摘要
天麻苷元是我国名贵中药材天麻中的一种重要活性成分,也是天麻素生物合成的主要前体物质,具有抗炎、抗肿瘤以及保护脑缺血等多种药理活性,其作为主要成分的医药产品越来越受人们的青睐。当前天麻苷元的获取主要以天然提取和化学合成为主,但它们都存在一些限制其长远应用的不足,如野生和栽培天麻资源有限,化学合成需步骤多、时间长、条件苛刻的反应,且反应副产物多、反应三废不好处理等,因此如何人工制备产量大、纯度高的此类物质,已成为医药研究工作者一个亟待解决的问题。该研究以微生物代谢工程理论为指导,采用基因工程相关技术,将ThiH,pchF和pchC 3个基因导入大肠杆菌,使其成为利用L-酪氨酸合成天麻苷元的工程菌,并利用摇瓶发酵手段对该工程菌发酵生成天麻苷元的条件进行探讨。实验结果显示,该工程菌在IPTG浓度为0.5 mmol·L^(-1)、诱导温度为15℃、转化温度为40℃时,可将含200 mg·L^(-1)L-酪氨酸的M9Y培养基转化生成(69±5)mg·L^(-1)天麻苷元,这一结果为日后进一步扩大发酵规模,从而既经济又高效地生产天麻苷元奠定了基础。
As an important active ingredient in the rare Chinese herb Gastrodiae Rhizoma and also the main precursor for gastrodin biosynthesis,4-hydroxybenzyl alcohol has multiple pharmacological activities such as anti-inflammation,anti-tumor,and anti-cerebral ischemia.The pharmaceutical products with 4-hydroxybenzyl alcohol as the main component have been increasingly favored.At present,4-hydroxybenzyl alcohol is mainly obtained by natural extraction and chemical synthesis,both of which,however,exhibit some shortcomings that limit the long-term application of 4-hydroxybenzyl alcohol.The wild and cultivated Gastrodia elata resources are limited.The chemical synthesis requires many steps,long time,and harsh reaction conditions.Besides,the resulting by-products are massive and three reaction wastes are difficult to treat.Therefore,how to artificially prepare 4-hydroxybenzyl alcohol with high yield and purity has become an urgent problem facing the medical researchers.Guided by the theory of microbial metabolic engineering,this study employed the genetic engineering technologies to introduce three genes ThiH,pchF and pchC into Escherichia coli for synthesizing 4-hydroxybenzyl alcohol with L-tyrosine.And the fermentation conditions of engineering strain for producing 4-hydroxybenzyl alcohol in shake flask were also discussed.The experimental results showed that under the conditions of 0.5 mmol·L^(-1)IPTG,15℃induction temperature,and 40℃transformation temperature,M9 Y medium containing 200 mg·L^(-1)L-tyrosine could be transformed into(69±5)mg·L^(-1)4-hydroxybenzyl alcohol,which has laid a foundation for producing 4-hydroxybenzyl alcohol economically and efficiently by further expanding the fermentation scale in the future.
作者
徐德宏
保晓清
吴细文
邢宇
谭朝阳
XU De-hong;BAO Xiao-qing;WU Xi-wen;XING Yu;TAN Chao-yang(Biological Engineering Laboratory,College of Pharmacy,Hunan University of Chinese Medicine,Changsha 410208,China)
出处
《中国中药杂志》
CAS
CSCD
北大核心
2022年第4期906-912,共7页
China Journal of Chinese Materia Medica
基金
国家自然科学基金项目(82104324)
湖南省自然科学基金项目(2019JJ50444)
湖南中医药大学生物工程重点学科项目(校行科字[2018]3号)
湖南中医药大学校级科研基金项目(2020XJJJ030)
湖南省教育厅资助科研项目(19C1407)。
关键词
代谢工程
天麻苷元
L-酪氨酸
大肠杆菌
metabolic engineering
4-hydroxybenzyl alcohol
L-tyrosine
Escherichia coli
