电催化还原CO2耦合代谢工程改造解脂耶氏酵母实现高效异戊二烯生物合成
Electrocatalytic CO2 Reduction Coupled with Metabolic Engineering of
Yarrowia lipolytica for Efficient Isoprene Biosynthesis- 合成生物学 2025年 页码：1-15
- 作者机构：
1.绿色生物制造全国重点实验室，北京 100029
2.北京化工大学国家能源生物炼制研发中心，北京 100029
3.北京化工大学生命科学与技术学院，北京 100029
- 作者简介：
[ "兰云龙，男（1998—），硕士研究生，主要研究方向为代谢工程。E-mail：lanyunlong@guangx.picc.com.cn" ]
[ "李嵩，男（2001—），硕士研究生，主要研究方向为代谢工程。E-mail：1144067063@qq.com" ]
[ "刘艳辉，女（1979—），博士，副教授，研究方向主要集中于酶工程及代谢工程，获得多项研究成果。发表学术论文十余篇。E-mail：liuyh@mail.buct.edu.cn" ]
- 基金信息：重点新材料研发及应用国家科技重大专项(2025ZD0614900)
DOI：10.12211/2096-8280.2025-046
中图分类号： Q939.9;Q789收稿：2025-05-19，
修回：2025-09-15，
网络首发：2025-09-17，
- 稿件说明：
移动端阅览
兰云龙， 李嵩， 张伟， 常允蕴， 刘艳辉. 电催化还原CO2耦合代谢工程改造解脂耶氏酵母实现高效异戊二烯生物合成［J］. 合成生物学， 2025， 6. DOI： 10.12211/2096-8280.2025-046
LAN Yunlong， LI Song， ZHANG Wei， CHANG Yunyun， LIU Yanhui. Electrocatalytic CO2 Reduction Coupled with Metabolic Engineering of Yarrowia lipolytica for Efficient Isoprene Biosynthesis［J］. Synthetic Biology Journal， 2025， 6. DOI： 10.12211/2096-8280.2025-046
兰云龙， 李嵩， 张伟， 常允蕴， 刘艳辉. 电催化还原CO2耦合代谢工程改造解脂耶氏酵母实现高效异戊二烯生物合成［J］. 合成生物学， 2025， 6. DOI： 10.12211/2096-8280.2025-046 DOI：
LAN Yunlong， LI Song， ZHANG Wei， CHANG Yunyun， LIU Yanhui. Electrocatalytic CO2 Reduction Coupled with Metabolic Engineering of Yarrowia lipolytica for Efficient Isoprene Biosynthesis［J］. Synthetic Biology Journal， 2025， 6. DOI： 10.12211/2096-8280.2025-046 DOI：
摘要
生物合成技术因其绿色可持续特性，成为替代传统石化工艺的重要方向。本研究通过电催化还原CO₂与代谢工程改造解脂耶氏酵母相结合的策略，实现了高效异戊二烯的生物合成。首先，在解脂耶氏酵母中异源表达不同来源的异戊二烯合酶，筛选出葛根来源的异戊二烯合酶，异戊二烯产量为560 μg/L。随后，通过强化甲羟戊酸途径关键基因
erg10、erg12、erg13、erg8、erg19、thmgr、idi
及下调竞争基因
erg20
，构建工程菌株Misps13，摇瓶发酵异戊二烯滴度提高至12.23 mg/L。进一步整合电催化-微生物耦合系统，利用自组装纳米材料L-Bi-sh-H₂O和L-Cu-sh-3将CO₂高效转化为甲酸和乙酸，其法拉第效率分别为88.67%和50.14%。在Misps13中引入ACS与FDH模块，使工程菌Misps1315能够利用电催化合成的甲酸和乙酸，增强胞内乙酰辅酶A和NAD（P）H供应。通过响应曲面法优化外源甲酸和乙酸添加量，其中甲酸2.8 g/L、乙酸6.5 g/L，最终实现异戊二烯滴度32.14 mg/L，较基础菌株提升2.6倍。本研究为生物-电化学协同制造高值化合物提供了新范式。
Abstract
Isoprene is a valuable platform chemical essential for manufacturing synthetic rubber
elastomers
and specialty materials
yet its conventional production depends heavily on petroleum resources with significant carbon emissions. Here
we present an innovative bio-electrocatalytic process that synergistically combines CO
2
electroreduction with metabolically engineered
Yarrowia lipolytica
for sustainable isoprene production. We began by screening several plant-derived isoprene synthases and identified the enzyme from
Pueraria lobata
(Isps-Pu) as the most effective in this yeast host
yielding initial production of 560 μg/L. Subsequent rational metabolic engineering involved systematic overexpression of the mevalonate pathway genes
including
erg10
erg12
erg13
erg8
erg19
thmgr
and
idi
via iterative genomic integration. Crucially
competitive metabolic flux was reduced by replacing the native promoter of
erg20
with the weak promoter P
KI1
resulting in a significantly improved isoprene titer of 12.23 mg/L.To establish an efficient electrocatalytic module
we systematically developed and characterized multiple nanomaterials for CO₂ reduction. For formate production
bismuth-based electrodes (L-Bi-sh-C and L-Bi-sh-H₂O) were synthesized through distinct self-assembly and thermal treatment pathways. Comparative analysis revealed that L-Bi-sh-H
2
O
prepared via hydrothermal treatment
demonstrated superior performance with a remarkable Faradaic efficiency of 88.67% for formate at -1.8 V versus Ag/AgCl. For acetate production
we engineered a series of copper-based catalysts (L-Cu-sh-3
L-Cu-sh-3-H₂O
L-Cu-sh-EtOH
and L-Cu-sh-EtOH-H
2
O) using different solvent systems and processing methods. Among these
L-Cu-sh-3 electrode synthesized in acetonitrile/methanol/tetrahydrofuran solvent mixture exhibited optimal performance
achieving 50.14% Faradaic efficiency for acetate at -0.8 V.The integration of these electrocatalytic components with biological conversion was achieved by introducing
Saccharomyces cerevisiae
-derived acetyl-CoA synthase and formate dehydrogenase into the engineered yeast
enabling efficient assimilation of electro-generated formate and acetate to enhance intracellular acetyl-CoA and NADPH pools. Through response surface methodology optimization
we determined optimal concentrations of formate (2.8 g/L) and acetate (6.5 g/L)
leading to a final isoprene titer of 32.14 mg/L—representing a 2.6-fold enhancement over the baseline strain. This integrated approach not only demonstrates a carbon-negative strategy for isoprene biosynthesis but also establishes a versatile platform for producing acetyl-CoA-derived chemicals from CO₂.
2
关键词
Keywords
references
CHATZIVASILEIOU A O , WARD V , EDGAR S M , et al . Two-step pathway for isoprenoid synthesis [J ] . Proceedings of the National Academy of Sciences , 2019 , 116 ( 2 ): 506 - 11 .
CUI L , XIAO Y , HU W , et al . Enhanced dataset of global marine isoprene emissions from biogenic and photochemical processes for the period 2001–2020 [J ] . Earth System Science Data , 2023 , 15 ( 12 ): 5403 - 25 .
SANADZE G . Biogenic isoprene (a review) [J ] . Russian Journal of Plant Physiology , 2004 , 51 : 729 - 41 .
KUMAR V , JOHNSON B P , DIMAS D A , et al . Novel Homologs of Isopentenyl Phosphate Kinase Reveal Class-Wide Substrate Flexibility [J ] . ChemCatChem , 2021 , 13 ( 17 ): 3781 - 8 .
WANG C-H , HOU J , DENG H-K , et al . Microbial production of mevalonate [J ] . Journal of Biotechnology , 2023 , 370 : 1 - 11 .
GASTALDO C , LIPKO A , MOTSCH E , et al . Biosynthesis of isoprene units in Euphorbia lathyris Laticifers vs. other tissues: MVA and MEP pathways, compartmentation and putative endophytic fungi contribution [J ] . Molecules , 2019 , 24 ( 23 ): 4322 .
KIM J , BAIDOO E E , AMER B , et al . Engineering Saccharomyces cerevisiae for isoprenol production [J ] . Metabolic Engineering , 2021 , 64 : 154 - 66 .
ZHENG Y , LIU Q , LI L , et al . Metabolic engineering of Escherichia coli for high-specificity production of isoprenol and prenol as next generation of biofuels [J ] . Biotechnology for biofuels , 2013 , 6 : 1 - 13 .
YANG J , NIE Q , LIU H , et al . A novel MVA-mediated pathway for isoprene production in engineered E. coli [J ] . BMC biotechnology , 2016 , 16 : 1 - 9 .
YANG C , GAO X , JIANG Y , et al . Synergy between methylerythritol phosp hate pathway and mevalonate pathway for isoprene production in Escherichia coli [J ] . Metabolic Engineering , 2016 , 37 : 79 - 91 .
ZHANG X-S , LI S , CAI H-X , et al . A DFT study of carbon dioxide reduction catalyzed by group 3 metal complexes of silylamides [J ] . Chemical Physics Letters , 2022 , 788 : 139291 .
PENG Z , DENG N , LI X , et al . Controllable Preparation, Working Mechanisms, and Actual Application of Various One-Dimensional Nanomaterials as Catalysts for CO2RR: A Review [J ] . Industrial & Engineering Chemistry Research , 2023 , 62 ( 50 ): 21511 - 35 .
YANG S , JIANG M , ZHANG W , et al . In situ structure refactoring of bismuth nanoflowers for highly selective electrochemical reduction of CO2 to formate [J ] . Advanced Functional Materials , 2023 , 33 ( 37 ): 2301984 .
QIU X F , HUANG J R , YU C , et al . A stable and conductive covalent organic framework with isolated active sites for highly selective electroreduction of carbon dioxide to acetate [J ] . Angewandte Chemie , 2022 , 134 ( 36 ): e202206470 .
LI H , OPGENORTH P H , WERNICK D G , et al . Integrated electromicrobial conversion of CO2 to higher alcohols [J ] . Science , 2012 , 335 ( 6076 ): 1596- .
BI H , WANG K , XU C , et al . Biofuel synthesis from carbon dioxide via a bio-electrocatalysis system [J ] . Chem Catalysis , 2023 , 3 ( 3 ).
LIU Q , BI H , WANG K , et al . Revealing the Mechanisms of Enhanced β-Farnesene Production in Yarrowia lipolytica through Metabolomics Analysis [J ] . International Journal of Molecular Sciences , 2023 , 24 ( 24 ): 17366 .
KAMINENI A , CHEN S , CHIFAMBA G , et al . Promoters for lipogenesis-specific downregulation in Yarrowia lipolytica [J ] . FEMS Yeast Research , 2020 , 20 ( 5 ): foaa035 .
DUAN G Y , LI X Q , DU Y R , et al . Efficient electrocatalytic reduction of CO 2 to CO on highly dispersed Ag nanoparticles confined by Poly(ionic liquid) [J ] . Chemical Engineering Journal , 2023 , 455 .
0
浏览量
1
下载量
0
CSCD
- 网络PDF下载
- WORD下载
- Meta-XML下载
- BibTeX下载
- Endnote下载
- RefMan 下载
- NoteExpress下载
- NoteFirst下载
关联资源
相关文章
相关作者
相关机构
- 地址：北京市东城区青年湖南街13号 邮编：100000
- 联系电话：010-64519339
- 技术支持由北京北大方正电子有限公司提供 京ICP备09064830号-19
京公网安备11010802024621 - 本系统建议在Chrome、 IE9+ 以上版本浏览器阅读本站内容，360浏览器请切换至极速模式
- Cookies帮助我们提供服务并提供个性化体验。使用本网站，即表示您同意我们使用Cookies