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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 吳瑞碧(James Swi-Bea Wu) | |
dc.contributor.author | Chih-Fang Lin | en |
dc.contributor.author | 林志芳 | zh_TW |
dc.date.accessioned | 2021-06-15T00:33:29Z | - |
dc.date.available | 2009-01-20 | |
dc.date.copyright | 2009-01-20 | |
dc.date.issued | 2008 | |
dc.date.submitted | 2009-01-12 | |
dc.identifier.citation | 莊榮輝。2005。酵素化學。國立台灣大學 農業化學系生物化學研究室
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dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/41830 | - |
dc.description.abstract | 微生物轉麩胺酸醯胺基酶(Microbial transglutaminase; MTGase)催化蛋白質分子間或分子內形成ε-(γ-glutamyl)lysine共價鍵,可用來轉變分子結構以改善食品中蛋白質性質。本研究直接採用市售MTGase粗酵素粉末,經過加水萃取,此粗酵素液先用50~80%硫酸銨沈澱,可得最大酵素活性;再經過CM-Sepharose CL-6B膠濾層析,以及Blue Sepharose 6 Fast Flow 親和性管柱分離,經過這幾個步驟,以SDS-PAGE證明,可得到純化的MTGase。
米穀粉添加不同比例MTGase,以動態流變儀(Rheometer)測其黏彈性,顯示出其彈性模式(G′)與黏性模式(G〞)均隨MTGase添加而增加;至於質地組織分析(texture analysis),其質地參數(硬度、膠性)也是隨MTGase添加而增加;在熱性質測試,添加MTGase因其作用蛋白質分子,交叉鍵結形成聚合分子,有降低焓值的現象(enthalpy)。但值得注意的是添加過多的MTGase(0.3U/mg,以米蛋白質為基重)反而降低其膠體強度與TPA各種物性,此可能會由於太多ε-(γ-glutamyl)lysine的鍵結會抑制蛋白質網狀結構的發展而降低其彈性,以添加MTGase 0.1U/mg與0.2U/mg rice protein可得較佳效果,所以此為建議添加量。SDS-PAGE profile證明MTGase會鍵結米蛋白質形成較大分子量的聚合物(HMW)。由電子顯微掃瞄(SEM)可觀察到,MTGase添加量愈多,米中蛋白質結構愈細緻,孔洞愈小,而且排列愈呈組織化。 以另一種方式探討MTGase影響秈米中蛋白質學性質,將秈米的蛋白質依不同溶劑中溶解性不同來區分,其萃出可分為四大類:水溶(albumin)、鹽溶(globulin)、鹼溶(glutelin)以及醇溶(prolamin),含量多寡依序為鹼溶蛋白>鹽溶蛋白>水溶蛋白>醇溶蛋白。醇溶蛋白,經SDS-PAGE分析只有一個次單位(subunit)。 從動態流變性質測知,白米中水溶蛋白(albumin)對增進河粉膠體強度較無貢獻,所以建議米食加工時可以去除。米蛋白中,穀蛋白的半胱胺酸含量多,對產品的影響較大,由SDS-PAGE圖可觀察到添加MTGase的交聯效果。 | zh_TW |
dc.description.abstract | Microbial transglutaminase (MTGase) may catalyze the cross-linking between a peptide-bound glutaminyl residue and an ε-amino group of lysine residue in protein. MTGase has been used to modify food protein for improving their functionality. Not pure MTGase could be supplied, so making it by myself. The commercial microbial transglutaminase was obtained from Ajinomoto Co.(Japan). Added to water and followed by centrifugation. The supernatant was filtered to obtain the crude enzyme solution. The solution was precipitated with ammonium sulfate. In the obtained precipitates, 50~80% showed the better enzyme activity. Then the supernatant was purified by gel filtration through a CM- Sepharose CL-6B column and a Blue Sepharose Fast Flow column. Used SDS-PAGE profile could prove to get pure MTGase.
Rice flour was treated with different concentrations of MTGase. The rheological properties, elastic (G') and viscous (G') moduli of the rice gel increased at the addition of MTGase. However, high amount of the addition of MTGase at 0.3U/mg may reduce textural parameters. Therefore, rheological properties and textural parameters might be affected by the existing threshold number of connections for cross-links in rice protein. Differential scanning calorimetry (DSC) showed that MTGase treatment led to decrease the denaturation enthalpy. Sodium dodecyl sulfate- polyacrylamide gel electrophoresis (SDS-PAGE) confirmed that rice proteins were polymerized through the MTGase reaction. Scanning electron microscope (SEM) figure showed the more concentrations of MTGase added to rice flour, the better texture of rice flour gel. Therefore, those SEM photographs proved the added MTGase improved the rheological properties and textural parameters (hardness, and gumminess). Here another method was used to investigate the effects of MTGase to properties of rice protein. Rice protein was treated with different solvents and there were four kinds of extractions were obtained: water soluble (albumin), salt soluble (globulin), alkali soluble (glutelin) and alcohol soluble (prolamin) fractions. The amount obtained were in the order of alkali soluble protein>salt soluble protein>water soluble protein>alcohol soluble protein. After SDS-PAGE analysis, alcohol soluble protein only one subunit. From Rheology test, water soluble protein (albumin) was proved has no contribution in strength of rice gel. Therefore it is suggested to remove it at the rice processing. Rice protein contain much cysteine, and affects the properties of the products. The rice contains much glutelin, from SDS-PAGE profile, the effects of MTGase addition to the linkages were observed. | en |
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dc.description.tableofcontents | 中文總摘要……………………………………………………………………… Ⅰ
英文總摘要……………………………………………………………………… Ⅲ 目錄……………………………………………………………………………… Ⅴ 圖目錄…………………………………………………………………………… Ⅹ 表目錄…………………………………………………………………………… ⅩⅡ Ⅰ、微生物轉麩胺酸醯胺基酶純化……………………………………… 1 中文摘要……………………………………………………………………… 2 英文摘要……………………………………………………………………… 3 前言……………………………………………………………………………… 4 文獻整理………………………………………………………………………… 6 壹、轉麩胺酸醯胺基酶特性及其應用……………………………………… 6 一、轉麩胺酸醯胺基酶性質……………………………………………… 6 二、轉麩胺酸醯胺基酶來源……………………………………………… 6 三、TGase的分子特性…………………………………………………… 8 四、Microbial Transglutaminase分子性質………………………………… 10 五、TGase的活性測定…………………………………………………… 10 六、終止TGase活性測定反應…………………………………………… 11 七、影響TGase作用因子………………………………………………… 12 貳、麩胺醯胺轉胺酶之應用………………………………………………… 13 一、MTGase對凝膠作用之影響…………………………………… 13 二、MTGase對煉製品之影響……………………………………… 13 三、MTGase對肉製品之應用……………………………………… 14 四、MTGase對乳化作用之影響………………………………… 16 五、MTGase在小麥產品之應用…………………………………… 16 六、MTGase在其他方面之應用…………………………………… 17 參、MTGase之安全性…………………………………………………… 19 材料與方法……………………………………………………………………… 20 壹、實驗材料………………………………………………………………… 20 貳、儀器設備………………………………………………………………… 21 參、實驗方法………………………………………………………………… 22 一、蛋白質含量測定………………………………………………… 22 二、硫酸銨劃分………………………………………………………… 23 三、MTGase活性測定……………………………………………… 23 四、聚丙烯醯胺膠體電泳(SDS-PAGE)分析…………………… 27 五、實驗步驟…………………………………………………………… 30 結果與討論……………………………………………………………………… 32 一、MTGase萃取……………………………………………………… 32 二、MTGase硫酸銨劃分…………………………………………… 33 三、CM Sepharose CL-6B 離子交換層析………………………………… 35 四、Blue Sepharose Fast Flow管柱層析………………………………… 37 五、MTGase SDS-PAGE圖……………………………………………… 39 結論…………………………………………………………………………… 42 參考資料……………………………………………………………………… 43 Ⅱ、微生物轉麩胺酸醯胺基酶對河粉膠體性質之影響………… 53 中文摘要……………………………………………………………………… 54 英文摘要……………………………………………………………………… 55 前言…………………………………………………………………………… 57 文獻整理……………………………………………………………………… 59 壹、米粒結構與特性……………………………………………………… 59 一、米粒結構…………………………………………………………… 59 二、米穀粉……………………………………………………………… 60 貳、影響米穀粉理化性質之因素…………………………………… 60 一、米粒結構…………………………………………………………… 60 二、米蛋白對米穀粉糊化性質之影響…………………………… 61 三、米種選擇…………………………………………………………… 62 參、食品的流變學………………………………………………………… 62 一、流變學定義………………………………………………………… 62 二、流體的種類………………………………………………………… 62 三、流變儀(rheometer)原理……………………………………… 64 四、流變性質…………………………………………………………… 65 五、流變性質在食品的應用………………………………………… 66 材料與方法…………………………………………………………………… 67 壹、實驗材料………………………………………………………………… 67 貳、儀器設備………………………………………………………………… 67 參、實驗方法………………………………………………………………… 68 一、一般成分分析…………………………………………… 68 二、蛋白質含量測定………………………………………… 68 三、蛋白質萃出液…………………………………………… 68 四、製作河粉膠體…………………………………………… 69 五、流變性質………………………………………………… 70 六、河粉膠體的質地分析…………………………………… 70 七、示差熱掃瞄分析………………………………………… 71 八、米蛋白之電泳分析……………………………………… 71 九、掃瞄式電子顯微鏡……………………………………… 72 十、數據之統計分析………………………………………… 72 實驗架構………………………………………………………………………… 73 一、製備米穀粉……………………………………………………… 73 二、實驗流程………………………………………………………… 73 結果與討論……………………………………………………………………… 76 壹、米穀粉與MTGase作用的流變性質…………………………… 76 貳、微生物轉麩胺酸醯胺基酶對河粉膠體質地之影響……… 80 參、MTGase對米穀粉熱性質之影響……………………………… 83 肆、米蛋白聚丙烯醯胺膠體電泳(SDS-PAGE)分析…………… 88 伍、電子顯微掃瞄……………………………………………………… 92 結論……………………………………………………………………………… 100 參考文獻………………………………………………………………………… 101 Ⅲ、米中不同區分蛋白質與MTGase對穀蛋白之影響……………… 109 中文摘要……………………………………………………………………… 110 英文摘要……………………………………………………………………… 111 前言…………………………………………………………………………… 113 文獻整理……………………………………………………………………… 114 米中蛋白質的種類與特性……………………………………………… 114 一、米中蛋白質的種類……………………………………………… 114 二、米中水溶蛋白質的特性…………………………………… 114 三、米中鹽溶蛋白質的特性………………………………………… 115 四、米中鹼溶蛋白質的特性………………………………………… 115 五、米中醇溶蛋白質的特性………………………………………… 115 材料與方法……………………………………………………………………… 117 壹、實驗材料………………………………………………………………… 117 貳、儀器設備………………………………………………………………… 117 參、實驗方法………………………………………………………………… 118 一、米蛋白萃取區分………………………………………………… 118 二、實驗流程…………………………………………………………… 119 三、製作去除白蛋白之河粉膠體……………………………… 120 四、米中穀蛋白(glutelin)與MTGase反應…………………… 121 結果與討論……………………………………………………………………… 122 壹、米蛋白質性質………………………………………………………… 122 一、秈米中成分………………………………………………………… 122 二、米蛋白萃取………………………………………………………… 123 三、米蛋白性質探討………………………………………………… 125 貳、添加MTGase對去除白蛋白的河粉膠體之影響…………… 128 參、米中穀蛋白與MTGase反應……………………………………… 132 結 論…………………………………………………………………………… 135 參考文獻………………………………………………………………………… 136 | |
dc.language.iso | zh-TW | |
dc.title | 轉麩胺酸醯胺基酶處理對河粉膠體物性影響之研究 | zh_TW |
dc.title | Modification in physical properties of rice gel by microbial transglutaminase | en |
dc.type | Thesis | |
dc.date.schoolyear | 97-1 | |
dc.description.degree | 博士 | |
dc.contributor.oralexamcommittee | 盧訓(Shin Lu),黃卓治(Tzou-Chi Huang),王裕泰(Yuh-Tai Wang),吳明昌(Ming-Chang Wu),江啟銘(Chii-Ming Jiang) | |
dc.subject.keyword | 微生物轉麩胺酸醯胺基酶,CM Sepharose CL-6,Blue Sepharose fast Flow親和性膠濾層析、流變性質、質地組織、聚丙烯醯胺膠體電泳,電子顯微掃瞄,白蛋白,球蛋白,穀蛋白,醇溶蛋白, | zh_TW |
dc.subject.keyword | Microbial transglutaminase (MTGase),Sepharose CL-6B,Blue Sepharose Fast Flow, rheology, TPA, DSC,SDS-PAGE,SEM,albumin,globulin,glutelin,prolamin, | en |
dc.relation.page | 140 | |
dc.rights.note | 有償授權 | |
dc.date.accepted | 2009-01-12 | |
dc.contributor.author-college | 生物資源暨農學院 | zh_TW |
dc.contributor.author-dept | 食品科技研究所 | zh_TW |
顯示於系所單位: | 食品科技研究所 |
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