文献类型：期刊文献

英文题名：Sustainable high-titer production of β-elemene in Yarrowia lipolytica via modular engineering, pathway refactoring, and a chimeric germacrene A synthase

作者：Liu, Shun-Cheng[1];Xu, Longxing[2];Sun, Yuejia[1];Zhang, Lei[2];Zhang, Weichen[1];Xu, Jing[2];Liu, Pengcai[1,2];Shi, Fan[1];Tian, Faming[1];Xu, Hong[1];Song, Xiaoming[2];Yuan, Lijie[2];Hua, Qiang[3];Qi, Yi-Ke[4];Liu, Feng[3];Sun, Liangdan[1,5,6]

机构：[1]North China Univ Sci & Technol, Hlth Sci Ctr, Hebei Key Lab Med Engn & Integrated Utilizat Salin, Hebei Adm TCM Key Lab Qual Control Salt Alkali Res, 21 Bohai Ave, Tangshan 063210, Peoples R China;[2]North China Univ Sci & Technol, Sch Basic Med Sci, Tangshan Key Lab Preclin & Basic Res Chron Dis, Hebei Key Lab Chron Dis, 21 Bohai Ave, Tangshan 063210, Peoples R China;[3]East China Univ Sci & Technol, State Key Lab Bioreactor Engn, 130 Meilong Rd, Shanghai 200237, Peoples R China;[4]Hebei Univ Sci & Technol, Coll Food Sci & Biol, 26 Yuxiang St, Shijiazhuang 050018, Peoples R China;[5]North China Univ Sci & Technol, Affiliated Hosp, Tangshan 063000, Peoples R China;[6]Anhui Med Univ, Dept Dermatol, Affiliated Hosp 1, Hefei 230032, Peoples R China

年份：2026

卷号：528

外文期刊名：CHEMICAL ENGINEERING JOURNAL

收录：;EI(收录号：20260219872620);WOS:【SCI-EXPANDED(收录号:WOS:001662232600001)】；

基金：This study was supported by the National Natural Science Foundation of China (No. 22508086, 22501080) ; Hebei Provincial Natural Science Foundation Projects (B2022209030, B2025208076) ; Hebei Provincial Key R & D Program Project (23372505D) ; Scientific Research Project of Colleges and Universities in Hebei Province (BJ2025069) ; Medical-Engineering Integration Project of North China University of Science and Technology (ZD-YG-202423) ; Hebei Provincial Traditional Chinese Medicine Research Project (2025434) ; 2024 Innovation and Entrepreneurship Project of North China University of Science and Technology (329, T2023061) , Hebei Province Major Science and Technology Support Program (252W2502D) , and partially supported by the Key Project for Synthetic Biology of Shanghai (24HC2810800) .

语种：英文

外文关键词：beta-Elemene; Yarrowia lipolytica; Protein engineering; Promoter screening; Pathway refactoring

摘要：(3-Elemene, a pharmacologically active sesquiterpene, holds promise in anti-tumor therapy, yet its plant extraction remains limited by low yield, purification challenges, and environmental concerns. In this study, we engineered the oleaginous yeast Yarrowia lipolytica for high-level (3-elemene production by integrating promoter engineering, protein design, pathway refactoring, and subcellular compartmentalization. First, we identified an endogenous promoter, PTMAL, superior to the well-known strong promoter PTEF1 in driving germacrene A synthase (GAS) expression. A hybrid promoter PUAS1B8TMAL(250) further enhanced production, yielding 156.65 +/- 2.47 mg/L (3-elemene in strain EME-14. To enhance GAS activity, structure-based engineering was conducted on a newly identified GAS from Helianthus annuus L. Through this approach, we pinpointed the H3 domain and residue V436 as critical for its enzymatic activity. Consequently, an optimized chimeric GAS (L1H2H3-V436LH4) increased (3-elemene titer to 563.39 +/- 5.09 mg/L, outperforming the previously reported GAS variants. Subsequent peroxisomal localization of the mevalonate pathway combined with optimization of lipid metabolic pathway boosted production to 3584.18 +/- 87.30 mg/L in strain EME-82, representing a 106.86-fold increase over the initial strain EME-01. Finally, scale-up in a 5-L bioreactor achieved the highest titer of 52.53 g/L (3-elemene. This integrated approach offers an efficient and sustainable strategy for natural product synthesis in Y. lipolytica.