论文标题:大豆异黄酮组合物的制备及其生物合成的初步研究
论文作者
论文导师 游松,论文学位 硕士,论文专业 微生物与生化药学
论文单位 沈阳药科大学,点击次数 100,论文页数 66页File Size3016K
2004-06-01论文网 http://www.lw23.com/lunwen_316474542/
Isoflavone;isolation;compisotion;HPLC;isoflavone synthase
本文利用有机溶剂萃取，大孔吸附树脂柱、氧化铝柱、硅胶柱等柱层析，并通过两次HPLC制备，得到了三种化合物纯品，经光谱解析，结构确认为大豆苷、黄豆苷和染料木苷。考查了精制大豆异黄酮组合物的制备工艺，并纯化得到了高纯度大豆异黄酮组合物，液相检测精制大豆异黄酮组合物中大豆苷、黄豆苷与染料木苷之和在95％以上。 建立了大豆异黄酮总苷含量测定方法，进行了方法学考查，结果证明该方法适用于大豆异黄酮总苷的含量测定。对精制大豆异黄酮组合物、不同豆类和大豆不同生长时期各组织的总异黄酮苷进行了含量测定。 根据文献报道的大豆异黄酮合成酶的蛋白和基因序列，用软件设计特异性引物，用RT—PCR方法从大豆幼苗中克隆了大豆异黄酮合成酶cDNA，该片断与文献报道完全一致，大小为1127bp。
Isoflavone composition was prepared by extraction in organic solvent and purification with column chromatography.(macroporous resin, aluminum oxide and silica gel). Three purified compounds obtained from the soybean after two procedure of HPLC preparation were identified by spectral analysis, and they were daidzin、 glycitin and genistin. The preparation method for Isoflavone composition was studied. It was found that the contents of daidzin、 glycitin and genistin was added to more than 95% of all the purifications through the determination of HPLC.The method for the determination of isoflavone was established and evaluated. The results came to the conclusion that the method was suitable for the determination of isoflavone. The isoflavone in purified isoflavone composition, different kinds of beans and soybean tissues in different period of growth was determined.Based on the data of known isoflavone synthase 1, the specific primers were designed. The total RNA was extracted from the soybean seedlings. The core sequence (1127bp) of isoflavone synthase 1 cDNA was cloned with RT-PCR method, which was totally the same with the part of our expected target.
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