Small
·
2025
·
IF 12.1
Bingzhao Wu,Lianning Li,Hu Huang,Zhiyu Feng,Xinlong Zhang,Lei Zhang,Chunyu Zhao
Bottom-up construction of artificial cells is pivotal for investigating the functional and structural evolution of protocell systems. Here, a streamlined methodology is reported for constructing a protocell model that exhibits pH-adaptive responsiveness and functions as an efficient microreactor for cascade biochemical reactions. These protocells are engineered through liquid–liquid phase separation (LLPS) of poly(diallyldimethylammonium chloride) (PDDA) and adenosine triphosphate (ATP), yielding coacervate microdroplets that serve as nucleation sites for spontaneous self-assembly of phospholipid semipermeable membranes. By tuning the pH environment of the membranized protocells, the hydrolytic property of ATP under acidic conditions modulates the surface charge density of coacervates, triggering elevated surface potential and altered interfacial tension, which ultimately induce secondary phase separation inside the protocells to yield dynamically reconfigurable microcompartments. Notably, introduction of small-molecule substrates triggers enzymatic reactions within the molecularly crowded lumen of the protocells, which in turn drives pH-mediated micro-compartmentalization, enabling the coupling of biocatalysis with structural dynamics. This protocell model, characterized by programmable dynamic micro-compartmentalization, offers distinct advantages for applications in cellular mimicry, synthetic protobiology, and the development of artificial biomolecular microreactors.
研究领域:
合成生物学
·
生物物理学
·
分子自组装
·
原始细胞模拟
·
酶催化级联反应
✓ 已验证
产品关联
DOI
PubMed
高影响因子
ACS Synthetic Biology
·
2018
·
IF 5.316
Zhen Yao,Pingping Zhou,Bingmei Su,Sisi Su,Lidan Ye,Hongwei Yu
Isoprene, as a versatile bulk chemical, has wide industrial applications. Here, we attempted to improve isoprene biosynthesis in Saccharomyces cerevisiae by simultaneous strengthening of precursor supply and conversion via a combination of pathway compartmentation and protein engineering. At first, a superior isoprene synthase mutant ISPSLN was created by saturation mutagenesis, leading to almost 4-fold improvement in isoprene production. Subsequent introduction of ISPSLN to strains with strengthened precursor supply in either cytoplasm or mitochondria implied an imperfect match between the synthesis and conversion of the isopentenyl pyrophosphate (IPP)/dimethylallyl diphosphate (DMAPP) pool. To reconstruct metabolic balance between the upstream and downstream flux, additional copies of diphosphomevalonate decarboxylase gene (MVD1) and isopentenyl-diphosphate delta-isomerase gene (IDI1) were introduced into the cytoplasmic and mitochondrial engineered strains. Finally, the diploid strain created by mating the above haploid strains produced 11.9 g/L of isoprene, the highest ever reported in eukaryotic cells.
研究领域:
生物技术
·
代谢工程
·
合成生物学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
ACS Applied Polymer Materials
·
2024
·
IF 4.4
Meishuang Qiu,Yan Peng,Yizhuo Lu,Xiaorong Xiong,Tianyou Chen,Zushun Xu
The coimmobilization of enzymes and metal nanoparticles (MNPs) is an effective strategy for improving the efficiency of chemoenzymatic cascades. Cross-linked enzyme aggregates (CLEAs), as catalytic scaffolds, have been utilized as scaffolds for coimmobilization, leading to relatively high volumetric and space–time yields. However, it is still a challenge to achieve compartmentalization of enzymes and MNPs in CLEAs, likely due to the poor affinity between cross-linkers and MNPs. To overcome this shortcoming, here we have developed a void-adaptive cross-linking strategy for the preparation of hyperbranched polymer-cross-linked enzyme aggregates (HPCLEAs) that could be used as scaffolds for the immobilization of MNPs. Hyperbranched polymers were in situ-synthesized and could void-adaptively cross-link enzyme aggregates. They could also complex with metal precursors and then template the formation of MNPs via the in situ reduction of metal precursors, leading to the compartmentalization of several combinations of lipase and glucose oxidase (GOx) with MNPs, including Pd, Pt, Au, PdPt, AuPd, and AuPt. The physicochemical and catalytic properties of these chemoenzymatic catalysts were systematically studied to gain a deeper insight into structure–activity relationships. This strategy is promising for the development of more efficient chemoenzymatic cascades and can be extended for the compartmentalized coimmobilization of other enzymes and MNPs.
研究领域:
生物技术
·
催化
·
材料科学
·
纳米技术
✓ 已验证
产品关联
DOI
高影响因子
BIORESOURCE TECHNOLOGY
·
2025
·
IF 9
Weiwei Ge,Huihui Pai,Jiale Zhang,Chuanbo Zhang,Wenyu Lu
Miltiradiene serves as a pivotal precursor for the synthesis of numerous abietane-type diterpenes with important pharmacological activities. The endogenous mevalonate (MVA) pathway is tightly regulated in Saccharomyces cerevisiae , which limits the availability of precursors for the heterologous production of miltiradiene. In this study, the orthogonal isopentenol utilization pathway (IUP) was constructed and investigated for its adaptability with mitochondria and peroxisomes in S. cerevisiae for the synthesis of miltiradiene. Compartments combinatorial engineering was used to enhance precursor supply and miltiradiene synthesis, thereby elevating the production of miltiradiene to 146.1 mg/L in S. cerevisiae . Furthermore, an artificial multifunctional enzyme, tSmCPS-tSmKSL-PvPT, was constructed by mimicking the natural multifunctional enzyme to enhance the biosynthesis of miltiradiene in S. cerevisiae strain PCM-MT1, which is capable of producing 414.4 mg/L miltiradiene. Finally, the titer of miltiradiene was increased to 1.02 g/L by fed-batch fermentation in a 5 L bioreactor. This study broadens the application of the IUP in S. cerevisiae by integrating compartmentalization and artificial multifunctional enzymes for the synthesis of diterpenes.
研究领域:
生物技术
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代谢工程
·
合成生物学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
·
2024
·
IF 7.7
Hai Chen,Xueer Han,Xiaoyi Tan,Mengji Hu,Hongjie Dai,Yuhao Zhang
The construction of stable dual-compartmental structures is crucial for co-encapsulation and delivery of food nutrients. Herein, we explored the self-assembly beheavior and mechanism of ferritin (Fn) and aminated gelatin (AGEL), aiming to construct stable Fn/AGEL dual-compartment emulsions. Interestingly, as the AGEL concentration increased, the particle size of Fn/AGEL assemblies inversely decreased from micro- scale to nano-scale. Furthermore, the mixing dropwise manner significantly affects the size distribution of Fn/AGEL assemblies. Multiple analysis revealed that electrostatic attractions were the main chemical force driving their assembly, and both ratio and dropwise manner seem to affect the initial Fn/AGEL nucleus, ultimately altering their properties and structures. Besides, Fn/AGEL particles exhibit a reversible assembly and disassembly beheavior. Finally, stable dual-compartment emulsions with superior properties are constructed with dropwise manner of adding Fn to AGEL solution at 1:1 ratio. Overall, these findings can facilitate the development and application of ferritin-based dual-compartment emulsions.
研究领域:
食品科学
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纳米技术
·
胶体化学
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材料科学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Small
·
2022
·Vol. 19
·期 10
·2206437
·
IF 15.153
Mengqing Chen,Guoliang Liu,Ming Zhang,Yanyan Li,Xinlin Hong,Hengquan Yang
The desire for exploration of cellular functional mechanisms has substantially increased the rapid development of artificial cells. However, the construction of synthetic cells with high organizational complexity remains challenging due to the lack of facile approaches ensuring dynamic multi-compartments of cytoplasm and stability of membranes in protocells. Herein, a stable coacervate-in-Pickering emulsion protocell model comprising a membraneless coacervate phase formed by poly- l -lysine (PLys) and adenosine triphosphate (ATP) encapsulated in Pickering emulsion is put forward only through simple one-step emulsification. The dynamic distribution of intracellular components (coacervates in this protocell model) can be manipulated by changes in temperature or pH. This coacervate-in-Pickering emulsion protocell system exhibits repeatable cycle stability in response to external stimuli (at least 24 cycles for temperature and 3 cycles for pH). By encapsulating antagonistic enzymes into coacervates, glucose oxidase (GOx) and urease as an example, the control of local enzyme concentration is achieved by introducing glucose and urea to adjust the pH value in Pickering emulsion droplets. This hybrid protocell model with programmatically dynamic microcompartmentation and sufficient stability is expected to be further studied and applied in cellular biology, facilitating the development of lifelike systems with potential in practical applications.
研究领域:
细胞生物学
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合成生物学
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材料科学
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生物技术
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Nature Communications
·
2025
·Vol. 16
·期 1
·1-15
·
IF 15.7
Zhao Ke,Tan Liang-Xiao,Gao Ning,Sun Jian-Ke
Combining the chemo- and biocatalytic sites within an integrated catalyst to orchestrate complex, multi-step reactions is highly desirable yet remains a significant challenge. Here, we introduce an ionic organic cage platform for such a chemoenzymatic catalyst, achieved by electrostatic complexation of cationic molecular cage-encapsulated Pd clusters (Pd@C-Cage + ) and anionic Candida antarctica lipase B. The spatial compartmentalization provided by the cage scaffold averts undesirable coordination interactions between metal and enzyme, while also facilitating substrate channelling between dual active sites in the one-pot tandem dynamic kinetic resolution of amines, resulting in 2.1-2.7 folds enhancement in product yield within the same reaction time, compared to the physical mixture of individual analogues, and even one order of magnitude higher than the mixture of commercial immobilized lipase Novozym 435 and Pd/C. Additionally, the well-defined pore aperture and charged cage skeleton enables precise microenvironment engineering of confined metal sites, providing stringent site/shape selectivity towards substrates featuring different substituents and sizes. This platform is further demonstrated by integrating other metal clusters (e.g., Ru) and enzymes (e.g., Candida antarctica lipase A, Thermomyces lanuginosus lipase, and Glucose oxidase) for a variety of chemoenzymatic reactions, with 2.1–5.3 folds enhancement compared to the physical mixture of individual analogues.
研究领域:
催化化学
·
有机化学
·
生物无机化学
·
酶工程
·
纳米技术
·
材料科学
·
化学生物学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
CHEMOSPHERE
·
2024
·
IF 8.1
Yue Teng,Yan Yang,Zhenjun Wang,Wenjie Guan,Yutong Liu,Hongyan Yu,Luyi Zou
Aspergillus was found to be a vital hyperaccumulation species for heavy metal removal with admirable tolerance capacity. But the potential tolerance mechanism has not been completely studied. This study quantified the amounts of total cadmium (Cd), Cd 2+ , glutathione (GSH), and reactive oxygen species (ROS) in the protoplasts and vacuoles of mycelium. We modulated GSH synthesis using buthionine sulfoximine (BSO) and 2-oxothiazolidine-4-carboxylic acid (OTC) to investigate the subcellular regulatory mechanisms of GSH in the accumulation of Cd. The results confirmed that GSH plays a crucial role in vacuolar compartmentalization under Cd stress. GSH and GSSG as a redox buffer to keep the cellular redox state in balance and GSH as a metal chelating agent to reduce toxicity. When regulating the decreased GSH content with BSO, and increased GSH content with OTC, the system of Cd-GSH-ROS can change accordingly, this also supported that vacuolar compartmentalization is a detoxification strategy that can modulate the transport and storage of substances inside and outside the vacuole reasonably. Interestingly, GSH tended to be distributed in the cytoplasm, the battleground of redox takes place in the cytoplasm but not in the vacuole. These finding potentially has implications for the understanding of tolerance behavior and detoxification mechanisms of cells. In the future bioremediation of Cd in soil, the efficiency of soil remediation can be improved by developing organisms with high GSH production capacity.
研究领域:
微生物学
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环境科学
·
生物修复
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产品关联
DOI
PubMed
高影响因子
Nature Communications
·
2023
·Vol. 14
·期 1
·1-14
·
IF 16.6
Chen Feipeng,Li Xiufeng,Yu Yafeng,Li Qingchuan,Lin Haisong,Xu Lizhi,Shum Ho Cheung
Engineering heterogeneous hydrogels with distinct phases at various lengths, which resemble biological tissues with high complexity, remains challenging by existing fabricating techniques that require complicated procedures and are often only applicable at bulk scales. Here, inspired by ubiquitous phase separation phenomena in biology, we present a one-step fabrication method based on aqueous phase separation to construct two-aqueous-phase gels that comprise multiple phases with distinct physicochemical properties. The gels fabricated by this approach exhibit enhanced interfacial mechanics compared with their counterparts obtained from conventional layer-by-layer methods. Moreover, two-aqueous-phase gels with programmable structures and tunable physicochemical properties can be conveniently constructed by adjusting the polymer constituents, gelation conditions, and combining different fabrication techniques, such as 3D-printing. The versatility of our approach is demonstrated by mimicking the key features of several biological architectures at different lengths: macroscale muscle-tendon connections; mesoscale cell patterning; microscale molecular compartmentalization. The present work advances the fabrication approach for designing heterogeneous multifunctional materials for various technological and biomedical applications.
研究领域:
材料科学
·
生物医学工程
·
组织工程
·
高分子化学
·
生物材料
✓ 已验证
DOI
PubMed
高影响因子
MACROMOLECULES
·
2025
·
IF 5.2
Xiyu Wang,Qing Zheng,Xinhua Lu,Yuanli Cai
Biomimetic synthesis represents a cutting-edge topic in chemistry/materials science. Herein, we demonstrate poly(ethylene glycol) (PEG) short side-chain-assisted monomer complex coacervation and reaction-induced polyion complex compartmentalization that lead to oxygen-tolerant polymerization-induced electrostatic self-assembly (PIESA). This is achieved by the one-pot synthesis of a PEGylated anionic polyelectrolyte and heterogeneous iterative polymerization of a cationic monomer under ecofriendly ambient, in-air aqueous photo-RAFT conditions. Simultaneous reversible all-segment-participating ternary complex coacervation and Coulombic interdomain interactions lead to coacervate nanoreactors that are capable of immediate initiation and fast reversible addition–fragmentation chain transfer reactions. Approximately 2 nm monomer complex nanoclusters act as building blocks to drive liquid–liquid phase separation. Polymerization induces hierarchical self-assembly in a droplet nucleation–fusion–fission mechanism together with PEG-crowded polyion complex compartmentalization, using nanoclusters as building blocks, mechanistically similar to liquid–liquid phase separation through supramolecular polymerization. Consequently, protein-like, one-component multicompartment coacervate nanoreactors with oxygen-tolerant well-controlled fast reactions are achieved. This work provides important implications for the efficient precise synthesis of biomimetic coacervate nanodevices of increasing complexity.
研究领域:
化学
·
材料科学
·
聚合物化学
·
纳米技术
·
仿生学
✓ 已验证
产品关联
DOI
高影响因子
Science Advances
·
2023
·Vol. 9
·期 9
·
IF 13.6
He Li,Yue Yan,Jing Chen,Ke Shi,Chuwen Song,Yanglimin Ji,Liyan Jia,Jianming Li,Yan Qiao,Yiyang Lin
Engineering artificial cellular systems capable of perceiving and transmitting external signals across membranes to activate downstream targets and coordinate protocellular responses is key to build cell-cell communications and protolife. Here, we report a synthetic photoreceptor–mediated signaling pathway with the integration of light harvesting, photo-to-chemical energy conversion, signal transmission, and amplification in synthetic cells, which ultimately resulted in protocell subcompartmentalization. Key to our design is a ruthenium-bipyridine complex that acts as a membrane-anchored photoreceptor to convert visible light into chemical information and transduce signals across the lipid membrane via flip-flop motion. By coupling receptor-mediated phototransduction with biological recognition and enzymatic cascade reactions, we further develop protocell signaling–encoded Boolean logic gates. Our results illustrate a minimal cell model to mimic the photoreceptor cells that can transduce the energy of light into intracellular responses and pave the way to modular control over the flow of information for complex metabolic and signaling pathways.
研究领域:
合成生物学
·
生物物理学
·
细胞工程
·
分子生物学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
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