中国中药杂志2026，Vol.51Issue(7)：1882-1890,9.DOI:10.19540/j.cnki.cjcmm.20260106.401
高效合成小白菊内酯衍生物kauniolide酿酒酵母细胞工厂构建
Construction of efficient yeast cell factory for kauniolide,a parthenolide derivative
摘要
Abstract
Kauniolide,a parthenolide derivative,serves as a key biosynthetic intermediate for various guaianolide-type sesquiterpenoids,such as agrabin,lactucin,and lactupicrin.However,its chemical synthesis is often hampered by demanding reaction conditions and high reagent consumption,which severely limit further development and application.To address this,this study employed systematic metabolic engineering strategies to construct an efficient yeast cell factory for kauniolide production.First,the biosynthetic pathway genes for costunolide—the direct precursor of kauniolide(HaGAS,TpGAO,TpCOS,AaCPR)—were heterologously expressed in a lab-engineered yeast chassis strain with high farnesyl pyrophosphate(FPP)production.This initial strain produced costunolide at a titer of 6.1 mg·L-1.Subsequently,the co-expression of AaADH1,AaALDH1,and AaCYB5 was implemented to enhance the germacrene A acid synthesis and promote electron transfer,thereby boosting the catalytic efficiency of the key cytochrome P450 enzymes.This modification increased the costunolide titer to 71.5 mg·L-1.To optimize the conversion of costunolide to kauniolide,a promoter compatibility screen for the TpKLS gene was conducted.Among the tested promoters(PGAL1,PTDH3,PTEF1,PTPI1,PsptGAL2,and PskGAL2),the strain harboring the PskGAL2-driven TpKLS construct showed the highest performance,achieving a kauniolide titer of 28.2 mg·L-1.Furthermore,to further enhance the pathway flux,the catalytic efficiency of the TpKLS enzyme was improved through semi-rational design coupled with computer-aided design.The best-performing mutant,TpKLSV204R,exhibited a 2.7-fold higher catalytic activity compared to the wild-type enzyme.The final engineered strain,when cultivated in shake-flask fermentation,produced costunolide and kauniolide at titers of 35.0 and 71.1 mg·L-1,respectively.The kauniolide titer represents the highest level reported to date in a yeast system.In conclusion,this study successfully constructed an efficient yeast cell factory for kauniolide by reconstructing and optimizing its heterologous biosynthetic pathway.It provides a solid technical foundation and a valuable reference for the scalable biosynthesis of kauniolide and the exploration of its downstream derivatives.关键词
kauniolide/倍半萜内酯/TpKLS/功能改造/代谢工程Key words
kauniolide/sesquiterpene lactone/TpKLS/functional modification/metabolic engineering引用本文复制引用
韩洋,谭洪虎,苏平,黄璐琦..高效合成小白菊内酯衍生物kauniolide酿酒酵母细胞工厂构建[J].中国中药杂志,2026,51(7):1882-1890,9.基金项目
中央本级重大增减支项目(2060302) （2060302）
中国中医科学院科技创新工程项目(CI2023E002,CI2024C004YN) （CI2023E002,CI2024C004YN）
