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Results for “biochemistry”

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bioRxiv2026

Makeshift: a lightweight software for accessing and analyzing NMR data and protein dynamics

Nuclear magnetic resonance (NMR) spectroscopy yields rich residue-level information on biomolecular dynamics and chemical environments, two frontiers for quantitative predictive methods in biochemistry. Decades of data are publicly archived in the Biological Magnetic Resonance Data Bank (BMRB)1, yet in practice, this information remains difficult to access and interpret at scale and within computational workflows. Here we present makeshift, an open-source Python package for accessing, curating, and analyzing NMR datasets. Users can readily retrieve and parse BMRB entries and perform essential analyses such as chemical shift re-referencing, secondary structure propensity prediction, and interpretation of relaxation datasets for dynamics. We re-implemented several widely-used NMR data calculations which were not open-source or available in Python and validated our implementations against the original implementations. By integrating data access, processing, and analysis into a single Python interface, makeshift lowers the barrier for reproducible, scalable analysis and machine learning applications using biomolecular NMR data.

biophysics↗

bioRxiv2026

Altered axonal initial segment development links circuit and Kv7 dysfunction in an Fmr1 knockout rat

Fragile X syndrome (FXS) is the leading monogenetic cause of intellectual disability and autism, yet how basic hippocampal circuit properties evolve across development for this condition remains unclear. Here, we studied CA1 pyramidal neurons in male Fmr1 knockout rats at postnatal day (P)12 - 15 and 6 - 10 weeks using ex vivo electrophysiology pharmacology, imaging and biochemistry. P12 - 15 knockout neurons showed impaired sustained firing, progressive action potential broadening and enhanced activity-dependent synaptic vesicle replenishment. These defects recovered by 6 -10 weeks. These phenotypes were linked to Kv7 channel dysfunction, as Kv7 activation altered action potential dynamics and neurotransmission in WT but not Fmr1 KO neurons. Rather than directly altering Kv7 channel function, Fmr1 knockout altered axon initial segment (AIS) development such that Kv7 became functionally inert. These findings suggest that this drives early, transient Kv7-dependent CA1 dysfunction in FXS and highlight how alterations in AIS development can have profound functional impacts.

neuroscience↗

bioRxiv2026

ORB-TXTL: cell-free expression of membrane proteins on lipid bilayer-coated beads

Membrane proteins achieve a remarkable range of cellular functions, yet their characterization at high throughputs remains difficult with standard reconstitution methods. Here, we develop On-bead Reconstitution into Bilayers via Cell-free Transcription and Translation (ORB-TXTL), a platform that uses compositionally tunable lipid bilayer-coated silica beads as scaffolds for cell-free synthesized interacting and integral membrane proteins. ORB-TXTL is fast as it just takes a few hours to integrate membrane proteins onto the beads, which can be extensively washed and seamlessly transferred between reaction buffers, to perform assays that are read out by standard laboratory equipment without tagging and sophisticated equipment. We first characterized the lipid interactions of the mechanosensitive channel MscL, then screened 169 E. coli proteins and identified a systematic dependence of membrane integration efficiency on the number of transmembrane domains. Finally, we functionally reconstituted the E. coli phospholipid synthesis pathway, demonstrating that ORB-TXTL is a tractable and cheap chassis for multi-enzyme membrane biochemistry.

synthetic biology↗

bioRxiv2026

Dark CO2 Fixation via the Ethylmalonyl-CoA Pathway Establishes Metabolic Parity: A Stoichiometric Basis for Compounding Ecosystem Shifts

A central challenge in microbial ecology is understanding how competing guilds coexist and why community structures unexpectedly drift away from theoretical steady states. While polyphosphate accumulation provides energy and redox buffering in many heterotrophs, how competing lineages occupying the same ecological niche without polyphosphate synthesis manage equivalent intracellular redox imbalances remains unposed. Using an enhanced biological phosphorus removal macrocosm, we integrated quantitative stoichiometry with metaproteomics to resolve alternative metabolic strategies that underpin cellular homeostasis in heterotrophs. We show that glycogen-accumulating organisms (GAOs) achieve baseline metabolic parity with polyphosphate-accumulating organisms (PAOs) through a parallel redox-buffering mechanism: heterotrophic CO2/HCO3- re-assimilation via the ethylmalonyl-CoA pathway. This inorganic carbon fixation couples structural carbon conservation with tight redox control, mitigating intracellular electron overflow and eliminating the long-assumed GAO bioenergetic inferiority. The bioenergetic efficiency of GAOs is further fortified by fine-tuned metabolic wiring, featuring energy-efficient high affinity acetate activation, ferredoxin-centered biochemistry, and energy-neutral polyhydroxyalkanoates (PHA) mobilization. Strikingly, minor formate co-feeding disrupted this established PAO/GAO parity. Stoichiometric simulations revealed that this formate supplementation creates an asymmetric bioenergetic niche that grants per-cycle energy gains exclusively to GAOs. Decoupled from hydraulic throughput, solids retention time control retains cells carrying accumulated intracellular inventory, translating subtle per-cycle stoichiometric edges into a multi-generational ratchet and drives a rapid community shift from PAO/GAO co-dominance to GAO dominance. By exposing limitations of traditional single-substrate and single-cycle steady-state models, our findings reveal how inorganic resource management and generational metabolic compounding govern community assembly in biomass-retaining microbial ecosystems.

bioengineering↗

bioRxiv2026

Interval-zone free-flow electrophoresis as a charge-specific dimension for native isolation of thylakoid membrane protein complexes

Native polyacrylamide gel electrophoresis is the standard first analytical step after detergent solubilization of thylakoid membranes. It separates photosystem supercomplexes by size and shape, but it forces a polydisperse mixture of protein-detergent particles through a gel of limited pore diameter. Interval-zone free-flow electrophoresis (iZE-FFE) performs that first electrophoretic step in free solution, without ampholytes or immobilines, and returns the complexes as a liquid fraction series ordered by net charge. We describe the operational cycle of iZE, show that bromophenol blue reports the charge density of {beta}-dodecylmaltoside micelles, and apply iZE to Arabidopsis thaliana thylakoid extracts from two sequential solubilization campaigns. After a two-step digitonin then digitonin/{beta}-dodecylmaltoside extraction (experiment 6874), SDS-PAGE of the iZE fractions is crowded: high-molecular mass protein-lipid assemblies of photosystem I and photosystem II release the same subunits into many wells. Native-PAGE of the same fractions resolves ATP-synthase from PSI, a free LHCII pool, anodic PSII, and cathodic PSI, including PSI with and without LHCII. A three-step extraction with a single pH working window (experiment 7154) confirms this charge order. Lowering residual detergent in consecutive extracts still produces partly solubilized high-mass assemblies, whereas a final {beta}-dodecylmaltoside step releases the C2S2M2/C2S2M/C2S2 PSII series and a single PSI band. iZE is therefore a charge-first, orthogonal dimension for native thylakoid biochemistry. Native-PAGE, not SDS-PAGE, is the proper second dimension while solubilization remains incomplete.

plant biology↗

bioRxiv2026

Four numbers, one axis: deep learning models reveal what leaf spectrum constrains about Farquhar-von Caemmerer-Berry photosynthesis

Leaf reflectance spectra are emerging as a viable substitute for gas-exchange measurements of photosynthetic capacity, with a community benchmark reporting that a spectrum accurately recovers most Farquhar-von Caemmerer-Berry (FvCB) parameters. This study re-scores the recovery under dataset-blocked, species-blocked, and leave-one-dataset-out designs, measuring the split-half reliability of each curated parameter. We constructed a convolutional encoder that maps a spectrum to the four parameters through a fixed, differentiable FvCB decoder trained on measured assimilation. A conspecific of 97.4% of held-out leaves were present in the training set, and accuracy is lost along the dataset axis but not along the species axis. Under blocked evaluation, a spectrum constrains a single capacity axis. Jmax25 retains only 17% of its recovery when Vcmax25 is held constant, and the Jmax25:Vcmax25 ratio is not predicted above a median null. The curated values of TPU25 are not reproducible, whereas those of Rday25 are well determined, but its recovery fails due to the loss. The published study measures interpolation rather than transfer, and spectra constrain less of the FvCB parameter space than assumed, including the carboxylation to electron transport balance. Routing predictions through explicit biochemistry makes identifiability measurable, although it does not improve prediction accuracy.

plant biology↗

bioRxiv2026

Centrosome maintains the integrity of and repairs mature olfactory cilia in adult Drosophila

Mechanisms of cilium assembly are well studied; however, how mature metazoan cilia maintain their structures and functions in vivo remains unknown. It is also unclear whether the centrosome-derived basal body (BB) directly contributes to ciliary homeostasis. We combined biochemistry, genetics, high-resolution subcellular imaging, and electrophysiology to investigate long-lived ciliated olfactory sensory neurons (OSNs) and their olfactory behaviour in adult Drosophila. Several centrosome assembly proteins are absent, but another subset persists through dynamic protein exchange at the BBs of mature olfactory cilia. At this ciliary base, pericentriolar material (PCM) components, e.g., {gamma}-Tubulin23C, centrosomin, and pericentrin-like protein form an interconnected network required for ciliary maintenance. Adult- and OSN-specific depletions of these proteins, particularly in combination, disrupt accumulation of the heterotrimeric kinesin-2 at the ciliary base and, subsequently, in the shaft, causing loss of ciliary tubulin, EB1, and odorant receptor co-receptor. These dysregulations cause profound loss of ciliary shaft and impair olfactory function/behaviour. The centrosomal kinases PLK1/POLO and Aurora A also work with this PCM network and are required for mature ciliary homeostasis. Remarkably, these defects in ciliary structure, function, and adult olfactory behaviour are reversible, suggesting that this centrosome-derived BB is a dynamic homeostatic epicentre that regulates trafficking and ciliary structural and compositional integrity in vivo. These findings uncover an active, centrosome-dependent, cell-autonomous mechanism for maintaining and repairing mature metazoan cilia in fully grown organs in adulthood and also provide a possible explanation for how deregulation of conserved ciliary base components could lead to progressive, late-onset cilia-related disorders.

cell biology↗

bioRxiv2026

Net conversion calculations of catabolic pathways

Motivation: The stoichiometry (or net conversion) of a catabolic pathway is an often used principle of biochemistry. It expresses the molar yield of charged energy carriers (e.g. ATP) and catabolic products (e.g. lactate) on the energy source (e.g. glucose). Product yields are engineering targets of metabolic engineering and used in microbial ecology to assess energy metabolisms of microbial species. For a single species under a single condition, the catabolic pathway is frequently assumed fixed, while there might be multiple options encoded in its genome. To find these options, the manual (heuristic) methods that have been used for decades fall short. Results: In this paper, we explain how net conversions can be calculated from reaction stoichiometries of a metabolic network and evaluated using thermodynamic information. We start with an (old) heuristic method. Next, we explain we relate the net conversions of metabolic networks to their elementary flux modes (EFMs) and show that a single EFM gives rise to a single net conversion. EFMs are mathematical objects that are computable with existing software. We use these to illustrate how all net conversions of complex (pan-)metabolic networks can be computed. We consider examples from aerobic and anaerobic microbiology. Then, we introduce the parameter {Omega}, the driving force per unit flux, which allows for the thermodynamic comparison of pathways. To calculate {Omega}, only the standard Gibbs free energy potential, i.e., {Delta} G m' of the net conversion and its corresponding EFM are required. A generic workflow (and all underlying Python code) are provided, as well as applications to perform the workflow without coding. We also provide those software packages that automate our methods. Impact: This paper serves as an illustration of how modern computational systems biology can be used to automate the calculation of net conversions in microbial ecology and metabolic engineering. We hope that this paper inspires future metabolism research using quantitative, rigorous methods.

systems biology↗

bioRxiv2026

Chromatin structure of the inactive X chromosome revealed by in situ cryo-electron tomography

The intricate organisation of chromatin in eukaryotic cells plays a central role in regulating gene transcription and other DNA-templated processes. Prior studies using light microscopy, biochemistry, and genomics have implicated histone and DNA modifications, histone variants, chromosomal proteins, non-coding RNA, and intrinsic biophysical properties of chromatin in determining chromatin organisation across multiple scales. In this study, we apply an innovative approach, direct visualisation of native chromatin structure and its modulation by different factors using cryo-electron tomography, exploiting the inactive X chromosome (Xi) in differentiating mouse embryonic stem cells as a model system of facultative heterochromatin. We show that Xi chromatin undergoes progressive compaction across scales, from individual nucleosomes to chromatin domains. We observe an enrichment of histone H1-bound chromatosomes that are distributed to form the core of Xi chromatin domains. Moreover, we demonstrate that histone deacetylation demarcates chromatin domains. Specifically, perturbations leading to histone acetylation in the Xi dissolve discrete chromatin-domain boundaries, resulting in a homogeneous chromatin distribution with a dense and uniform packing of nucleosomes. Together, these findings illuminate the multiscale organisation of native facultative heterochromatin and the contributions of histone H1 and histone acetylation to this organisation. This study opens a new avenue for investigating how chromatin modification and structure relate to gene activity in near-native cells.

molecular biology↗

bioRxiv2026

TERT interacts with TFAM to activate mitochondrial fragmentation and auto/mitophagy

Previous studies have demonstrated that Telomerase Reverse Transcriptase (TERT) is localized inside mitochondria, where it associates with mitochondrial DNA (mtDNA), thereby influencing replication and transcription. Using a combination of quantitative fluorescence microscopy and biochemistry, we demonstrate that TERT binds indirectly to mtDNA through the interaction with mitochondrial transcription factor A (TFAM) to induce mitochondrial fragmentation. These findings reveal a novel mitochondrial interaction involving TERT and highlight a role for TERT and TFAM in the regulation of auto/mitophagy. This interaction is an important factor for mitochondrial function and contributes to the remodelling of mitochondrial respiratory capacity.

cell biology↗

bioRxiv2026

Single molecule detection of Tau seeds in extracellular vesicles from Alzheimer's disease brains

Extracellular vesicles (EVs) play a critical role in the propagation of Tau pathology in Alzheimer's disease (AD). The selective identification and characterization of Tau seed-carrying EVs would help developing new diagnostic tools and therapeutic strategies. While pathogenic forms of Tau can be detected biochemically in EV-enriched fractions, only super-resolution fluorescence imaging provides both morphological and compositional information at the level of single EVs on account of their small size and the low number of target molecules. This was the object of our study. Post-mortem human brain-derived EVs from AD patients and non-demented control individuals were characterized in depth using biochemistry, transmission electron microscopy (TEM) and single molecule localization microscopy (SMLM). Dual-colour SMLM of immobilized EVs, displaying size distributions that were consistent with TEM, enabled statistically meaningful measurements of thousands of EVs in parallel. Owing to the high spatial resolution and capability of SMLM to detect sparse epitopes, we identified pathological Tau proteins in individual EVs from AD samples. Reconstructed super-resolution images of permeabilized EVs demonstrate that pathological Tau forms are frequent cargo proteins of EVs from AD brains. Our data illustrate the power of SMLM to determine the ultrastructure, molecular composition and topology of single EVs and EV populations.

neuroscience↗

bioRxiv2026

Preclinical evaluation of the biodistribution, safety, and hemocompatibility of PEG-coated iron oxide magnetic nanoparticles in a viral oncogenesis mouse model

Previously, we demonstrated the biosafety of polyethylene glycol-coated iron oxide nanoparticles (MAG@PEG) in a cellular model of viral oncogenesis, supporting their further preclinical evaluation. Herein, we evaluated the safety and biodistribution of MAG@PEG in tumor-bearing mice and assessed the influence of an external magnetic field on nanoparticle distribution. Animals were randomly assigned to three groups: G1, control (PBS); G2, MAG@PEG (10 mg/kg); and G3, MAG@PEG (10 mg/kg) with magnetic exposure for 1 h. Doses were administered intraperitoneally weekly for 1 month. Results showed that animal behavior and body weight were unaffected by MAG@PEG, while tumor growth was significantly reduced only in G2. Biodistribution studies based on tissue magnetization revealed higher magnetic iron levels in the stomach followed by the spleen, lungs, and tumors, particularly in G2. Histological analysis detected Prussian blue-positive staining mainly in the spleen and in lymphoid-associated structures in the lung, stomach and intestine, while a fraction of MAG@PEG remained within the peritoneal cavity. Serum biochemistry showed no evidence of nephrotoxicity. Furthermore, hemocompatibility studies using human blood samples demonstrated that MAG@PEG was non-hemolytic according to the ASTM F756 standard. Taken together, the biodistribution, histological, biochemical, and hemocompatibility findings provide a comprehensive preclinical assessment of the biosafety of MAG@PEG and identify the route of administration and magnetic targeting as key determinants of nanoparticle biodistribution.

pharmacology and toxicology↗

bioRxiv2026

The constitutive androstane receptor protects against metabolic dysfunction-associated steatotic liver disease through sex specific interactions with the gut microbiota

Background: The gut-liver axis is involved in Metabolic dysfunction-Associated Steatotic Liver Disease (MASLD), but the host molecular actors involved in such dialogue have not yet been fully characterized. Objective: We investigated the sex-dependent role of the Constitutive Androstane Receptor (CAR) as a hepatic sensor of microbial signals in MASLD pathophysiology. Design: Male and female Car+/+ and Car-/- mice were fed a western diet (WD) for 4, 14 or 35 weeks. We performed plasma biochemistry, liver histology, bile acid and cholesterol quantification, untargeted multi-omics approach combining liver RNA-sequencing and 16S rRNA sequencing, together with co-housing, fecal microbiota transplantation and analysis of publicly available transcriptomics data in human cohorts. Results: Short-term WD revealed CAR-dependent protection against steatosis, together with perturbations in the transcriptional signatures of multiple transcription factors involved in hepatocyte identity in males. After 14 weeks, CAR deletion increased WD-induced liver damage, inflammation and fibrosis, and disrupted bile acid, cholesterol and gut microbiota homeostasis, in males only. Co-housing and fecal microbiota transplantations revealed that a Car+/+ gut microbiota protected against WD-induced liver damage, while a Car-/- microbiota was reshaped by a Car+/+ host. No CAR-dependent phenotype was observed in females. Finally, in human cohorts, hepatic CAR expression decreased in advanced clinical stages of MASLD. Conclusion: CAR is a sex-dependent protective nuclear receptor against MASLD progression, acting both as a transcriptional regulator of hepatocyte identity and as a mediator of the host-microbiota crosstalk.

physiology↗

bioRxiv2026

Parathyroid Hormone Receptor 1 Signaling Encodes Multifaceted Metabolic Programs in AgRP Neurons

Organisms continuously adapt food intake and nutrient utilization to fluctuating energy demands, requiring the brain to accurately integrate signals coming from diverse metabolic organs and local signals into distinct physiological responses. How hypothalamic neurons discriminate these signals to independently regulate feeding behavior and peripheral metabolism remains poorly understood. In recent years, a coordinated regulation between bone physiology and the central control of energy metabolism has emerged, opening questions about potential common hormonal networks and signaling pathways between the bone and the brain. Here, we identify parathyroid hormone receptor 1 (PTH1R) signaling, a master regulator of calcium ion biochemistry and bone formation, in Agouti-related peptide (AgRP) neurons as an unexpected neuroendocrine mechanism for the control of feeding behaviourand nutrient partitioning. the two cognate endocrine and paracrine ligands, PTH and PTH-related peptide (PTHrP), selectively activat segregated intracellular kinases activation and lipophagic processes in AgRP neuron, ultimately producing convergent physiological outputs. Together, our findings identify the parathyroid hormonal system acts as a fine-tuning mechanism in specialized neuronal circuits allowing for hypothalamic adaptive responses to energy demands and systemic metabolic signals. Keywords: Parathyroid hormone-related peptide (PTHrP), hypothalamus, arcuate nucleus, agouti-related peptide neurons, energy metabolism, fasting, autophagy, free fatty acids

neuroscience↗

bioRxiv2026

Neuronal FICD shapes motivated behavior through modulation of dopamine receptor dynamics

Motivated behaviors rely on dopaminergic signaling, which in turn depends on the correct folding and trafficking of receptors through the endoplasmic reticulum (ER). Within the ER, the AMPylase enzyme FICD acts as a molecular rheostat through its modification of the chaperone BiP, dynamically tuning the organelle's responsiveness to folding perturbations and cellular stress. While the biochemistry of FICD is well-established, its role in complex animal behavior remains poorly understood. Here, we identify FICD as a conserved contributor to the generation of motivated behavior. In Drosophila, pan-neuronal knockdown of dFic significantly reduces feeding, spontaneous locomotion, and reward-seeking behavior. These alterations occur without disrupting circadian architecture, motor capacity, or homeostatic hunger, indicating a specific loss of motivational drive. Depleting dFic within subsets of dopamine receptor-expressing (Dop1R1) neurons recapitulates these behavioral alterations. Transcriptomic profiling reveals that dFic depletion does not induce a canonical unfolded protein response, pointing to a specific post-transcriptional alteration rather than global ER stress. At the molecular level, dysregulated dFic alters the localization and abundance of Dop1R1. Finally, we demonstrate that this regulatory axis is evolutionarily conserved, as loss of FICD similarly diminishes motivated behaviors in C. elegans and mice. Together, these findings implicate FICD in the generation of motivated behavior and link dynamic control of ER proteostasis to dopamine-receptor maturation.

neuroscience↗

bioRxiv2026

Label-free identification and classification of leukocyte subpopulations in peripheral blood smears by optical photothermal infrared spectroscopy

The differential count of leukocyte subpopulations from peripheral blood smears remains a cornerstone of clinical hematology but relies on staining protocols or flow cytometry, that introduce variability and obscure native biochemistry. This study presents label-free workflows that distinguish leukocytes from erythrocytes in full blood smears and classifies the leukocyte subtypes as granulocytes, lymphocytes and monocytes using their single cell optical photothermal infrared (OPTIR) spectra. The first approach uses discrete wavenumber images using two key bands at 1546 and 2922 cm-1 to screen leukocytes. The second approach using intrinsic fluorescence imaging of leukocytes offers approximately 1000 times faster screening. Multiple OPTIR spectra are acquired per leukocyte followed by post-acquisition Giemsa staining for cytological cell verification. Full blood smears were prepared on calcium fluoride and standard glass slides to compare substrate effects, and OPTIR spectra were collected with 532 and 785 nm probe lasers to assess probe wavelengths effects. Principal component-linear discriminant analysis separated granulocytes from mononuclear subtypes under all substrate-probe laser combinations tested on the basis of lipid, nucleic-acid and protein bands in OPTIR spectra. Lymphocytes and monocytes were better distinguished on calcium fluoride than on glass. The OPTIR spectra stayed stable over 14 days of storage on both substrates, whereas the intrinsic fluorescence used for cell localization was weak in fresh smears and increased with storage. The non-destructive, fixation-free measurement preserves native protein and lipid composition and allows subsequent cytological staining, supporting OPTIR on routine glass slides for future label-free hematological screening.

biophysics↗

bioRxiv2026

Threshold to brain quantum coherence

Amongst brain structures, we identified a virtual void at the juxta- and paranode, within the intraperiod myelin layers, VIMI , as potential candidate for quantum near-ideal topological insulation. The insulation was extensively weighed here according to current findings. Values of biological and physical boundaries must be exact and a single missing/inexact element defining the anatomy, physiology, biochemistry, cytology, histology, and nano, leads to inaccurate boundaries/quantum equations of the myelin system. We determined the threshold of quantum coherence to decoherence. A propagating internodal electromagnetic interaction through the VIMI void can be represented via Hamiltonian and entanglement. We tested three potential mechanisms for ion K+ data storage. 1. Quantum tunneling through a dielectric, i.e. single myelin layer. 2. Quantum tunneling through squeezing void, VIMI. The action potential is accompanied by mechanical axonal waves and myelin layers compression. Compression reduces the WKB action, and exponentially increases in transmission-tunneling probability. Resonance tunneling may add orders-of-magnitude boosts. 3. Classical squeezing passage. Quantum coherence, tunneling (1) and (2) are theoretically valid and testable, (1) at highly restricted parameter threshold. Unlike quantum tunneling, classical physic passage does not allow fine voltage-controlled metering, sorting, organizing the data storage, later rendering the retrieving impossible.

neuroscience↗

bioRxiv2026

Plant cell-based 4D engineered living materials

Engineered living materials (ELMs) seek to bridge the gap between static engineered matter and dynamic biological systems. Photosynthetic ELMs have emerged as a promising route towards autonomous and sustainable material systems; however, they remain largely based on microorganisms or isolated organelles, whereas the potential of living plant cells as multifunctional, adaptive building blocks remains largely unexplored. Here we establish plant cells as a distinct chassis for engineered living materials and demonstrate that their intrinsic metabolic plasticity, cell-wall biochemistry and mechanics can be harnessed to create adaptive, evolvable and programmable materials. Using Arabidopsis thaliana suspension cells and printable pectin-alginate bioinks, we fabricate two- and three-dimensional living materials that support growth, metabolic plasticity and, long-term viability. Non-photosynthetic ELMs undergo light-induced metabolic reprogramming to acquire photosynthetic activity, demonstrating adaptive behaviour, while continuous cell growth dynamically remodels the mechanical properties of the material, enabling evolution of material properties through intrinsic biological processes. We further exploit endogenous cell wall peroxidases to catalyse the biofabrication of conducting polymers, enabling programmable acquisition of electrochemical functionality without genetic modification. By integrating living plant cells with auxetic and honeycomb architectures, we show that biological activity and structural design jointly determine emergent mechanical behaviour, revealing a principle of geometry-dependent biological reinforcement for living mechanical metamaterials. Centimetre-scale plant ELMs retain robust photosynthetic activity, with electron transport rates approaching those of native Arabidopsis tissues despite the absence of leaf architecture. Together, these findings establish plant cells as versatile building blocks for multifunctional living materials and lay the foundation for self-powered autonomous materials with potential applications in soft robotics, bioelectronics, and carbon capture.

bioengineering↗