中国中药杂志

2011, v.36(03) 351-355

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转基因何首乌毛状根生物转化瑞香素的研究
Biotransformation of daphnetin by suspension transgenic hairy roots of Polygonum multiflorum

邓文娟;周良彬;于荣敏;
DENG Wenjuan,ZHOU Liangbin,YU Rongmin(College of Pharmacy,Jinan University,Guangzhou 510632,China)

摘要(Abstract):

目的:利用转基因何首乌毛状根对瑞香素(I)进行生物转化研究,对其生物转化产物进行分离、鉴定,并建立其产物的时效曲线图。方法:将瑞香素投入何首乌毛状根悬浮培养体系中,共培养36 h后,利用TLC和HPLC进行产物检测,硅胶柱层析法分离纯化,核磁共振技术进行结构鉴定,利用HPLC考察共培养时间对转化产物(Ⅱ)、降解产物含量的影响及分析转化产物降解的原因。结果:瑞香素在何首乌毛状根悬浮培养体系中发生了生物转化反应。分离鉴定出一个产物:瑞香素-8-O-β-D-葡萄糖苷(Ⅱ)。何首乌毛状根悬浮培养体系转化生成Ⅱ的最佳共培养时间为36 h,总摩尔转化率为32.11%。蔗糖-水培养基(仅含有蔗糖和水)可以使何首乌毛状根悬浮培养体系转化产率大幅增加,总摩尔转化率达72.44%。初步推断产物(Ⅱ)的降解与MS培养基中金属离子有关。结论:使用何首乌毛状根悬浮培养体系可大量生物合成天然药材中含量极低的活性化合物——瑞香素-8-O-β-D-葡萄糖苷。
Objective: To investigate the biotransformation of daphnetin by suspension transgenic hairy root of Polygonum multiflorum and provide a biotechnological method for large-scale production of the daphnetin-8-O-β-D-glucoside using this new culture system.Method: Daphnetin was added into the media of suspension to culture 36 h.The biotransformation product was detected with TLC and HPLC,and isolated by various chromatographic methods.The influence of co-cultured time on conversion ratio,content of degradation product and the reason for the degradation of product II were investigated using HPLC.Result: One biotransformation product,daphnetin-8-O-β-D-glucoside,was obtained,the optimal co-cultured time in suspension hairy root of P.multiflorum was 36 h with the highest biotransformation molar ratio of 32.11%,the sucrose medium(sucrose-only) can increase the biotransformation molar ratio to 72.44%.The result demonstrated that the degradation products of the product II was induced by the MS medium.Conclusion: The potential application of suspension transgenic hairy root of P.multiflorum in the sucrose-only medium on generating daphnetin-8-O-β-D-glucoside could be prospective.

关键词(KeyWords): 何首乌毛状根;生物转化;瑞香素;瑞香素-8-O-β-D-葡萄糖苷
hairy roots;Polygonum multiflorum;biotransformation;daphnetin;daphnetin-8-O-β-D-glucoside.

Abstract:

Keywords:

基金项目(Foundation): 广东省自然科学基金(04010461)

作者(Authors): 邓文娟;周良彬;于荣敏;
DENG Wenjuan,ZHOU Liangbin,YU Rongmin(College of Pharmacy,Jinan University,Guangzhou 510632,China)

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