Construction of Saccharomyces cerevisiae cell factory for efficient biosynthesis of ferruginol.
10.19540/j.cnki.cjcmm.20241115.104
- Author:
Mei-Ling JIANG
1
;
Zhen-Jiang TIAN
2
;
Hao TANG
1
;
Xin-Qi SONG
3
;
Jian WANG
4
;
Ying MA
4
;
Ping SU
4
;
Guo-Wei JIA
5
;
Ya-Ting HU
6
;
Lu-Qi HUANG
7
Author Information
1. School of Pharmacy, Nanjing University of Chinese Medicine Nanjing 210023, China National Resource Center for Chinese Materia Medica, China Academy of Chinese Medical Sciences Beijing 100700, China.
2. School of Traditional Chinese Medicine, Capital Medical University Beijing 100069, China.
3. National Resource Center for Chinese Materia Medica, China Academy of Chinese Medical Sciences Beijing 100700, China School of Pharmacy, Jiangxi University of Chinese Medicine Nanchang 330004, China.
4. National Resource Center for Chinese Materia Medica, China Academy of Chinese Medical Sciences Beijing 100700, China.
5. National Resource Center for Chinese Materia Medica, China Academy of Chinese Medical Sciences Beijing 100700, China School of Pharmacy, Jiangsu University Zhenjiang 212013, China.
6. School of Traditional Chinese Medicine, Capital Medical University Beijing 100069, China State Key Laboratory for Quality Ensurance and Sustainable Use of Dao-di Herbs Beijing 100700, China.
7. National Resource Center for Chinese Materia Medica, China Academy of Chinese Medical Sciences Beijing 100700, China State Key Laboratory for Quality Ensurance and Sustainable Use of Dao-di Herbs Beijing 100700, China.
- Publication Type:Journal Article
- Keywords:
Saccharomyces cerevisiae;
diterpenoids;
ferruginol;
metabolic engineering
- MeSH:
Saccharomyces cerevisiae/genetics*;
Diterpenes/metabolism*;
Metabolic Engineering;
Fermentation;
Abietanes
- From:
China Journal of Chinese Materia Medica
2025;50(4):1031-1042
- CountryChina
- Language:Chinese
-
Abstract:
Diterpenoid ferruginol is a key intermediate in biosynthesis of active ingredients such as tanshinone and carnosic acid.However, the traditional process of obtaining ferruginol from plants is often cumbersome and inefficient. In recent years, the increasingly developing gene editing technology has been gradually applied to the heterologous production of natural products, but the production of ferruginol in microbe is still very low, which has become an obstacle to the efficient biosynthesis of downstream chemicals, such as tanshinone. In this study, miltiradiene was produced by integrating the shortened diterpene synthase fusion protein,and the key genes in the MVA pathway were overexpressed to improve the yield of miltiradiene. Under the shake flask fermentation condition, the yield of miltiradiene reached about(113. 12±17. 4)mg·L~(-1). Subsequently, this study integrated the ferruginol synthase Sm CYP76AH1 and Sm CPR1 to reconstruct the ferruginol pathway and thereby realized the heterologous synthesis of ferruginol in Saccharomyces cerevisiae. The study selected the best ferruginol synthase(Il CYP76AH46) from different plants and optimized the expression of pathway genes through redox partner engineering to increase the yield of ferruginol. By increasing the copy number of diterpene synthase, CYP450, and CPR, the yield of ferruginol reached(370. 39± 21. 65) mg·L~(-1) in the shake flask, which was increased by 21. 57-fold compared with that when the initial ferruginol strain JMLT05 was used. Finally, 1 083. 51 mg·L~(-1) ferruginol was obtained by fed-batch fermentation, which is the highest yield of ferruginol from biosynthesis so far. This study provides not only research ideas for other metabolic engineering but also a platform for the construction of cell factories for downstream products.
