高、產率低，因此本研究目標之一為改良海藻寡醣生產方式。另一方面，國內外目前均無海藻寡醣之腸道吸收運輸途徑相關證據，本研究目標之二為探討海藻寡醣於腸道吸收之作用機制。研究方法：購買無汙染海水養殖之食用綠藻臺灣小葉蕨藻 (Caulerpa lentillifera) 為原料，以高靜水壓 (high hydrostatic pressure，HHP) 破壞藻體細胞壁以提升多醣產率，探討原料用量、壓力及加工時間等三項變因對藻多醣產率之影響，並以反應曲面法（response surface methodology）預測最適生產條件。再以超過濾系統（ultrafiltration）收集分子量低於1 kDa之海藻寡醣 (seaweed oligosaccharides, SO)，分析其分子量、預測聚合度，並評估SO之抗氧化、抗腫瘤及降血糖、降血壓、降血脂等生物活性。另一方面以螢光標定SO追蹤其在人類腸道屏障Caco-2細胞單層之吸收運輸率；應用免疫螢光染色分別標記腸道單層的肌動蛋白F-actin、緊密連結運輸指標ZO-1或內吞作用蛋白Rab5/Rab11，以共軛焦顯微鏡觀察SO在腸道運輸過程之分布變化；進一步以抑制劑驗證以上運輸途徑在海藻寡醣吸收過程之關鍵角色。預期結果：本研究第一年將建立以HHP生產海藻寡醣之最適化條件並評估產物之保健潛力。第二年將呈現海藻寡醣在腸道吸收運輸的主要途徑、探討tight junction及內吞作用(endocytosis) 相關蛋白在海藻寡醣運輸過程的重要性。" "Background: Our previous studies documented that marine bacterial enzymes produced low molecular weight seaweed oligosaccharides (SO) have anti-disaccharidases, antioxidant and anti-coagulation. However, high cost of enzymes and low yield of SO limited further investigation. Consequently, aim 1 of this project is to modify the process of functional SO. Besides, the intestinal absorption and transport pathways of SO are unclear. Therefore, aim 2 of this project is to investigate the intestinal absorption mechanisms of SO. Methods: Cultivate Caulerpa lentillifera will be purchased from Taiwan sea-farmer. Cell walls of seaweed will be broke down by high hydrostatic pressure (HHP). Meanwhile, the optimal conditions for HHP will be analyzed by response surface methodology. SO (molecular weight less than 1 kDa) will be isolated by ultrafiltration. The molecular profile and degree of polymerization of SO will be analyzed by HPLC. The antioxidant, anti-cancer, and hypoglycemic, hypotensive as well as hypolipidemic properties of SO will be evaluated. On the other hand, SO will be labelled by fluorescein then the absorption and transport rate of SO will be measured in human intestinal Caco-2 monolayer. By immunofluorescent stain, the distribution of SO in Caco-2 monolayer, as well as F-actin, marker of tight junction ZO-1, and endocytosis protein Rab5/Rab11 will be visualized by confocal microscopy. Furthermore, the importance of these intestinal transport pathways for SO will be characterized by pretreatment of transport inhibitors. Anticipated results: In the first year, this project will demonstrate the optimized condition for HHP, and prove the health benefit of SO. In the second year, the intestinal absorption pathway of SO will be revealed. The roles of tight junction, and endocytosis-related proteins on transportation of SO will be established."

Keyword(s)

高靜水壓
反應曲面法
腸道Caco-2細胞單層
內吞作用
high hydrostatic pressure
response surface methodology
intestinal Caco-2 monolayer
endocytosis

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To Study the Production of Seaweed Oligosacchrides by High Hydrostatic Pressure and to Evaluate Its Bioactivities(2/2)

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