【摘要】 目的应用悬浮培养转基因西洋参冠瘿组织生物合成熊果苷。方法向预培养20 d的西洋参冠瘿组织培养物中加入60 m g/mL对苯二酚溶液,共培养60 h后终止转化;通过TLC和HPLC方法检测产物,使用柱色谱法分离纯化产物,利用光谱数据对其结构进行鉴定。HPLC定量分析熊果苷产生的动力学变化曲线。结果产物同时在冠瘿组织培养物和培养基存在,并经过提取、分离得到其纯品,经理化性质和谱学方法将其鉴定为4-羟基苯-β-D-吡喃葡萄糖苷(熊果苷)。摩尔转化率在共培养60 h后达到最高(88.4%),同时,培养物和培养基中产物的量(63.69 m g/瓶,1.86 m g/瓶)以及产物的分泌百分率都达到最高(2.92%)。结论在国内外首次将转基因冠瘿组织用于生物转化系统并成功地得到具有多种药理活性的熊果苷。更多还原

【Abstract】 Objective To investigate the biosynthesis of arbutin by suspension culture of crown gall of transgenic Panax quinquefolium. Methods Hydroquinone in methanol(60 mg/mL) was added to the medium of the crown gall of P.quinquefolium after precultured for 20 d,then they were co-cultured for another 60 h.The product was isolated and purified by column chromatography and its structure was identified by physicochemical properties and spectroscopic data.The dynamic curve of biotransformation was investigated by quantative analysis of arbutin with HPLC.Results The product could be obtained from both of the culture and medium,which was isolated and elucidated as 4-hydroxyphenyl-β-D-glucopyranoside(arbutin).After co-cultured for 60 h,the mole conversion ratio of hydroquinone is 88.4%,the product contents in culture and medium are 63.69 mg/flask and 1.86 mg/flask,respectively,and the excretion ratio of arbutin reaches the highest(2.92%).Conclusion It’s the first time around the world that the crown gall of transgenic P.quinquefolium is used as a biotransformation system and arbutin which shows varied pharmacological activites have been got successfully.更多还原