臺灣博碩士論文加值系統

植酸常被視為抗營養分子，在植物中為磷酸根儲存的主要形式；本研究使用含E. coli 植酸酶基因appA的轉殖水稻中，將轉殖基因稻米粉加入飼料，探討其對礦物質吸收的影響。本實驗除了測量市面上不同品牌之糙米、胚芽米和白米之植酸以及糙米不同時間發芽之植酸酶活性，還有將轉基因米粉直接加入米漿粉中，混合之後餵食老鼠，觀察其重量變化，及礦物質生物可利用性。
市面上不同品牌之稻米會經磨成粉後，測其植酸含量；糙米發芽是將糙米浸水，溫水洗滌後，靜置暗處，於不同時間測量其植酸酶之變化；動物實驗部分成四組，第一組為控制組，第二、三及四組於米漿粉飼料中加入一倍、兩倍及三倍可在30分鐘內將飼料中的植酸完全水解之轉基因米粉量。將飼料餵食24隻wistar大鼠，每組6隻，餵食28天，每天記錄攝食情況且收集糞便，每星期檢測動物重量和糞便重量。將飼料和收集來的糞便磨粉經溼式灰化後，用石墨原子吸收光譜分析飼料及糞便中的礦物質鈣、鋅、鐵及鎂的含量，用呈色法測量磷的含量。
結果發現糙米以中興牌之植酸較高，胚芽米是金墩牌之植酸較高，白米是台糖牌之植酸較高，糙米發芽所產生之植酸酶是隨著發芽時間而增加，動物實驗部分有添加轉基因米粉的組別，其鈣、鐵、鋅、磷及鎂生物可利用性皆有明顯的改善；其中鈣的吸收率最高，其次鋅、鐵及鎂。糞便中植酸降低率是添加3倍轉殖基因米粉的效果較佳，約可降低10%的植酸殘留。

ice milk flavored with roasted peanut is a popular breakfast beverage to Chinese. Borh rice and peanut contain phytate which is considered to be an anti-nutritional factor to the absorption of metals , protein and starch . A limited phytase, an enzyme specific for hydrolysis of phytate , has been reported to occur in the grain and oilseed. Including into food / feed a stable and high activity phytase will improve the absorption of nutrients , especial metal ions. In this study we will examine the phytate content of rice milk components , rice and peanut, and phytase activity during germination first. Then we will include a stable, high activith of phytase from transgenic rice to feed for mice to investigate its effect on the absortion of metals.
The contents of phytate in different brands of brown rice , embroyed rice and polished rice from market have been investigated. It was found that generally the content of phytate in rice is in following order: brown rice> embroyed rice >> polished rice for same brand. Phytate content in 100g of Sunhow rice were found to be 1.24, 1.23 and 0.07 g for brown rice, embroyed rice and polished rice, respectively. Phytase activity in germinated rice is increased with increase in germination duration. However, phytase activity of germinated regular rice (<1200U/kg) is much lower than that of the transgenic rice (30000-40000 U/kg).
Twenty four Wistars male divided into four groups. Control group was fed basal diet containing regular rice powder. In group 2, the calculated amount of transgenic rice required to hydrolyze total phytate in feed in 30 minutes was incorporated into basal diet to replaced partial regular rice. Double and triple transgenic rice were added into basal diet in group 2 and 3, respectively. Each group was fed for 4 weeks. The feed consumption and collected feces were recorded daily. The body weights were measured weekly. The diet and collected feces were dried, powdered and ashed for measurement of calcium, zinc, iron and magnesium content with graphite atomic absorption spectrophotometer and phosphorous content with colorimeter method.
No significant difference in weight gains and feed efficiency among groups were found. Apparent absorption of iron absorption was found to increase with the increasing addition of transgenic rice. Enhancing iron absorption is most profound at single addition of transgenic rice. On the contrary, double calculated transgenic rice addition exhibit most effective to enhance the absorption of zinc. Absorption of calcium were found to increase gradually with increase in the transgenic rice addition.Enhancing absorption of magnesium by transgenic rice addition was most effective among all metals examined. The phosphorus absorption exhibited to increase with increase in transgenic rice incorporation into regular rice and most profound at the single incorporation.

壹、 中文摘要-----------------------------------------------------------------------------------------1
英文摘要-----------------------------------------------------------------------------------------3
貳、 前言-----------------------------------------------------------------------------------------------5
參、 文獻回顧-----------------------------------------------------------------------------------------7
一、米---------------------------------------------------------------------------------------------7
（一）米的背景及種類介紹----------------------------------------------------------7
（二）米的營養及應用----------------------------------------------------------------8
二、花生-----------------------------------------------------------------------------------------12
三、植酸-----------------------------------------------------------------------------------------14
（一）植酸背景介紹------------------------------------------------------------------14
（二）植酸營養效應------------------------------------------------------------------14
（三）植酸及其水解物之功用-----------------------------------------------------18
四、植酸酶-------------------------------------------------------------------------------------20
（一）植酸酶的發展-----------------------------------------------------------------20
（二）植酸酶來源與作用機制----------------------------------------------------20
（三） 影響植酸酶活性之因子---------------------------------------------------23
（四）植酸酶之應用------------------------------------------------------------------25
（五）基因工程改良植酸酶-------------------------------------------------------27
肆、 材料方法與步驟------------------------------------------------------------------------------31
一、 實驗材料---------------------------------------------------------------------------------31
二、 實驗設備---------------------------------------------------------------------------------32
三、 化學藥品---------------------------------------------------------------------------------32
四、 實驗方法---------------------------------------------------------------------------------33
(一) 水稻植酸酶之粗酵素----------------------------------------------------------33
(二) 植酸酵素活性分析--------------------------------------------------------------34
(三) 植酸含量測定--------------------------------------------------------------------36
(四) 發芽米製作-----------------------------------------------------------------------37
(五) 化學成分分析--------------------------------------------------------------------37
(六) 動物實驗--------------------------------------------------------------------------39
(七) 礦物質分析-----------------------------------------------------------------------43
五、統計分析---------------------------------------------------------------------------------46
伍、 結果討論----------------------------------------------------------------------------------------48
一、不同市售稻米植酸含量比較--------------------------------------------------------48
二、糙米在不同發芽時間之植酸酶活性-----------------------------------------------48
三、米漿粉中植酸、蛋白質與油脂組成以及飼料配置----------------------------51
四、動物生長實驗----------------------------------------------------------------------------51
五、餵食米漿粉飼料對礦物質之生物可利用性-------------------------------------53
陸、 結論----------------------------------------------------------------------------------------------60
柒、 參考文獻----------------------------------------------------------------------------------------62
















圖目錄

表一、稻米必需氨基酸(EAA)組成和WHO認定蛋白質氨基酸最佳配比模式( %)-10
表二、穀類蛋白質之生物價、消化率、淨利用率及功效比值----------------------------11
表三、穀類、豆類及其製品與根莖植物、水果中植酸之含量--------------------------16
表四、植酸酶歷史-----------------------------------------------------------------------------------21
表五、不同來源之植酸酶特性-------------------------------------------------------------------22
表六、生產植酸酶之微生物菌種----------------------------------------------------------------29
表七、米漿粉成分表-------------------------------------------------------------------------------42
表八、不同礦物質之石墨原子吸收光譜溫度條件-----------------------------------------45
表九、不同品牌之市售稻米植酸含量---------------------------------------------------------49
表十、在米粉、花生粉和米漿粉中植酸、蛋白質和油脂的含量-----------------------52
表十一、植酸和植酸酶對於進食飼料量、體重和飼料轉換率的影響---------------54
表十二、植酸和植酸酶對鐵和鋅吸收的影響------------------------------------------------55
表十三、植酸和植酸酶對鈣和磷吸收的影響-----------------------------------------------56
表十四、植酸和植酸酶對鎂吸收的影響------------------------------------------------------57



表目錄

圖一、植酸之化學架構----------------------------------------------------------------------------15
圖二、植酸與蛋白質、金屬及澱粉之可能反應--------------------------------------------19
圖三、植酸經植酸酶水解為肌醇及磷酸之水解圖-----------------------------------------24
圖四、植物及黴菌所產之植酸酶降解植酸途徑--------------------------------------------26
圖五A、動物實驗大鼠飼養籠-------------------------------------------------------------------41
圖五B、糞尿分離不銹鋼墊盤--------------------------------------------------------------------41
圖五C、墊盤細部圖--------------------------------------------------------------------------------41
圖六、糙米在不同發芽時間下植酸酶之活性-----------------------------------------------50

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