Engineering cellular dephosphorylation boosts (+)-borneol production in yeast

IF 14.7 1区 医学 Q1 PHARMACOLOGY & PHARMACY
Haiyan Zhang , Peng Cai , Juan Guo , Jiaoqi Gao , Linfeng Xie , Ping Su , Xiaoxin Zhai , Baolong Jin , Guanghong Cui , Yongjin J. Zhou , Luqi Huang
{"title":"Engineering cellular dephosphorylation boosts (+)-borneol production in yeast","authors":"Haiyan Zhang ,&nbsp;Peng Cai ,&nbsp;Juan Guo ,&nbsp;Jiaoqi Gao ,&nbsp;Linfeng Xie ,&nbsp;Ping Su ,&nbsp;Xiaoxin Zhai ,&nbsp;Baolong Jin ,&nbsp;Guanghong Cui ,&nbsp;Yongjin J. Zhou ,&nbsp;Luqi Huang","doi":"10.1016/j.apsb.2024.12.039","DOIUrl":null,"url":null,"abstract":"<div><div>(+)-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 <em>Saccharomyces cerevisiae</em>, and balancing cellular dephosphorylation is also helpful for efficient biosynthesis of other terpenoids since all whose biosynthesis involves the dephosphorylation procedure.</div></div>","PeriodicalId":6906,"journal":{"name":"Acta Pharmaceutica Sinica. B","volume":"15 2","pages":"Pages 1171-1182"},"PeriodicalIF":14.7000,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Acta Pharmaceutica Sinica. B","FirstCategoryId":"92","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S2211383524004969","RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q1","JCRName":"PHARMACOLOGY & PHARMACY","Score":null,"Total":0}
引用次数: 0

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.

Abstract Image

工程细胞去磷酸化促进(+)-冰片在酵母生产
(+)-冰片是中国药典中“天然冰片”的主要成分,是一种高端香料和名贵药物。植物提取不能满足对(+)-冰片日益增长的需求,而微生物合成提供了一个可持续的供应途径。然而,与其他单萜相比,即使对甲羟戊酸途径进行了广泛的优化,其产量也极低。我们发现关键的挑战是龙脑二磷酸酯(BPP)复杂而不寻常的去磷酸化反应,该反应受到细胞去磷酸化过程的副反应和竞争,特别是脂质代谢,从而限制了(+)-龙脑的合成。在这里,我们通过识别、表征磷酸酶和平衡细胞去磷酸化代谢,系统地优化了去磷酸化过程。我们首次鉴定出2种内源性磷酸酶和7种外源磷酸酶,它们显著提高(+)-冰片的产量,最高可达152%。通过对BPP去磷酸化和MVA途径的优化,(+)-冰片的产量提高了33.8倍,摇瓶补料分批发酵的产量达到753 mg/L,是迄今为止文献报道的最高产量。该研究表明,在酿酒酵母中,重连接去磷酸化代谢对于(+)-冰片的高水平生产是必不可少的,并且平衡细胞去磷酸化也有助于其他萜类化合物的有效生物合成,因为所有这些萜类化合物的生物合成都涉及到去磷酸化过程。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
求助全文
约1分钟内获得全文 求助全文
来源期刊
Acta Pharmaceutica Sinica. B
Acta Pharmaceutica Sinica. B Pharmacology, Toxicology and Pharmaceutics-General Pharmacology, Toxicology and Pharmaceutics
CiteScore
22.40
自引率
5.50%
发文量
1051
审稿时长
19 weeks
期刊介绍: The Journal of the Institute of Materia Medica, Chinese Academy of Medical Sciences, and the Chinese Pharmaceutical Association oversees the peer review process for Acta Pharmaceutica Sinica. B (APSB). Published monthly in English, APSB is dedicated to disseminating significant original research articles, rapid communications, and high-quality reviews that highlight recent advances across various pharmaceutical sciences domains. These encompass pharmacology, pharmaceutics, medicinal chemistry, natural products, pharmacognosy, pharmaceutical analysis, and pharmacokinetics. A part of the Acta Pharmaceutica Sinica series, established in 1953 and indexed in prominent databases like Chemical Abstracts, Index Medicus, SciFinder Scholar, Biological Abstracts, International Pharmaceutical Abstracts, Cambridge Scientific Abstracts, and Current Bibliography on Science and Technology, APSB is sponsored by the Institute of Materia Medica, Chinese Academy of Medical Sciences, and the Chinese Pharmaceutical Association. Its production and hosting are facilitated by Elsevier B.V. This collaborative effort ensures APSB's commitment to delivering valuable contributions to the pharmaceutical sciences community.
×
引用
GB/T 7714-2015
复制
MLA
复制
APA
复制
导出至
BibTeX EndNote RefMan NoteFirst NoteExpress
×
提示
您的信息不完整,为了账户安全,请先补充。
现在去补充
×
提示
您因"违规操作"
具体请查看互助需知
我知道了
×
提示
确定
请完成安全验证×
copy
已复制链接
快去分享给好友吧!
我知道了
右上角分享
点击右上角分享
0
联系我们:info@booksci.cn Book学术提供免费学术资源搜索服务,方便国内外学者检索中英文文献。致力于提供最便捷和优质的服务体验。 Copyright © 2023 布克学术 All rights reserved.
京ICP备2023020795号-1
ghs 京公网安备 11010802042870号
Book学术文献互助
Book学术文献互助群
群 号:481959085
Book学术官方微信