光驱动CO2固定生物系统直接合成多碳化学品

doi:10.1016/S1872-2067(26)64977-7

催化学报 ›› 2026, Vol. 82: 251-265.DOI: 10.1016/S1872-2067(26)64977-7

光驱动CO₂固定生物系统直接合成多碳化学品

李莹莹¹, 张健¹, 陶雨萱, 柴甜甜, 赵春雷, 陈修来^*()

江南大学生物工程学院, 工业生物技术教育部重点实验室, 江苏无锡 214122

收稿日期:2025-07-18 接受日期:2025-10-10 出版日期:2026-03-18 发布日期:2026-03-05
通讯作者: * 电子信箱: xlchen@jiangnan.edu.cn (陈修来).
作者简介:¹共同第一作者.
基金资助:
国家自然科学基金(22378166);江苏省合成生物基础研究中心基础研究计划(BK20233003);中央高校基本科研业务费专项资金(JUSRP622001);江苏省研究生科研与实践创新项目(KYCX25_2707)

Light-powered CO₂ fixation biosystem for the direct biosynthesis of multi-carbon chemicals from CO₂

Yingying Li¹, Jian Zhang¹, Yuxuan Tao, Tiantian Chai, Chunlei Zhao, Xiulai Chen^*()

School of Biotechnology and Key Laboratory of Industrial Biotechnology of Ministry of Education, Jiangnan University, Wuxi 214122, Jiangsu, China

Received:2025-07-18 Accepted:2025-10-10 Online:2026-03-18 Published:2026-03-05
Contact: * E-mail: xlchen@jiangnan.edu.cn (X. Chen).
About author:¹ Contributed equally to this work.
Supported by:
National Natural Science Foundation of China(22378166);Basic Research Program of Jiangsu and Jiangsu Basic Research Center for Synthetic Biology(BK20233003);Fundamental Research Funds for the Central Universities(JUSRP622001);Postgraduate Research & Practice Innovation Program of Jiangsu Province(KYCX25_2707)

摘要/Abstract

摘要：

目前, 随着人口增长和产业转型的发展, 二氧化碳(CO₂)等温室气体的排放急剧增加, 对环境造成了严峻的挑战. 基于CO₂转化的可持续生物制造, 不仅有助于减轻全球对化石燃料的依赖, 而且有助于实现绿色经济. 然而, CO₂的生物转化仍受到多种因素的限制, 包括: CO₂固定途径效率不高、生物转化系统中还原力与能量再生能力不足以及产物种类单一等. 因此, 开发可持续的生物固碳系统, 实现CO₂的高效资源化利用, 有助于推动绿色生物制造的可持续发展, 已成为当前研究领域的热点方向.

本文通过将人工设计的CO₂固定途径与天然类囊体膜(TKs)相耦合, 构建了一种光驱动的CO₂固定生物(LCFB)系统, 用于将CO₂直接转化为多样化的多碳化学品, 包括: 醇类、醛类、有机酸和氨基酸. 首先, 通过代谢逆合成的方法, 结合乙醛脱氢酶(ADH)、丙酮酸脱羧酶(PDC)、苹果酸酶(ME)、苹果酸激酶(MTK)和苹果酰辅酶A裂合酶(MCL)的催化功能, 构建了一条固定CO₂合成乙醛酸的丙酮酸脱羧酶/苹果酸酶(PM)循环. PM循环可以将两分子CO₂转化为一分子乙醛酸, 同时消耗两分子NADH和一分子ATP. 通过路径关键酶的挖掘与筛选, 结合路径模块化重构与优化, 并利用¹³C同位素示踪技术证明了PM循环的有效性. 其次, 为满足PM循环对能量和还原力的需求, 构建了一种基于天然TKs的光驱动能量和还原力再生引擎. 通过分离、筛选和对比不同植物来源的TKs, 发现菠菜的TKs具有最佳的能量和还原力再生性能, ATP和NADPH的再生速率分别为1.87和2.5 μmol L^-1 min^-1 μg^-1 Chl. 因此, 基于PM循环和菠菜TKs, 组装构建了LCFB系统, 用于光驱动CO₂固定合成乙醛酸, CO₂固定速率达到了2.37 nmol min^-1 mg^-1蛋白. 最后, 为了拓展LCFB系统的终端产物种类, 进一步设计与重编程了LCFB系统. 通过将LCFB系统分别与C2, C3和C4回补途径相结合, 构建了光驱动的PM-C2, PM-C3和PM-C4途径, 并以“即插即用”的模块化策略, 成功实现了利用CO₂直接生物合成多样化的多碳化学品, 包括: 醇类(如: 乙醇酸)、醛类(如: 乙醇醛和酒石酸半醛)、有机酸(如: 乙醛酸、丙酮酸、甘油酸、草酰乙酸和苹果酸)以及氨基酸(如: 甘氨酸和丝氨酸).

综上, 本文通过构建可编程的光驱动LCFB系统, 成功实现了利用CO₂直接生物合成多样化的多碳化学品, 不仅为构建碳负性多功能生物合成平台提供了新的思路, 而且为未来拓展光合系统的应用范围奠定了基础, 对推动可持续生物制造的发展也具有重要意义.

关键词: 合成光合作用, 光能, CO₂固定, 生物制造, 多碳化学品

Abstract:

Sustainable biomanufacturing depending on CO₂ conversion contributes to mitigate global dependence on fossil fuels and accelerate a future green economy. However, CO₂ conversion is limited by inefficient CO₂ fixation pathways, deficient capacity for regenerating reducing power and energy, and narrow product scope. Here, a light-powered CO₂ fixation biosystem (LCFB) was developed by coupling a new-to-nature Pyruvate decarboxylase/Malic enzyme (PM) cycle for CO₂ fixation with natural thylakoid membranes for regenerating energy and reducing power. This synthetic LCFB was able to convert CO₂ to organic molecules at a rate of 2.37 nmol min^-1 mg^-1 proteins comparable to that of natural CO₂ fixation system. Further, LCFB was programmed to extend the length of carbon chain for outputting multi-carbon chemicals by designing and constructing multiple anaplerosis CO₂ fixation pathways in a plug-and-play fashion. Finally, the programmed LCFB powered by light facilitated the direct biosynthesis of multi-carbon chemicals from CO₂, such as alcohols, aldehydes, organic acids and amino acids. This study presented here brings a potential avenue for developing a carbon-negative versatile platform to widen the applicability of LCFB and advancing sustainable biomanufacturing in the future.

Key words: Synthetic photosynthesis, Light-powered, CO₂ fixation, Biomanufacturing, Multi-carbon chemicals

李莹莹, 张健, 陶雨萱, 柴甜甜, 赵春雷, 陈修来. 光驱动CO₂固定生物系统直接合成多碳化学品[J]. 催化学报, 2026, 82: 251-265.

Yingying Li, Jian Zhang, Yuxuan Tao, Tiantian Chai, Chunlei Zhao, Xiulai Chen. Light-powered CO₂ fixation biosystem for the direct biosynthesis of multi-carbon chemicals from CO₂[J]. Chinese Journal of Catalysis, 2026, 82: 251-265.

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链接本文: https://www.cjcatal.com/CN/10.1016/S1872-2067(26)64977-7

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图/表 6

Fig. 1. Designing LCFB. LCFB consisted of the PM cycle and TKs. Acetaldehyde dehydrogenase (ADH); pyruvate decarboxylase (PDC); malic enzyme (ME), malate thiokinase (MTK); malyl-CoA lyase (MCL); nicotinamide nucleotide transhydrogenase (NNT); photosystem I (PS I); photosystem II (PS II); cytochrome (Cyt); plastoquinone (PQ); plastocyanin (PC); ferredoxin (Fd); ferredoxin NADP+ reductase (FNR).

Fig. 2. Constructing the PM cycle. (A) Schematic representation of the PM cycle for the conversion of CO2. Input module was indicated in red and output module was indicated in blue. acetaldehyde dehydrogenase (ADH); pyruvate decarboxylase (PDC); malic enzyme (ME), malate thiokinase (MTK); malyl-CoA lyase (MCL). (B) Michaelis-Menten kinetics of Acetobacter pasteurianus PDC (ApPDC) for acetaldehyde. (C) Michaelis-Menten kinetics of Clostridium acetobutylicum ME (CcME) for pyruvate. (D) Pyruvate production from acetaldehyde with different PDCs. ApPDC, PDC from Acetobacter pasteurianus, ScPDC, PDC from Saccharomyces cerevisiae, CaPDC, PDC from Clostridium acetobutylicum, ZmPDC, PDC from Zymomonas mobilis. (E) The validation of the input module. EcADH, ADH from Escherichia coli. (F) The validation of the output module. CcME, ME from Clostridium acetobutylicum; McMTK, MTK from Methylococcus capsulatus; MeMCL, MCL from Methylobacterium extorquens. (G) The validation of the PM cycle. (H) The production of glyoxylate in the different PM versions. PM-0: the initial PM cycle before optimization. PM-1: PM-0 with the optimal concentration of enzymes. PM-2: PM-1 with the optimal reaction time.

Fig. 3. Developing LCFB by coupling the PM cycle with TKs. (A) Schematic representation of LCFB for the conversion of CO2 to glyoxylate by coupling the PM cycle with TKs. (B) Steady-state absorption spectrum of TKs isolated from leek, coriander, spinach, kale and lettuce. The absorbance at 470 nm (pale blue thick vertical lines) and 650 nm (salmon pink thick vertical lines) corresponds to Chlb, the absorbance at 495 nm (pale pink thick vertical lines) is from the electron transitions of carotenoids, and the absorbance at 440 nm (yellow thick vertical lines) and 680 nm (lavender thick vertical lines) corresponds to Chla. Chla: chlorophyll a, Chlb: chlorophyll b, Cars: carotenoids. (C) Regulation of ATP production by TKs through light-dark cycles. (D) Regulation of NADPH production by TKs through light-dark cycles. (E) Production of AcCoA and glycolate with TKs. AcCoA, acetyl-CoA. Pink represents the effect of direct ATP addition and TKs-based ATP regeneration on the MTK-catalyzed reaction, respectively; blue represents the effect of direct NADPH addition and TKs-based NADPH regeneration on the GOR-catalyzed reaction, respectively. (F) Synthesis of glyoxylate in LCFBs.

Fig. 4. Programming LCFB to produce C2 compounds. (A) Schematic overview of the light-powered PM-C2 pathway. Glycolate oxidoreductase (GOR), glycolaldehyde dehydrogenase (GDH), acetyl-phosphate synthase (ACPS) and phosphate acetyltransferase (PTA). (B) The production of glycolate by ScGOR (GOR from Saccharomyces cerevisiae), AaGOR (GOR from Acetobacter aceti), NcGOR (GOR from Neurospora crassa). (C) The production of glycolaldehyde by EcGDH (GDH from E. coli), PfGDH (GDH from Pseudomonas fluorescens). (D) The verification of acetyl-CoA with SmACPS and EcPTA by LC-MS. SmACPS, ACPS from Saccharomonospora marina; EcPTA, PTA from E. coli. (E) The validation of the C2 anaplerosis module. (F) The validation of the light-powered PM-C2 pathway. All products were detected in the same reaction mixture containing the light-powered PM-C2 pathway. AcCoA, acetyl-CoA.

Fig. 5. Programming LCFB to produce C3 compounds. (A) Schematic representation of the light-powered PM-C3 pathway. Glyoxylate-aspartate aminotransferase (BhcA) or glyoxylate-alanine aminotransferase (AGX), serine aldolase (LTA) and serine dehydrase (SDA). (B) The production of pyruvate by CnSDA. (C) The production of pyruvate by PaLTA (LTA from Pseudomonas aeruginosa) and CnSDA. (D) The validation of the C3 anaplerosis module with PdBhcA (BhcA from Paracoccus denitrificans) or ScAGX (AGX from S. cerevicae). (E) The validation of the light-powered PM-C3 pathway. All products were detected in the same reaction mixture containing the light-powered PM-C3 pathway.

Fig. 6. Programming LCFB to produce C4 compounds. (A) Schematic overview of the light-powered PM-C4 pathway. Glyoxylate carboligase (GCL), tartronate semialdehyde reductase (TSR), glycerate kinase (GK), enolase (ENO), phosphoenolpyruvate carboxykinase (PCK) and malate dehydrogenase (MDH). (B) Michaelis-Menten kinetics of EcGCL (GCL from E. coli) for glyoxylate. (C) Michaelis-Menten kinetics of AsPCK (PCK from Actinobacillus succinogenes) for PEP (phosphoenolpyruvate). (D) The production of glycerate by GCL and EcTSR in the submodule I. EcGCL, GCL from E. coli; PpGCL, GCL from Pseudomonas putidis. (E) The production of PEP by EcGK (GK from E. coli) and EcENO (ENO from E. coli) in the submodule II. (F) The production of malate by AsPCK and EcMDH (MDH from E. coli) in the submodule III. (G) The validation of the C4 anaplerosis module. (H) The validation of the light-powered PM-C4 pathway. All products were detected in the same reaction mixture containing the light-powered PM-C4 pathway. PEP, phosphoenolpyruvate; OAA, oxaloacetate.

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光驱动CO₂固定生物系统直接合成多碳化学品

Light-powered CO₂ fixation biosystem for the direct biosynthesis of multi-carbon chemicals from CO₂

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