摘要
木香烃内酯作为最简单的倍半萜内酯,表现出强大的抗炎和抗肿瘤特性,近年来已被探索作为各种肿瘤的潜在抗癌剂。然而,化学合成和植物提取等获取方法均无法满足可持续的发展需求;因此,利用微生物细胞工厂合成木香烃内酯已成为可行的替代方案。该研究使用大肠杆菌作为宿主生物,以葡萄糖为碳源,过表达吉马烯A合成酶、吉马烯A氧化酶、木香烃内酯合成酶、细胞色素P450还原酶并引入外源甲羟戊酸途径,成功在大肠杆菌BL21(DE3)中构建了木香烃内酯生物合成途径。通过一系列策略,包括筛选表达系统和木香烃内酯合成酶、补充血红素前体、引入代谢工程菌株以及发酵条件优化,最终在摇瓶中产生了6.19 mg/L的木香烃内酯,与起始菌株相比增加了81倍。结果证明了使用微生物细菌细胞工厂合成木香烃内酯的可行性,并为其他倍半萜内酯的合成提供了一种参考策略。
Costunolide,the simplest sesquiterpene lactone,has potent anti-inflammatory and antitumor properties and has been investigated as a potential anticancer agent for various tumors in recent years.However,conventional acquisition methods such as chemical synthesis and plant extraction have proven inadequate to meet the demand for sustainable development.Consequently,the synthesis of costunolide using microbial cell factories has emerged as a viable alternative.This study successfully constructed the costunolide biosynthesis pathway by utilizing Escherichia coli BL21(DE3)as the host organism and glucose as the carbon source,overexpressing the enzyme germacrene A synthase,germacrene A oxidase,costunolide synthase,the cytochrome P450 reductase,and introducing the exogenous mevalonate pathway.Through a series of strategies including screening of expression system and costunolide synthase,supplementation with heme precursors,introduction of metabolically engineered strains,optimization of fermentation conditions,and final produced 6.19 mg/L costunolide in shake flasks,which represents an 81-fold increase compared with the starting strain.The results demonstrated the feasibility of utilizing microbial bacterial cell factories for the synthesis of costunolide and provided a reference strategy for the synthesis of other sesquiterpene lactones.
作者
杨博楠
刘春立
郝云鹏
许广博
刘秀霞
杨艳坤
白仲虎
YANG Bonan;LIU Chunli;HAO Yunpeng;XU Guangbo;LIU Xiuxia;YANG Yankun;BAI Zhonghu(Key Laboratory of Industrial Biotechnology of Ministry of Education,School of Biotechnology,Jiangnan University,Wuxi 214122,China;National Engineering Research Center of Cereal Fermentation and Food Biomanufacturing,Jiangnan University,Wuxi 214122,China;Jiangsu Provincial Engineering Research Center for Bioactive Product Processing,Jiangnan University,Wuxi 214122,China)
出处
《食品与发酵工业》
北大核心
2025年第20期77-83,共7页
Food and Fermentation Industries
基金
国家自然科学青年基金项目(32200092)。
