Artificial cells capable of mimicking metabolism represent a rapidly evolving frontier in synthetic biology. These systems integrate enzymes to reconstruct essential metabolic pathways, enabling the study of cellular processes in simplified yet controllable environments. This review provides a comprehensive overview of the advantages and limitations of various artificial cells for metabolic mimicry based on their physical properties. The major strategies for energy generation, including organelle encapsulation, photosynthetic phosphorylation, and nanomaterial-assisted ATP synthesis, are summarized. Anabolic processes such as carbon fixation, lipid biosynthesis, and protein expression are discussed in detail, along with representative examples of catabolic pathways involved in carbon and nitrogen metabolism. We highlight the emerging applications of metabolically functional artificial cells in biosensing and disease diagnosis. By bridging the fundamental principles and practical applications, this review aims to provide valuable insights into the design and deployment of artificial metabolic systems, paving the way for next-generation synthetic biological tools.

Phosphorus (P) is an essential macronutrient required for various vital processes in crop growth and development, including signal transduction, CO2 fixation, and photosynthetic phosphorylation. Phosphate transporters (PHTs) in plants play critical roles in the uptake, distribution, and internal transport of Phosphate (Pi). Among these transporters, the PHT4 family is widely distributed across plant species; however, the specific functions of many members within this family remain to be fully elucidated. This study focuses on unraveling the function of OsPHT4;4 in Pi utilization and photoprotection. The findings demonstrate that OsPHT4;4 acts as a low-affinity Pi transporter localized to the chloroplast membrane and reveal predominant expression of OsPHT4;4 in leaves, with peak expression during tillering and clear induction by light, exhibiting circadian rhythmicity. The ospht4;4 mutants display stunted growth. Transcriptomic analysis comparing ospht4;4 mutants and wild-types (WT) identified 1482 differentially expressed genes (DEGs), including 729 upregulated genes and 753 downregulated genes. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis reveals enrichment DEGs related to photosynthesis-antenna proteins, carbohydrate metabolism, and phenylpropanoid biosynthesis. These findings suggest that OsPHT4;4 plays crucial roles not only in photosynthesis but also in plant defense as an integral component involved in Pi metabolism.

A protet-based model for oxidative and photosynthetic phosphorylation

Two-stage (e.g. light-dark) phosphorylation experiments showed that there is a stored ‘high-energy’ intermediate linking electron transport and phosphorylation. Large, artificial electrochemical proton gradients (protonmotive forces or pmfs) can also drive phosphorylation, a fact seen as strongly supportive of the chemiosmotic coupling hypothesis that a pmf is the ‘high-energy’ intermediate. However, in such experiments there is an experimental threshold (pmf >170 mV, equivalent to ΔpH ∼2.8) below which no phosphorylation is in fact observed, and 220 mV are required to recreate in vivo rates. This leads to the correct question, which is then whether those values of the pmf generated by electron transport are large enough. Even the lower ones as required for any phosphorylation (leave alone those required to explain in vivo rates) are below the threshold [1, 2], whether measured directly with microelectrodes or via the use of membrane-permeant ions and/or acids/bases (which are always transporter substrates [3], so all such measurements are in fact artefactual). The single case that seemed large enough (220 mV) is now admitted to be a diffusion potential artefact [4]. Many other observables (inadequate bulk H+ in ‘O2-pulse’-type experiments, alkaliphilic bacteria, dual-inhibitor titrations, uncoupler-binding proteins, etc.) are consistent with the view that values of the pmf, and especially of Δψ, are actually very low. A protet-based charge separation model [2], a protonic version analogous to how energy may be stored in devices called electrets, provides a high-energy intermediate that can explain the entire literature, including the very striking demonstration [5] that close proximity is required between electron transport and ATP synthase complexes for energy coupling between them to allow phosphorylation to occur. A chief purpose of this article is thus to summarise the extensive and self-consistent literature, much of which is of some antiquity and rarely considered by modern researchers, despite its clear message of the inadequacy of chemiosmotic coupling to explain these phenomena.

Effects of polystyrene microplastic composite with florfenicol on photosynthetic carbon assimilation of rice (Oryza sativa L.) seedlings: Light reactions, carbon reactions, and molecular metabolism

Editorial

Frederick Robert (Bob) Whatley was a plant biochemist who made fundamental discoveries that form the basis of our present textbook understanding of photosynthesis, the process by which green plants harness the energy of sunlight to incorporate atmospheric carbon dioxide, and upon which all life on Earth depends. Bob was born in Wiltshire and graduated from the University of Cambridge with a first class degree in biochemistry and a PhD supervised by Robin Hill FRS. He brought biochemical expertise and insight to a small team under the leadership of Daniel Arnon at the University of California at Berkeley, which, in the decade from 1953, discovered that chloroplasts were capable of conserving light energy as adenosine triphosphate by what the Berkeley group named photosynthetic phosphorylation. The Berkeley group also defined the role of the enzyme cofactor NADP + . By demonstrating that isolated chloroplasts were capable of the complete process of light-driven carbon dioxide fixation, they revolutionized the contemporary conception of the process, and provided a solid basis for subsequent research. In the 1960s Bob Whatley contributed to identifying the important role of the key protein, ferredoxin, in photosynthesis, and he established an important centre for photosynthesis research at King's College London. In the 1970s and 1980s as Sherardian professor of botany at Oxford he advanced several areas of biochemical research. Bob Whatley was a meticulous experimenter and clear thinker. He had a gentle manner and, for all his scientific achievements, Bob possessed an inherent modesty.

The effects of superoxide dismutase (SOD) inhibitors, diethyldithiocarbamate (DDC), triethylenetetramine (trien), and their combination with glucose on cells of the epidermis from pea leaves of different age (rapidly growing young leaves and slowly growing old leaves) was investigated. DDC and trien caused death of the guard cells as determined by destruction of their nuclei. Glucose did not affect destruction of the nuclei induced by SOD inhibitors in the cells from old leaves, but intensified it in the cells from young leaves. 2-Deoxyglucose, an inhibitor of glycolysis, and propyl gallate, SOD-mimic and antioxidant, suppressed destruction of the nuclei that was caused by SOD inhibitors and glucose in cells of the epidermis from the young, but not from the old leaves. Glucose and trien stimulated, and propyl gallate reduced generation of reactive oxygen species (ROS) in the pea epidermis as determined by the fluorescence of 2',7'-dichlorofluorescein (DCF). Carbonyl cyanide m-chlorophenylhydrazone (CCCP), a protonophoric uncoupler of oxidative and photosynthetic phosphorylation, suppressed the DCF fluorescence in the guard cells. Treatment of the cells with CCCP followed by its removal with washing increased destruction of the nuclei caused by SOD inhibitors and glucose. In young leaves, CCCP was less effective than in old ones. The findings demonstrate the effects of SOD inhibitors and glucose on the cell death and generation of ROS and could indicate glycolysis-dependent ROS production.

Significant strides toward producing biochemical fuels have been achieved by mimicking natural oxidative and photosynthetic phosphorylation. Here, different from these strategies, we explore boric acid as a fuel for tuneable synthesis of energy-storing molecules in a cell-like supramolecular architecture. Specifically, a proton locked in boric acid is released in a modulated fashion by the choice of polyols. As a consequence, controlled proton gradients across the lipid membrane are established to drive ATP synthase embedded in the biomimetic architecture, which facilitates tuneable ATP production. This strategy paves a unique route to achieve highly efficient bioenergy conversion, holding broad applications in synthesis and devices that require biochemical fuels.

Abstract Significant strides toward producing biochemical fuels have been achieved by mimicking natural oxidative and photosynthetic phosphorylation. Here, different from these strategies, we explore boric acid as a fuel for tuneable synthesis of energy‐storing molecules in a cell‐like supramolecular architecture. Specifically, a proton locked in boric acid is released in a modulated fashion by the choice of polyols. As a consequence, controlled proton gradients across the lipid membrane are established to drive ATP synthase embedded in the biomimetic architecture, which facilitates tuneable ATP production. This strategy paves a unique route to achieve highly efficient bioenergy conversion, holding broad applications in synthesis and devices that require biochemical fuels.

A protet-based, protonic charge transfer model of energy coupling in oxidative and photosynthetic phosphorylation.

High light causes disturbances in photosynthetic phosphorylation or damage to the photosystem II (PSII) structure or even assimilation tissues. The value of the red/ far-red ratio (R/FR) provides the plant with information on the environmental light conditions, regulating, among others, photosynthetic activity and pigment composition of the plant. The response of the photosynthetic apparatus of the sporotrophophylls and nest leaves of Platycerium bifurcatum, grown for 6 months at the low or high R/FR ratio, were studied. Later, the plants were transferred to high light (1,200 µmol quantum • m-2 • s-1). Changes in PSII photochemical activity were determined based on non-destructive methods of chlorophyll a fluorescence kinetics analysis. The measurement of radiation reflectance from the leaves allowed to determine the content of selected pigments related to the photosynthesis process and to assess changes in the Photochemical Reflectance Index. The calculation of reflectance difference and sensitivity analysis was used to identify so-called "stress-sensitive wavelengths". Plant growth at high R/FR ratio prepares photosynthetic apparatus of ferns to high light and enables more efficient conversion of absorbed photons. The increase in the amount of photoprotective compounds allows the protection against photoinhibition in the sporotrophophyll leaves that play key roles in plant nutrition and reproduction.

Photosynthesis of Microcystis aeruginosa under Electromagnetic Radiation (1.8 GHz, 40 V/m) was studied by using the proteomics. A total of 30 differentially expressed proteins, including 15 up-regulated and 15 down-regulated proteins, were obtained in this study. The differentially expressed proteins were significantly enriched in the photosynthesis pathway, in which the protein expression levels of photosystems II cytochrome b559 α subunit, cytochrome C550, PsbY, and F-type ATP synthase (a, b) decreased. Our results indicated that electromagnetic radiation altered the photosynthesis-related protein expression levels, and aimed at the function of photosynthetic pigments, photosystems II potential activity, photosynthetic electron transport process, and photosynthetic phosphorylation process of M. aeruginosa. Based on the above evidence, that photoreaction system may be deduced as a target of electromagnetic radiation on the photosynthesis in cyanobacteria; the photoreaction system of cyanobacteria is a hypothetical “shared target effector” that responds to light and electromagnetic radiation; moreover, electromagnetic radiation does not act on the functional proteins themselves but their expression processes.

Two-ion theory of energy coupling in ATP synthesis rectifies a fundamental flaw in the governing equations of the chemiosmotic theory

Keynote Summary: Half a century on huanglongbing: learning about the disease, trying to control it

The pathway of adenosine triphosphate (ATP) synthesis in living organisms consists of two autonomous stages; this must be taken into account during the design and analysis of chemical models of the abiogenesis of this key participant of metabolic processes. The first stage is construction of an adenine heterocycle linked to a ribose-5-phosphate molecule to yield AMP, while the following stage is the attachment of phosphoryl residues to the nucleotide molecule by macroergic phosphoanhydride bonds. Involvement of the same set of precursor molecules in both de novo biosynthesis of AMP and abiogenesis of this nucleotide is a very important issue for the analysis of metabolic pathways. Photochemical matrix systems that convert light energy into macroergic bond energy are functional prototypes of photosynthetic phosphorylation; they are of special interest for the construction of abiotic phosphorylation models. Interaction of substrates with a purely mineral matrix (montmorillonite particles) under UV irradiation resulted in the formation of ATP from ADP and orthophosphate. Micro-and nanostructures that formed upon the interaction of the mineral component (sodium polysilicate) with an abiogenic organic pigment (a flavin conjugate with a random amino acid polymer) exhibited phosphorylating activity as well when irradiated with visible light. The properties of AMP and ATP abiosynthesis models investigated are in good accordance with the current views on the environmental conditions of the ancient Earth; evident structural differences exist between these models and the biosynthetic systems in modern organisms.

JosephM. Bovewas born in Luxemburg, 1929. French citizen since 1968. Higher education: School of Agronomy and University, Paris, France (1950-1955), University of California, Berkeley (1956-1955). Doctorate on in vitro synthesis of plant viral RNA (1967). Researcher at the French Institute for Citrus and Tropical fruit Research (1959-1970) at Versailles, France. Director of research at INRA (French National Institute for Agricultural Research), at INRA campus of Bordeaux, France (1971-1975) and professor of microbiology at University of Bordeaux (1976-1997). Head of Laboratory for Cellular and Molecular Plant Biology (1974-1994). President of INRA-Bordeaux (19841994). FAO consultant for citrus diseases (1981-1993). Consultant at Fundecitrus, Sao Paulo State, Brazil, for graft-transmissible diseases of citrus (1998-2014). Research. (i) Photosynthetic phosphorylation, laboratory of Prof. D.I. Arnon, University of California at Berkeley (1956-1959). (ii) Replication of plant virus RNA, Versailles and Bordeaux (1959-1986). (iii) Discovery and study of pathogenic, phloem-restricted bacteria of citrus: Spiroplasma citri, agent of stubborn disease, Candidatus Liberibacter spp., associated with huanglongbing, Candidatus Phytoplasma aurantifolia, associated with witches’ broom disease of lime, as well as of xylem-restricted bacteria: Xylella fastidiosa, agent of citrus variegated chlorosis (1969-1999). (iv) Etiology and management of citrus diseases in Brazil: Variegated chlorosis (1989-1993), Sudden Death (19982005), Huanglongbing 2005-2014). J.M. Bove is member of the French Academy of Agriculture (1992), corresponding member of the French Academy of Science (1993), member of the Brazilian Academy of Science (2002), Fellow of the American Phytopathological Society (1994), Fellow of the International Organization of Citrus Virologists (2004). Phytoparasitica DOI 10.1007/s12600-014-0415-4

New perspectives on photophosphorylation have been offered from the standpoint of the torsional mechanism of energy transduction and ATP synthesis. New experimental data on the involvement of malate anions in ATP synthesis in an acid-base malate bath procedure has been reported on spinach chloroplast thylakoids as the model system. The data cannot be reconciled with the chemiosmotic theory but has been shown to be naturally explained by the torsional mechanism. The path of malic acid in the acid and base stages of the experiment has been traced, offering further strong support to the new paradigm. Classical observations in the field have been re-interpreted in the light of these findings. A new concept of ion translocation, energy transduction and coupling at the overall physiological level in photophosphorylation has been presented and a large number of novel experimentally testable predictions have been made and shown to arise as logical consequences of the new perspectives.

Cyanide is an apoptosis inducer in stoma guard cells from pea leaf epidermis. Unlike CN-, the uncoupler of oxidative and photosynthetic phosphorylation carbonyl cyanide m-chlorophenylhydrazone (CCCP), the combination of CCCP, 3-(3 ,4 -dichlorophenyl)-1,1-dimethylurea (DCMU), benzylhydroxamate (BH), myxothiazol, antimycin A, and a glycolysis inhibitor 2-deoxyglucose (DG) did not induce destruction of guard cell nuclei for 20 h of incubation of epidermal peels in the light. DCMU prevented the effect of CN- as a programmed cell death (PCD) inducer. CCCP, the combination of DCMU and CCCP, or the combination of DCMU, CCCP, BH, myxothiazol, antimycin A, and DG supplemented by CN- caused destruction of cell nuclei; the number of the cells lacking nuclei in this case was higher than with CN- alone. DG and CCCP caused cell destruction after longer incubation of the isolated epidermis - after 2 days and to a greater degree after 4 days. The effect of DG and CCCP was reduced by illumination. Cell destruction during long-term incubation was prevented by the combination of DG and CCCP. From data of electron microscopy, DCMU and dinitrophenyl ester of iodonitrothymol (DNP-INT) prevented apoptotic changes of the nuclear ultrastructure induced by CN-. The suppression of the destruction of the guard cell nuclei under combined action of DG and CCCP was apparently caused by switching of cell death from PCD to necrosis. Thus, the type of cell death - via apoptosis or necrosis - is controlled by the level of energy provision.

By the early 1970s, the chemiosmotic hypothesis of Peter Mitchell was widely accepted by bioenergetics researchers as the best conceptual scheme to explain how ATP is formed in oxidative and photosynthetic phosphorylation. At about the same time, however, work from a few laboratories suggested that some aspects of that elegant, relatively simple hypothesis required revision - not abandonment, but refinement to accommodate more complex movements of protons in the ATP formation mechanism than originally envisioned by Peter Mitchell. In some situations it appeared that protons were constrained to localized domains rather than always delocalized within an enclosed vesicle as envisioned by chemiosmosis. This minireview tells that story from my perspective, as one of the researchers involved in the experimental approaches that revealed more complex energy coupling proton flux patterns. Ionic conditions during isolated thylakoid storage were found to reversibly switch the [Formula: see text] gradient driving ATP formation between delocalized and localized energy coupling modes. Thylakoid accessible Ca(2+) ions proved to be the switching factor that was responding to the ionic conditions in the storage treatment. The mechanism of Ca(2+) was at least partially demystified when it was shown that the reversible switching between [Formula: see text] energy coupling modes involved Ca(2+) interactions with the 8 kDa CF(0) (the H(+) channel) subunit in a type of H(+) flux gating action. Other experiments showed that the Ca(2+) gating of H(+) flux into the lumen may be a critical regulatory factor in controlling the lumen pH and thereby help regulate the activity of the violaxanthin de-epoxidase enzyme, a key part of the chloroplast photoprotective response to over-energization (excess light) stress.