三分三悬浮培养细胞中苯丙氨酸、鸟氨酸和莨菪碱至东莨菪碱的生物转化

在三分三悬浮培养中分别补充前体苯丙氨酸、鸟氨酸和莨菪碱时,悬浮细胞的生长速率及东莨菪碱的转化率均以补充苯丙氨酸者为最高,分别比未加前体的对照增加50.2％和4.0％。 在上述处理培养两周后于培养液中分别加入70ppm的放线菌酮抑制蛋白质合成时,虽然细胞中东莨菪碱的转化按克干重计算均有明显增加,但是由于细胞生长受强烈抑制不利于东莨菪碱的总的转化,按每瓶总干重计算的转化率均比对照低。 在分别补充三种前体并培养两周后使营养缺乏时,细胞生长比对照稍有降低,但细胞中东莨菪碱的转化却显著增加。特别是补充莨菪碱并使营养缺乏时,大大利于莨菪碱转化为东莨菪碱,转化率高达27.4％。既降低了培养成本又提高了东莨菪碱产量,有利于未来的工业生产。

BIOTRANSFORMATION OF PHENYLALANINE, ORNITHINE AND HYOSCYAMINE TO SCOPOLAMINE IN THE SUSPENSION CULTURE OF ANISODUS ACUTANGULUS CELLS

When the suspension cultures of Anisodus acutangulus were supplemented with precursors of either phenylalanine. ornithine or hyocyamine. phenylalanine was found to be most effective in promoting both the cell growth and its conversion to scopolamine. The growth rate of the cultures adding phenylalanine as precursor was 50. 2% higher than that without phenylalanins and the ratio of conversion was 4.0%Upon addition of actidionc at the concentration of 30 ppm to the two-week-old cultures, protein synthesis was inhibited. Under such conditions, although the ratio of conversion from various precursors to scopolamine in the cells was increased markedly in terms of gram dry weight, the total scopolamine production per culture flask actually was decreased due to the fact that cell growth. was strongly inhibited by actidione, resulting on lower dry weight yield of sco-polamine than the control.Cell cultures supplimented with three precursors that were allowed to grow for two weeks and then nutritionally depleted had lower cell growth rate than the control, but their conversion rate from precursors to scopolamine was actually increased significantly, especially in the case where addition of hyoscya-mine and depletion of nutrition were done at the same time. Under such conditions, the conversion rate from hyoscyamine to scopolamine was as high as 27.4%. These findings showed that the rate of conversion from precursors to scopolamine could be raised while reducing the production (culturing) cost may be applied beneficially to scopolamine production on industrial scale.