Engineering cellular dephosphorylation boosts (+)-borneol production in yeast.
10.1016/j.apsb.2024.12.039
- Author:
Haiyan ZHANG
1
;
Peng CAI
2
;
Juan GUO
1
;
Jiaoqi GAO
2
;
Linfeng XIE
2
;
Ping SU
1
;
Xiaoxin ZHAI
2
;
Baolong JIN
1
;
Guanghong CUI
1
;
Yongjin J ZHOU
2
;
Luqi HUANG
1
Author Information
1. State Key Laboratory for Quality Ensurance and Sustainable Use of Dao-di Herbs, National Resource Centre for Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing 100700, China.
2. Division of Biotechnology, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian 116023, China.
- Publication Type:Journal Article
- Keywords:
Borneol;
DPP1;
LPP1;
Monoterpene;
Nudix hydrolase;
Phosphatase
- From:
Acta Pharmaceutica Sinica B
2025;15(2):1171-1182
- CountryChina
- Language:English
-
Abstract:
(+)-Borneol, the main component of "Natural Borneol" in the Chinese Pharmacopoeia, is a high-end spice and precious medicine. Plant extraction cannot meet the increasing demand for (+)-borneol, while microbial biosynthesis offers a sustainable supply route. However, its production was extremely low compared with other monoterpenes, even with extensively optimizing the mevalonate pathway. We found that the key challenge is the complex and unusual dephosphorylation reaction of bornyl diphosphate (BPP), which suffers the side-reaction and the competition from the cellular dephosphorylation process, especially lipid metabolism, thus limiting (+)-borneol synthesis. Here, we systematically optimized the dephosphorylation process by identifying, characterizing phosphatases, and balancing cellular dephosphorylation metabolism. For the first time, we identified two endogenous phosphatases and seven heterologous phosphatases, which significantly increased (+)-borneol production by up to 152%. By engineering BPP dephosphorylation and optimizing the MVA pathway, the production of (+)-borneol was increased by 33.8-fold, which enabled the production of 753 mg/L under fed-batch fermentation in shake flasks, so far the highest reported in the literature. This study showed that rewiring dephosphorylation metabolism was essential for high-level production of (+)-borneol in Saccharomyces cerevisiae, and balancing cellular dephosphorylation is also helpful for efficient biosynthesis of other terpenoids since all whose biosynthesis involves the dephosphorylation procedure.
