西洋参、人参的细胞大量培养及其皂甙成分的分离和鉴定
周立刚
学位类型硕士
导师郑光植
1989
学位授予单位中国科学院昆明植物研究所
学位授予地点中国科学院昆明植物研究所
学位专业植物学
关键词西洋参 人参 悬浮培养 发酵培养 人参皂甙 寡糖
摘要西洋参和人参细胞悬浮培养结果表明,其合成人参皂甙的高峰在细胞生长的对数期稍后出现;对人参、西洋参细胞发酵培养、液体静止培养和悬浮培养的比较研究表明,生长速率和生物量均以悬浮培养最高;在发酵罐培养中,PH值稳定的培养比PH变化的培养,其皂甙含量、生长速率和生物量均要高;在相同的通气速率(VVm)情况下,无细胞培养液的液膜容量传质系数(kLa)值最大,人参细胞培养液次之,西洋参细胞培养液的kLa值最小。对西洋参还进行了其它一系列大量培养的研究:当培养液中硝态氮含量提高一倍而去掉氨态氮时,细胞生长速率和皂甙含量分别比对照提高65.1%和166.2%;当提高培养基的渗透压时,西洋参细胞培养物的皂甙含量为1.58%,为对照的3倍;西洋参细胞发酵培养的适宜搅拌速度为每分钟60转。黑节草寡糖和人参寡糖均有利于西洋参悬浮培养细胞的生长和皂甙含量的提高,尤其能增加Rg组皂甙的含量,含量分别对照的1.78倍和2.83倍。以上两类寡糖还能影响Rg组皂甙与Rb组皂甙之比例。从西洋参细胞培养物中,分离了其有效成分人参皂甙单体R-A,R-B和R-C,并对其进行了薄层层析(TLC),核磁其振碳谱(~(13)CNMR),电子轰击质谱(εl-MS),红外(IR),以及旋光、熔点和元素分析的测定,证明它们分别为人参皂甙Rbl,Re和Rgl。从TLC可知细胞培养物的皂甙组分几乎和原植物一致。本研究为将来进一步工业化生产提供了依据。
其他摘要Ginsenosides formation slightly behind the growth curve in suspension culture of quinquefolium and ginseng cells. The growth rate and biomass of suspension culture were the greatest in static liquid culture, suspension culture and fermentation culture. The fermentation culture of quinquefolium cells with a fixed PH value was better than the culture with a changing PH value by itself. The volumetric oxygen transfer coefficient (Kla) values measured in the cultured broths of ginseng and quinquefolium cells were far smaller than that in the fresh medium. These data suggest that the high viscosity of the cultured broths can be one of the problems in scaling up this system. It was found that MS medium minus NH_4NO_3 and with added KNO_3 gave a higher growth ratio and saponin content than regular MS medium or MS minus KNO_3. The growth ratio and saponin content with the medium only added KNO_3 were increased 65.1% and 166.2% separately when compared with the regular MS medium. The saponin content reached 1.58%which was 3 times as high as the control when we applied an osmotic stress by the addition of mannitol to the nutrient medium in fermentation culture of quinquefolium cells. An appropriate agitating cpeed was 60rpm in fermentation culture of quinquefolium cells. Oligosaccharins of Panax ginseng and Dendrum canditum increased both saponin content and growth ratio. Especially, the content of Rg group saponins was well stimulated, which was 1.78 times (added ginseng oligosaccharin) and 2.83 times (added canditum oligosaccharin) as high as the control. Ginsenoside Rb1, Re and Rg1 from quinquefolium cell cultures were isolated and identificated by TLC, ~(13)CNMR, mp, IR, [a]_D~OC, EI-MS and elemental analysis. Saponin compositions of the cultures of quinquefolium cells were almost same with those of the cultivated plants. Studies cn the mass culture of quinquefolium and ginseng cells could well be employed to produce ginsenosides on an industrial scale in the fulture.
页数74
语种中文
文献类型学位论文
条目标识符http://ir.kib.ac.cn/handle/151853/852
专题昆明植物所硕博研究生毕业学位论文
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周立刚. 西洋参、人参的细胞大量培养及其皂甙成分的分离和鉴定[D]. 中国科学院昆明植物研究所. 中国科学院昆明植物研究所,1989.
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