【摘要】 三分三浸没培养细胞的生长速率是1.5克千重/升/天,为三分三细胞悬浮培养的3倍,为三分三固体静态培养的10倍多。浸没培养细胞中莨菪碱(0.203毫克/克干重)和东莨菪碱(0.178毫克/克干重)含量仅稍高于悬浮培养和固体培养物。但在培养12天之后的浸没培养中补充以苯丙氨酸(5毫克分子/升)和激动素(0.1毫克/升)时,不仅培养细胞的生长速率有所增加而且培养细胞中莨菪碱含量可高达0.217毫克/克干重和东莨菪碱含量高达0.412毫克/克干重。两种生物碱的含量分别为未补充这些化合物的对照的1.1和2.3倍。三分三浸没培养的较适收获期是在浸没培养的第14天。从培养细胞中分离到莨菪碱和东莨菪碱单体。经纯化和重结晶之后被薄层层析、熔点和紫外、红外、核磁共振等光谱鉴定,证明了它们确系茛菪碱和东莨菪碱结晶。研究结果表明可用三分三细胞发酵培养法生产莨菪碱和东茛菪碱。更多还原

【Abstract】 The growth rate of Anisodus acutangulus cells in the submergence culture was 1.5 g dry wt/1/day, a rate 3 times as that in the suspension culture and more than 10 times over that of the solid static culture. However, the contents of hyoscyamine (0.203 mg/g dry wt) and scopolamine (0.178 mg/g dry wt) in submergence culture were only slightly higher than those in the two other cultures. But when the 12-day-old submergence culture was supplemented with phenylalanine (5 m mol/l) and kinetin (0.1mg/l), it was observed that not only the cell growth rate was increased but also the cellular content of hyoscyamine was raised to a level of 0.217 mg/g dry wt, and that scopolamine to 0.412 mg/g dry wt. The contents of these two alkaloids represented 1.1 and 2.3 times respectively the value of the culture without phenylalanine and kinetin supplements. The optimum date for harvesting the A. acutangulus culture was on the 14th day of the culture. The monomers of hyoscyamine and scopolamine isolated from the cultured cells were purified and recrystalized, and then identifited as the two compounds in question by the thin-layer chromatography, melting point determination, and ultra-violet, infra-red and nuclear magnetic resonance spectra. In this paper, we also summarize and discuss the results of A. acutangulus culture experiments performed in the past 8 years. Our finding seems to indicate that following a pilot production trial, the tissue culture method could well be employed to produce hyoscyamine and scopolamine from A. acutangulus cells on an industrial scale.更多还原