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題名 | 毛細管電泳在蜂王漿蛋白質分析上之應用=The Application of Capillary Electrophoresis on the Characterization of Protein in Royal Jelly |
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作者 | 李安玲; 葉美吟; 溫惠美; 陳景川; 林貞信; 黃文瑛; Lee, Ann-lyne; Yeh, Mei-ing; Wen, Hwei-mei; Chern, Jiing-chuan; Lin, Jenshinn; Hwang, Wen-ing; |
期刊 | 藥物食品分析 |
出版日期 | 19990300 |
卷期 | 7:1 1999.03[民88.03] |
頁次 | 頁73-82 |
分類號 | 438.77 |
語文 | chi |
關鍵詞 | 蛋白質; 蜂王漿; 毛細管電泳; Protein; Royal jelly; Capillary; Electrophoresis; |
中文摘要 | 本研究採本省中南部以生產蜂王漿為主之義大利蜂種所生產之新鮮蜂王漿,進行 蛋白質特性之探討。蜂王漿中總氮含量為 2.46 %,其中來自總胺基酸的氮含量為 2.34 % ;而來自游離胺基酸的氮含量為 0.11 %,胺基態氮的含量為 0.20 %,顯示蜂王漿中的含 氮化合物大部分是以蛋白質的型態存在。蜂王漿經離心、沉澱和透析等分離及純化後,可得 65 ± 5 %之水溶性蛋白質,再經 DEAE-Sephacel 離子交換管柱分離, 得到二部分之區分 物質,F1 和 F2。 將 F1 和 F2 分別以 SDS-PAGE 電泳分析,F1 部份含二個染色帶,分子 量分別為 50 及 44KDa,F2 部分則僅含一個染色帶,分子量為 55KDa。 毛細管電泳之分析 模式,由於分離機制與解析度各不相同,可將蜂王漿蛋白質分離成不同區分。 將 F1 和 F2 經毛細管區帶電泳分析,F1 出現四個吸收蜂,F2 則有二個吸收峰;而經毛細管凝膠電泳分 析,F1 部分出現二個吸收峰,分子量分別為 59 及 73KDa,F2 則僅出現一個吸收峰,分子 量為 118KDa; 但經毛細管等電聚焦來分離 F1 和 F2,F1 則出現六個吸收峰,等電點分別 為 6.9,6.7,6.3,5.9,5.7 及 5.5,F2 出現二個吸收峰,等電點為 4.8 及 4.7。 |
英文摘要 | The objective of this study was to characterize the protein fractions in royal jelly made from Apis mellifera ligustica from middle and southern areas of Taiwan. The total nitrogen content of fresh royal jelly was 2.46 %, and the total amino acid nitrogen was 2.34%, suggesting that the nitrogen compound in royal jelly was mostly derived from protein. The nitrogen content of free amino acid in royal jelly was 0.11%, and the amino type nitrogen was 0.20 %, indicating that the protein in royal jelly existed mainly in the form of large moleculars. To characterize the protein, royal jelly was dissolved in 0.1 M phosphate buffer (pH7.0) , followed by centrifugation, ammonia sulfate precipitation and dialysis to separate the protein into water soluble and water insoluble fractions. Water soluble fraction accounts for more than 60 % of the total protein in royal jelly, and was further investigated by DEAE-Sephacel, SDSPAGE and capillary electrophoresis. By DEAE-Sephacel, two fraction peaks (F1 and F2) were identified and collected. By SDS-PAGE, F1 fraction was further separated into two bands, and the molecular weight was determined to be 50 KDa and 44 KDa, whereas F2 fraction was shown to have only one band with molecular weight of 55 KDa. By capillary zone electrophoresis, four poorly-separated peaks were observed in F1 fraction, and two well-separated peaks in F2 fraction. By capillary gel electrophoresis, two peaks were identified in F1 fraction, of which the molecular weight was estimated to be 59 KDa and 73 KDa. By contrast, only one peak was identified in F2 fraction, of which the molecular weight was estimated to be 118 KDa. However, by capillary isoelectric focusing, 6 peaks were identified in F1 fraction with pI of 6.9,6.7,6.3,5.9,5.7 and 5.5, respectively. Of that, pI 4.8 and 4.7 were identified in F2 fraction. |
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