中国中药杂志2025，Vol.50Issue(5)：1187-1194,8.DOI:10.19540/j.cnki.cjcmm.20241215.102
大肠杆菌全细胞转化1-(4-羟基苯酚)-乙醇合成中药活性物质青心酮的研究
Synthesis of active substance 3,4-dihydroxyacetophenone from traditional Chinese medicine using Escherichia coli whole-cell bioconversion of 1-(4-hydroxyphenol)-ethanol
摘要
Abstract
The main active compound,3,4-dihydroxyacetophenone(3,4-DHAP),in the leaves of Ilex pubescens var.glaber,exhibits various pharmacological activities,including vasodilation and heart protection.Currently,natural extraction and chemical synthesis are the primary methods for obtaining 3,4-DHAP,but both approaches have inherent challenges.To address these problems,this study explored the whole-cell bioconversion of 1-(4-hydroxyphenol)-ethanol to 3,4-DHAP using recombinant Escherichia coli,cultivated in a green,cost-effective medium at room temperature and atmospheric pressure.Firstly,this study successfully constructed recombinant E.coli S1,which contained only the HpaBC gene,and recombinant E.coli S3,which contained both the Hped and HpaBC genes.The ability of S1 and S3 to synthesize 3,4-DHAP from their respective substrates was then evaluated through whole-cell bioconversion.Based on these results,the effects of four factors,i.e.,substrate concentration,IPTG concentration,induction temperature,and transformation temperature,on the whole-cell bioconversion yield of S3 were investigated using an orthogonal experiment.The results showed that the factors influenced the yield in the following order:transformation temperature>induction temperature>IPTG concentration>substrate concentration.The optimal conditions were found to be a transformation temperature of 35℃,IPTG concentration of 0.1 mmol·L-1,induction temperature of 25℃,and substrate concentration of 10 mmol·L-1.Finally,the effect of transformation time on the yield of 3,4-DHAP was further examined under the optimal conditions.The results indicated that as the transformation time increased,the yield of 3,4-DHAP steadily increased.The highest yield of 260 mg·L-1 with a productivity of 17%was achieved after 72 hours of transformation.In conclusion,this study successfully achieved the whole-cell bioconversion of 1-(4-hydroxyphenol)-ethanol to 3,4-DHAP using recombinant E.coli for the first time,laying the groundwork for further optimization and development of the biosynthesis of 3,4-DHAP.关键词
青心酮/重组大肠杆菌/全细胞转化/1-(4-羟基苯酚)-乙醇/1-(4-羟基苯酚)-乙醇脱氢酶/4-羟基苯乙酸3-单加氧酶Key words
3,4-dihydroxyacetophenone/recombinant Escherichia coli/whole-cell bioconversion/1-(4-hydroxyphenol)-ethanol/1-(4-hydroxyphenol)-ethanol dehydrogenase/4-hydroxy phenylacetate 3-hydroxylase引用本文复制引用
袁希伟,田艳秋,王文宇,张亚伦,徐德宏..大肠杆菌全细胞转化1-(4-羟基苯酚)-乙醇合成中药活性物质青心酮的研究[J].中国中药杂志,2025,50(5):1187-1194,8.基金项目
国家自然科学基金项目(82104324) （82104324）
湖南省教育厅优秀青年项目(23B0358) （23B0358）
湖南中医药大学"十四五"重点学科(生物工程学科)项目(校行发规字[2023]2号) （生物工程学科）
湖南中医药大学研究生创新课题(2023CX138) （2023CX138）
