臺灣博碩士論文加值系統

膠原蛋白（collagen）是近年來相當受到重視的生醫材料，其約佔脊椎動物體總蛋白質的30％，主要分佈於結締組織，是構成結締組織的重要結構性蛋白質。在生理溶液中膠原蛋白會聚集形成纖維基質，成為結構性的骨架，在結締組織與肌肉或器官共司各種新陳代謝機能時，膠原蛋白即扮演支持、保護及有關水分、離子等吸附或輸送之角色。目前為止已發現了七大類共十九型之膠原蛋白，其中，第二型膠原蛋白被證明可用於治療類風濕性關節炎。口服第二型膠原蛋白可調節體內之免疫反應，避免軟骨組織受到T細胞之攻擊。第二型膠原蛋白主要分佈於軟骨之中，可見於雞胸軟骨、牛鼻軟骨、椎間盤及玻璃液等處。本省雞胸軟骨及雞腳年產約510及30,434公噸，除了食用，大多製成肉骨粉或廢棄，若能加以利用將可提高其經濟價值。本試驗即使用不同體積比之3M氯化鎂及2M氯化鈣溶液對雞胸軟骨及5％乳酸浸泡24小時明膠萃取後之雞腳骨渣進行第二型膠原蛋白之萃取。
結果顯示：不同體積比之3M氯化鎂及2M氯化鈣溶液對雞胸軟骨蛋白多醣之移除效率以體積比1：3最佳，體積比1：2次之。對5％乳酸浸泡24小時明膠萃取後之雞腳骨渣蛋白多醣之移除效率則以體積比1：3最佳，體積比2：1次之。由雞胸軟骨及5％乳酸浸泡24小時明膠萃取後之雞腳骨渣所萃取出之第二型膠原蛋白電泳圖，發現其純度相當高。雞胸軟骨第二型膠原蛋白產率，以體積比1：3之3M氯化鎂及2M氯化鈣溶液最高。5％乳酸浸泡24小時明膠萃取後之雞腳骨渣第二型膠原蛋白產率以3M氯化鎂及2M氯化鈣溶液單獨處理組最高。雞胸軟骨第二型膠原蛋白產品中鹽分含量，以體積比1：1之3M氯化鎂及2M氯化鈣溶液最高。5％乳酸浸泡24小時明膠萃取後之雞腳骨渣第二型膠原蛋白產品中鹽分含量，以2M氯化鈣溶液處理組最高。試製添加20μg第二型膠原蛋白之即食味噌湯，官能品評成績以添加15g味噌之組別最佳，且添加之膠原蛋白並不會影響其風味。

Collagen is an important biomedical material in recently. It is approximately 30 percent of total protein in vertebrate and mainly distributed in connective tissue as an important structural protein to constitute connective tissue. Collagen also plays the role of support, protection and water/ion adsorption and transportation on metabolism in connective tissue, muscle or organ. A total of 19 types collagens was found in the science and type II collagen had been evidenced to efficiently cure rheumatoid arthritis. Oral administration of type II collagen can regulate immune system to avoid T cells attack chondrocyte. Type II collagen was mainly distributed in cartilage and usually found in chicken sternal cartilage, bovine nasal cartilage, intervertebral disc and vitreous humor. Chicken sternal cartilage and feet are the major by- products in poultry meat industry in Taiwan and almost used to make meat-bone meal by rendering plant. In this study, we will utilize these raw material to extract type II collagen as an biomedical material to elevate their added-value. Different ratios of 3M MgCl2 and 2M CaCl2 solutions were designed to remove proteoglycan and extract type II collagen from chicken sternal cartilage and chicken feet residues soaking in 5% lactic acid for 24hrs.
The results showed that the best removal efficiency of chicken sternal cartilage proteoglycan for the ratio of 3M MgCl2 and 2M CaCl2 solution was 1:3(v/v), and then the second was 1:2(v/v). The best removal efficiency of proteoglycan in chicken feet residues soaking in 5% lactic acid for 24hrs for the ratio of 3M MgCl2 and 2M CaCl2 solution was also 1:3(v/v), and then the second was 2:1(v/v). The SDS-PAGE electrophotogram of type II collagen product extracted from chicken sternal cartilage and chicken feet residues soaking in 5% lactic acid for 24hrs evidenced that the purity of type II collagen was similar to commercial products(Sigma). Type II collagen from chicken sternal cartilage extracted by 1:3(v/v) of 3M MgCl2 and 2M CaCl2 solution had the highest yield. The highest yield of type II collagen from chicken feet residues soaking in 5% lactic acid for 24hrs was found by 2M CaCl2 solution. On the other hand, type II collagen product from chicken sternal cartilage extracted by 1:1(v/v) of 3M MgCl2 and 2M CaCl2 solution had the highest salt content. The salt content of the sample from chicken feet residues soaking in 5% lactic acid for 24hrs was found the highest value by 2M CaCl2 solution. However, the sensory panel of ready-to-eat miso soup with 20μg type II collagen and 15g miso paste showed that the best acceptance by the panelists and the flavor of miso soup wasn’t affected by type II collagen addition.

壹、中文摘要………………………………………………………………1
貳、前言……………………………………………………………………3
參、文獻檢討………………………………………………………………4
一、結締組織之功能與組成……………………………………………4
（一）結締組織的分佈與其功能性………………………………… 4
（二）結締組織之分類……………………………………………… 4
(三) 結締組織之組成……………………………………………… 5
二、軟骨之組成、特性與分類…………………………………………5
（一）軟骨的組成與特性…………………………………………… 5
(二) 軟骨的分類…………………………………………………… 6
三、軟骨的細胞外基質之組成…………………………………………7
四、蛋白多醣之組成、生理重要性與萃取方法………………………7
（一）蛋白多醣之組成……………………………………………… 7
（二）軟骨的蛋白多醣聚集體與其功能性………………………… 9
（三）蛋白多醣生理重要性………………………………………… 11
（四）蛋白多醣的移除方法………………………………………… 11
五、膠原蛋白之結構、生化合成、分類及組織分佈…………………12
（一）膠原蛋白之結構……………………………………………… 12
（二）膠原蛋白之生化合成………………………………………… 13
（三）膠原蛋白之分類及其在組織的分佈………………………… 18
六、膠原蛋白之應用……………………………………………………28
七、類風濕性關節炎（rheumatoid arthritis, RA）………………31
（一）類風濕性關節炎發生的原因……………………………………31
（二）類風濕性關節炎產生的症狀與診斷方式………………………31
（三）類風濕性關節炎的治療…………………………………………34
肆、材料與方法……………………………………………………………36
一、原料之來源……………………………………………………… 36
二、原料前處理……………………………………………………… 36
三、第二型膠原蛋白之萃取………………………………………… 37
（一）蛋白多醣之移除…………………………………………… 37
（二）第二型膠原蛋白之萃取…………………………………… 38
四、測定項目………………………………………………………… 39
（一）一般化學組成……………………………………………… 39
（二）己糖醛酸（hexuronic acid）分析……………………… 42
（三）第二型膠原蛋白之產率…………………………………… 44
(四) 食鹽含量測定……………………………………………… 44
（五）電泳分析…………………………………………………… 45
（六）即食產品之製造與官能品評……………………………… 50
（七）統計分析…………………………………………………… 51
伍、結果與討論……………………………………………………………52
一、原料之一般化學組成……………………………………………52
二、蛋白多醣移除效率………………………………………………54
三、第二型膠原蛋白產品……………………………………………58
四、第二型膠原蛋白之電泳圖………………………………………62
五、第二型膠原蛋白之產率…………………………………………65
六、第二型膠原蛋白產品之鹽分含量………………………………69
七、即食產品之外觀與官能品評……………………………………73
陸、結論……………………………………………………………………76
柒、參考文獻………………………………………………………………78
捌、英文摘要………………………………………………………………86

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