Freeze-structuring unlocks minimal-processing strategies for legume texturization

Freeze-structuring unlocks minimal-processing strategies for legume texturization | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Article Freeze-structuring unlocks minimal-processing strategies for legume texturization Andrea Bach*, Elin Perler*, Lenja S. Lemcke, Patrick A. Rühs This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-8663699/v1 This work is licensed under a CC BY 4.0 License Status: Under Review Version 1 posted 13 You are reading this latest preprint version Abstract Legumes are sustainable, nutrient-rich, and affordable, yet their potential remains underused because conventional food processing often fails to utilize the entire raw material or achieve desirable textures. We show that freeze structuring can serve as a minimal-processing technique that imparts texture to legume-based foods without additives or raw material refinement. The method leverages directional ice crystal growth to concentrate starch, protein, and cell wall fragments into aligned interstitial regions as ice forms. Upon thawing, these concentrated domains interlink into a continuous network, reinforcing the gel and producing firmer textures with pronounced anisotropy. This approach was successfully applied to legumes representing 96% of global production, including chickpeas, soybeans, peas, lentils, lupins, common beans, and mung beans, underscoring its versatility. By eliminating fractionation and additive use, freeze structuring offers a decentralized, sustainable solution for producing nutritious, texturized plant-based foods using standard freezing technologies, accessible for industrial and home-scale production worldwide. *Andrea Bach and Elin Perler are co-first authors. Physical sciences/Engineering Physical sciences/Materials science Biological sciences/Plant sciences Freeze structuring legumes plant-based food food structure food texture Figures Figure 1 Figure 2 Figure 3 Figure 4 Main The global food system is at a crucial turning point in adopting sustainable practices to address environmental degradation, resource scarcity, and the growing demand for safe and nutritious food 1 . Plant-based protein alternatives have emerged as a promising solution, offering both environmental and health benefits. Although their popularity increased rapidly over the past decade, recent market slowdowns highlight persistent challenges, including limited consumer acceptance driven by concerns over taste, cost, and excessive processing 2 . Among plant-based protein sources, legumes stand out as a widely recognized opportunity to advance both sustainability and nutrition. They are nutrient-dense, have a low environmental footprint, and serve as key ingredients in many plant-based products. Their ecological adaptability enables cultivation across diverse agro-climatic zones, supported by extensive genetic diversity that facilitates integration into local food systems worldwide 3 , 4 . Legumes provide affordable protein and essential micronutrients such as iron, zinc, and vitamins, making them particularly valuable in low- and middle-income countries where animal protein is scarce, and nutrient deficiencies remain widespread. Beyond addressing global malnutrition, legumes are also gaining relevance in high-income countries amid growing interest in vegetarian diets and alternative proteins. Despite these advantages, legume consumption remains constrained by cultural habits, sensory preferences, and socio-economic barriers 5 . Food processing can play a critical role in addressing these challenges by improving taste, safety, convenience, nutritional quality, and functional performance, thereby expanding the range of legume-based foods 5 , 6 . However, many existing legume-based products rely on highly refined ingredients and intensive processing, contributing to consumer scepticism associated with perceptions of ultra-processed foods as overly engineered or unnatural 2 , 7 . Limited access to advanced processing technologies in some regions further restricts adoption. Together, these factors hinder the broader integration of legumes into diverse diets, highlighting the need for innovative approaches that use minimally refined ingredients while preserving desirable sensory and nutritional properties 5 , 8 . While processing can enable desirable textures and functionality, dominant texturization technologies such as extrusion present notable limitations. High-moisture extrusion typically depends on protein isolates or concentrates, complex supply chains, and substantial capital investment, and is sensitive to raw material variability. Dry extrusion is more tolerant of minimally refined raw materials but often struggles to deliver desirable textures and remains largely inaccessible to consumers 8 , 9 . Fermentation approaches, such as tempeh production, can enhance nutritional quality by reducing antinutrients and improving digestibility, yet pose food safety and scalability challenges in decentralized or low-tech settings 10 . Consequently, there remains a lack of broadly accessible and safe processing strategies that operate on minimally refined ingredients and can promote both the production and consumption of legume-based foods. Freeze structuring is an emerging processing technology with roots in traditional products such as kōri tofu in Japan and modern commercial applications like Quorn. Long recognized for its ability to influence food structure without excessive heat, freeze structuring has recently regained attention for creating fibrous, meat-like textures in plant-based products 11 – 20 . Despite this potential, its broader adoption has been limited by the complexity of food matrices, comprising proteins, polysaccharides, and lipids that interact across multiple length scales and undergo major structural changes during freezing 21 . As a result, freeze structuring for food has predominantly been carried out using purified ingredients, such as protein concentrates and isolates 11 – 17 , or specialized equipment 11 , 15 , 17 . Advancing freeze structuring beyond refined ingredients toward whole-food systems represents a critical opportunity. However, freeze structuring approaches that combine accessibility, scalability, and compatibility with complex food matrices, while preserving nutritional integrity, are still lacking. Here, we show that freeze structuring provides a simple, scalable, and broadly applicable method for imparting texture to whole-legume gels without additives or purification. By leveraging ice crystal growth to redistribute macromolecular components, this method creates anisotropic, fibrous gels with superior mechanical integrity compared to conventional freezing. Its effectiveness across diverse legumes highlights its potential to democratize legume utilization and enable clean-label, customizable foods. Freezing is widely recognized for extending shelf life, reducing food waste, while preserving nutritional quality, which collectively improves the environmental performance of food systems 22 , 23 . Freeze structuring leverages these established benefits by using existing cold-chain infrastructure, supporting its use as a practical and resource-efficient texturing strategy for whole-legume foods. By bridging a critical gap in accessible processing technologies, this approach opens new opportunities for sustainable diets and plant-based food innovation. Results 3.1. Freeze structuring creates anisotropic structure and texture in legume-based systems Freeze structuring of whole-legume gels yields anisotropic, textured food products (Fig. 1 ). First, legumes are hydrated, causing cotyledon cell swelling, initiating water uptake into starch granules. Mixing disrupts cells and disperses cellular fragments throughout the suspension. Thermal treatment leads to starch gelatinization, protein denaturation, and the simultaneous inactivation of antinutrients. The moulded gel is then subjected to directional freezing, typically by placing it on a cooled surface or insulated in an environment below freezing point, thereby applying a unidirectional temperature gradient. During this step, growing ice crystals exclude starch, protein, and cell wall fragments, concentrating them into aligned domains that reinforce the gel network. After thawing, the ice crystals revert to water, leaving a hydrated, lamellar scaffold that retains the anisotropic alignment formed during freezing and provides a self-supporting texture. Texture development in freeze structuring of cooked legume suspensions is driven by two mechanisms: (1) unidirectional freezing, which imparts alignment, and (2) local concentration increases of legume components through freezing, which enhances gel strength. 3.2. Process conditions define structural outcomes in chickpea gels Freezing conditions strongly affect microstructural organization and component distribution within chickpea gels, with unidirectional freezing producing the most pronounced structural alignment. To investigate the cooling and freezing conditions that promote anisotropic structuring in 12.5 wt% chickpea gels, three regimes were examined (Fig. 2 a-c): (a) refrigeration (R), (b) conventional freezing (F), and (c) directional freezing (DF). Gels cooled under refrigeration (R) exhibited a homogeneous, isotropic microstructure without visible alignment (Fig. 2 a). Cyan-stained spherical domains likely represent swollen starch granules in CLSM (confirmation with iodine staining, S1: Figure S1), and red fluorescence likely corresponds to cell wall and protein fragments, dispersed throughout the matrix. Subsequent drying preserved this isotropic structure, indicating that the absence of directional thermal gradients maintained the original network. Conventional freezing (F) produced partial anisotropy, with local alignment but random orientation across the sample (Fig. 2 b). Starch granules (cyan) lost their discrete morphology, appearing diffuse and irregular. This is consistent with structural disruption and granule disintegration caused by freeze-thawing after gelatinization, also observed in maize starch systems 25 . Regions of concentrated material alternated with large pores formed by ice crystal growth. After drying, fibrous structures and voids of varying orientation were evident, reflecting mild inward thermal gradients that arise when outer layers solidify faster than the core during conventional freezing, allowing ice crystals to grow rather than nucleate uniformly throughout the sample. Directional freezing (DF) generated pronounced anisotropy and phase segregation (Fig. 2 c). Material concentrated along lamellae formed by advancing ice crystals, giving rise to a porous, aligned architecture. Starch-rich regions (cyan) appeared diffuse and were aligned along these lamellae, while protein and cell-wall fragments (red) exhibited less orientation, remaining dispersed between aligned starch domains. After drying, the previously aligned ice regions become clearly visible as lamellar structures interconnected by ridges, a consequence of particle exclusion as ice crystals advanced 26 . DF produced the most distinct structural alignment among all cooling regimes and can be achieved using simple setups, such as placing the moulded legume gels in silicone moulds on a metal plate or in a plastic container with expanded polystyrene foam (e.g. Styrofoam) side insulation, all within a standard household freezer (S2: Figure S2). A practical home guide for inducing anisotropy is provided (S3). 3.3. Freezing-induced particle concentration transforms gel properties Since cooling and freeze-structuring affect the distribution of chickpea components, the mechanical gel properties were assessed using oscillatory shear rheology and single- and double-compression tests of gels formed by refrigeration (R), conventional freezing (F), and directional freezing (DF) (Fig. 3 ). DF, unlike R and F, induces a local increase in the concentration of solid components and promotes the formation of lamellar morphologies, thereby increasing viscoelasticity, hardness, and cohesiveness. Chickpea suspensions formed viscoelastic gels with predominantly elastic behaviour through R, F, and DF. The elastic and viscous moduli (G′ and G″) are higher for F and DF than for R, due to a gel network strengthening effect after thawing of ice crystals, yielding elastic moduli of ~ 5×10⁴ Pa for directionally frozen samples versus ~ 10⁴ Pa for refrigerated gels formed using 12.5 wt% chickpea suspensions (Fig. 3 a). A detailed rheological analysis is provided in the supporting information (S4). Elasticity (G′) scaled linearly with concentration (5.0–20.0 wt%) across all conditions, but the rate of increase varied markedly (Fig. 3 b). Refrigerated chickpea gels (R) showed an average linear rise in G’ of 3.1 kPa/wt%, and conventionally frozen gels (F) a rise in G’ of 3.9 kPa/wt%. In contrast, directionally frozen samples (DF) exhibited a ~ 1.5-2x stronger response in elasticity (6.1 kPa/wt%). Above 10.0 wt%, frozen gels were roughly twice as elastic as refrigerated ones, reflecting a higher density of effective crosslinks n ( \(\:\text{G}{\prime\:}\propto\:\text{n}\cdot\:\text{k}\cdot\:\text{T}\) ). Therefore, freezing promotes gel strength by concentrating solids as ice crystals grow. However, this also reduces the water-holding capacity of the system and leads to syneresis upon thawing, particularly at lower initial concentrations and lower network densities. Despite syneresis, DF gels remained anisotropic and self-supporting, retaining mechanically coherent structures relevant for food applications. Concentrations shown in the graphs refer to the initial prepared suspensions, and elasticity values corrected for syneresis confirm the same trends (regression statistics and syneresis, S5 and S6: Table S1 and S2, Figure S3). 15.0 wt% chickpea gels exhibited distinct stress-strain responses under R, F, and DF conditions, reflecting differences in network integrity and anisotropic reinforcement induced by the cooling protocol (Fig. 3 c). In the initial elastic region (up to ~ 10.0 wt% strain), R exhibited the highest slope, followed by DF parallel, DF perpendicular, and then F, indicating that R initially appears stiffer under small deformations. At higher strains, the curves diverge: R reached its maximum stress at ~ 30% strain, then dipped and plateaued at 15.1 kPa, suggesting structural collapse, whereas DF parallel and perpendicular continued rising to 30.4 kPa and 24.4 kPa, respectively, reflecting sustained strain-hardening and network integrity. F gels showed a consistently lower resistance to deformation and reached a maximum stress of ~ 13.5 kPa. Hardness increased nearly linearly with chickpea concentration across all cooling conditions (Fig. 3 d), reflecting denser network formation at higher solids content. R and F gels did not differ significantly, whereas DF gels were consistently harder, with values ~ 1.5-3x higher than R and F. DF samples prepared with 15.0 to 20.0 wt% compressed parallel to the freezing direction were harder by factors of 1.1 to 1.2 than those compressed perpendicular, confirming pronounced anisotropy. This directional dependence arises from the lamellar microstructure formed during directional freezing: parallel compression engages structurally reinforced domains, whereas perpendicular compression disrupts weaker interlamellar regions. Corrected comparisons accounting for syneresis confirm that, at equivalent effective concentrations, DF gels remain harder than R and F gels (statistics and syneresis correction, S7 and S8: Tables S3-S10, Figure S4 and S5). DF gels exhibited the highest cohesiveness, the ability of a gel to withstand a second deformation after the first, indicating a more elastic and structurally resilient texture compared to R and DF gels (Fig. 3 e-f). R, F, and DF chickpea gels at 15.0 wt% indicated structural weakening and permanent deformation upon repeated compression, reflected in a reduced load-bearing capacity during the second deformation (Fig. 3 e). Cohesiveness remained relatively constant across concentrations, with slightly lower values at 12.5 wt%, but DF gels were consistently more cohesive than R and F (Fig. 3 f), demonstrating that DF gels preserve network connectivity more effectively upon reloading. This suggests a more integrated and elastic internal network in DF gels, while R and F lack the highly aligned, anisotropic architecture needed for efficient load redistribution under repeated strain. DF and R gels maintained high recovery values (≈ 82–84%) across most concentrations, while F gels had significantly lower springiness (≈ 68%), indicating that DF gels recover their height after compression similarly to R gels, but with a more robust internal structure (springiness and statistics, S9-S11: Tables S11-24, Figure S6a). Directionally frozen (DF) chickpea gels exhibit mechanical properties that align closely with those of established protein-based foods while introducing distinctive structural features. Chewiness, which reflects the combined effect of hardness, cohesiveness, and springiness during repeated compression, offers a practical measure of overall bite resistance. The combination of greater hardness and cohesiveness in DF gels resulted in markedly higher chewiness (average of 4.4 N) compared to R and F gels (1.0 and 1.1 N, respectively) (chewiness and statistics, S10 and S12: Figure S6b, Tables S25-S31). DF chickpea gels spanned 0.9 to 10.0 N across 12.5 to 20.0 wt% formulations. At 20.0 wt%, DF gels reached 8.7–10.0 N, closely matching mozzarella (~ 9.7 N) and exceeding Brie (~ 5.7 N), while remaining well below firm tofu (~ 20.2 N). At lower concentrations (12.5–15.0 wt%), chewiness values fell between 0.9 and 2.9 N, similar to silken tofu (~ 0.85 N) (references, S13: Table S32). This progression highlights the tunability of DF gels, from soft tofu-like textures at low concentrations to cheese-like chewiness at higher concentrations. Unlike many conventional products, DF gels exhibit a uniquely aligned network, characterized by visually distinct lamellar domains (Fig. 2 ) and hardness anisotropy ratios exceeding 1.1–1.2. In structured foods, such anisotropic organization can enhance structural complexity, a key factor in creating diverse textural experiences and supporting product differentiation 27 . Freeze structuring thus provides a versatile approach for tailoring the texture and mouthfeel of legume-based gels across a wide range of culinary applications. 3.4. Extending freeze-enabled texturization to diverse legume To assess the broader applicability of freeze-induced structuring of minimally processed legumes, we extended the approach beyond chickpeas to a total of 96% of the legumes produced globally 28 (worldwide legume production, S14: Table S33). Gels made from whole red lentils (RL), green peas (GP), black beans (BB), mung bean (MB), black-eyed peas (BP), and soybeans (SB) were subjected to directional freezing and assessed for their structural alignment and textural properties. The resulting gels revealed apparent differences in textural stability and visual anisotropy (Fig. 4 ). All tested legumes formed anisotropically structured gels at 12.5 wt% solids, confirming the broad applicability of freeze-induced alignment (Fig. 4 ). However, mechanical integrity varied among legume species. Red lentil (RL), black lentil (BL), and mung bean (MB) produced firm, stable gels with minimal syneresis, whereas black bean (BB) yielded slightly softer gels. Soybean (SB) gels at 12.5 wt% had low gel stability but still exhibited pronounced lamellar alignment under directional freezing. The textural properties are likely attributable to the elevated lipid and fibre content in soybeans, which can disrupt starch network formation and compromise overall gel integrity (SB composition, S16: Table 34). Increasing the solids concentration to 15.0, 17.5 or 20.0 wt% improved SB texture and retained structural anisotropy, indicating that mechanical properties can be fine-tuned by adjusting concentration. Furthermore, pH increase prior to heat treatment enhanced gel firmness at 12.5 wt%, with higher pH likely promoting protein unfolding, highlighting the role of protein solubility, particle size, and surface hydrophobicity in textural outcomes 29 . Other legumes, including black lentils, yellow lentils, kidney beans, navy beans, and lupins were also successfully anisotropically structured under DF (S15: Figure S7). Gel strength appeared to be influenced by the protein-to-starch (g/g) ratio of legumes (legume composition, S16: Table S34). Samples with relatively low ratios (e.g., red lentil 0.78, green pea 1.27, mung bean 0.61, black-eyed pea 0.70) formed self-supporting gels with higher mechanical integrity, likely due to sufficient starch availability for gelatinization and formation of a continuous load-bearing network. In contrast, samples with relatively high protein-to-starch ratios, such as soybean (3.39), exhibited weaker gel formation, likely due to limited starch content and protein dominance, which can hinder starch gelatinization and disrupt network continuity. Similar effects have been reported in mixed pea protein-maize starch systems, where increasing protein fractions weakened starch-dominated gel networks 30 . In addition to compositional ratios, starch-specific properties, including granule size, swelling capacity, and amylose to amylopectin ratio, may further modulate gel strength and network organisation, contributing to the observed variability among legumes 30 , 31 . These findings confirm that freeze structuring is broadly applicable across legumes and textural outcomes are strongly influenced by intrinsic composition. Key parameters such as solids concentration, particle size, and protein solubility can be strategically adjusted to tailor mechanical properties without compromising structural anisotropy. Further customization can be achieved by formulating mixtures of legumes with varying gel-forming capacities. Alternatively, process adaptations, such as modifying gelation, homogenization, or freezing conditions, offer further opportunities to fine-tune structure and texture. Freeze structuring enables the development of functional, anisotropic textures without reliance on refinement or purification steps, thereby preserving dietary fibre and other nutritionally and functionally valuable components. In contrast, conventional plant-based processing typically relies on fractionation and refinement to achieve functional products. For example, tofu production requires the separation of okara, resulting in substantial macronutrient redistribution 32 : approximately 85% of dietary fibre, 16% of lipids, and 23% of protein from whole soybeans are removed into okara and whey (compositions, S17, Table S35). Similarly, extrusion-based processes commonly involve ingredient fractionation to obtain functional protein or starch fractions 33 . These steps, although integral to those technologies, can result in nutrient losses and generate side streams 8 , 33 . By avoiding fractionation and refinement, freeze structuring retains the full nutritional profile of the raw material and supports more resource-efficient and sustainable food processing. Conclusion Freeze structuring provides a simple and robust strategy to generate anisotropic, fibrous textures in minimally processed whole-legume gels without the need for purification or additives. Directional freezing transforms legume suspensions into mechanically reinforced, anisotropic gels by redistributing macromolecular components and inducing lamellar architectures, as confirmed by microscopic, rheological, and mechanical analyses. Compared with conventional freezing or refrigeration, freeze-structured systems consistently exhibit higher mechanical strength and directional dependence of texture. The successful application across multiple legumes, including chickpeas, lentils, peas, and various beans, highlights the broad versatility of this approach and its capacity to produce diverse textural outcomes from a variety of raw materials. Beyond its functional benefits, freeze structuring addresses key challenges in sustainable food processing. The method relies on simple freezing protocols that are compatible with existing cold-chain infrastructure and does not require specialized or processing steps beyond standard thermal treatment and freezing. At the household scale, the incremental energy demand of freezing is marginal in an already operating freezer, while at small- and medium-enterprise levels the process remains readily implementable using commonly available freezing systems. In contrast to conventional plant-based structuring technologies that rely on ingredient fractionation and refinement, often accompanied by nutrient losses and side-stream generation, freeze structuring operates directly on minimally processed legumes, thereby retaining dietary fibre and other valuable components. While prior research has primarily focused on purified protein or polysaccharide systems, this work demonstrates that directional freezing can effectively structure unrefined, thermally treated legume suspensions into fibrous gels with tunable mechanical properties. Together, these findings position freeze structuring as a globally relevant and scalable approach, adaptable to a wide range of legumes and processing contexts. Future translation toward consumer and culinary applications will benefit from the development of simple, reproducible recipes that integrate soaking, milling, thermal treatment, and freezing using standard kitchen equipment. The structuring mechanism remains effective in the presence of spices and herbs, enabling flavour customization without compromising texture. Providing clear procedural guidance alongside ingredient flexibility will support the adoption of freeze structuring in everyday practice and reinforce its potential contribution to sustainable, clean-label, and minimally processed food systems. Methods Commercial chickpeas ( Cicer arietinum ) were soaked in excess tap water for 24 h at 4°C. After soaking, the pulses were drained and diluted with water to achieve a dry matter content of 20.0% (w/w), then blended into a homogeneous slurry using a Vitamix Ascent A3500i blender (Vitamix, Ohio, USA). This stock was further diluted to working concentrations of 5.0, 7.5, 10.0, 12.5, 15.0, and 17.5% (w/w). Aliquots (50 mL) were transferred into Falcon tubes and heated in a water bath (Caso SV900 Sous Vide Gare, Caso Design, Arnsberg, Germany) at 90°C for 30 min. Samples were inverted every 30 seconds during the first 10 min and gently inverted every 5 min thereafter to ensure uniform thermal treatment. Three post-gelation conditions were directly applied: (1) Refrigeration (R): Samples were stored at 4°C for 25 hours in 50 mL falcon tubes. (2) Conventional Freezing (F): Samples were pre-cooled at 4°C for 1 h, then frozen in a shock freezer at -15°C for 24 h. (3) Directional freezing (DF): Samples were poured into custom insulated silicone moulds (setup, S2: Figure S2a and b), pre-cooled at 4°C for 1 h, and frozen unidirectionally at -15°C for 24 h using an aluminium plate as a cold source at the mould base. After freezing, all samples were thawed at room temperature for 4 h prior to analysis. The same procedure was performed on other legumes, including soybeans ( Glycine max ), green peas ( Pisum sativum) , black-eyed pea ( Vigna unguiculata ), red, black and yellow lentils ( Lens culinaris ), mung beans ( Vigna radiata ), lupins ( Lupinus albus ), and common beans ( Phaseolus vulgaris ) namely black beans, kidney beans, navy beans. 5.1. Rheology Amplitude sweeps were performed on chickpea gels prepared under the three structuring conditions (R, F, DF) using a rheometer (MCR 302, Anton Paar GmbH, Graz, Austria) with a plate-plate geometry (PP25), with both upper and lower plates roughened, and a gap of 1 mm at 25°C. Strain was increased from 0.01% to 300% at a constant frequency of 1 rad s⁻¹. From these sweeps, the plateau storage modulus (G′) and the limit of the linear viscoelastic region (LVER) was determined as the strain at which the storage modulus G' first dropped below 95% of its initial plateau value. 5.2. Texture Analysis Texture was evaluated using single- and double-compression tests on a texture analyzer (TA.XTplus, Stable Micro Systems, Surrey, UK) equipped with the Exponent software (v6.2.6.0). Samples were equilibrated to room temperature and cut into cubes of 15 mm edge length using a sharp kitchen knife. R, F, and DF chickpea gels at 12.5, 15.0, 17.5, and 20.0 wt% were tested. Texture Profile Analysis (TPA) was performed with a 40 mm flat probe by compressing each sample twice with a 5 kg load cell, with DF measured parallel and perpendicular to the temperature gradient. Pre-test, test, and post-test speeds of 300, 180, and 300 mm min⁻¹ were applied, respectively. Each compression reached 50% of the original sample height, with a 5 s interval between compressions. All measurements were performed in triplicate, using cubes taken from the middle section of each sample’s height. For reference values, the same procedure was used to test commercial Brie cheese, mozzarella, silken tofu, and firm tofu. Hardness was defined as the maximum force during the first compression, springiness as the ratio of sample height recovered between the first and second compression, cohesiveness as the ratio of the positive area of the second compression to that of the first compression, and chewiness as hardness × cohesiveness × springiness. 5.3. Confocal Laser Scanning Microscopy (CLSM) R, F, and DF gels were cut into thin slices of approximately 1 mm using a razor blade. The CLSM method was adapted from Lyu et al. (2022) 30 . A staining solution containing fluorescein isothiocyanate isomer I (FITC, CAS 3326-32-7, Sigma-Aldrich, St. Louis, USA) and rhodamine B (CAS 81-88-9, Sigma-Aldrich, St. Louis, USA) (each at 0.0025 w/v%) in N,N-dimethylformamide (DMF, anhydrous, 99.8%, CAS 68-12-2, Sigma-Aldrich, Germany) was used. Gel slices were incubated in the dye solution for 1 hour, followed by three washes with deionized water over a total of 30 minutes to remove excess dye. Confocal imaging was performed on a Leica TCS SP8 (Leica Microsystems, Wetzlar, Germany) using a 10× objective (0.32 NA, HC PL FLUOTAR). Z-stacks were acquired for each sample (100 slices, 1 µm spacing) over a representative region. FITC was excited with a 488 nm laser and detected between 500–530 nm. Rhodamine B was excited with a 552 nm laser and detected between 584–620 nm. Images were adjusted for brightness and contrast. 5.4. Photographic images Photographic images were taking using a Canon EOS 70D (Canon Inc., Tokyo, Japan) mounted inside a photobox with constant lighting conditions. Images were adjusted for brightness and contrast. 5.5. Digital Microscopy (DM) F, and DF gels were cut along the freezing temperature gradient into 3–4 mm slices using a carpet knife and freeze-dried for 24 h in a freeze dryer (FreeZone 4.5 L, Labconco, Kansas City, USA) connected to a rotary vane vacuum pump (117 L min⁻¹, 230 V, Labconco, Kansas City, USA) at -84°C. For refrigerated gels, a slice of the same dimensions was immersed in liquid nitrogen and subsequently freeze-dried. Samples were imaged using a digital microscope (VHX-X1F, Keyence, Osaka, Japan) with a VH-Z20T lens and side-light adapter. Images were acquired at 100× and 150× magnification, stitched, and adjusted for brightness and contrast. 5.6. Scanning Electron Microscopy (SEM) DF and F chickpea gels were cut across the freezing gradient into 3–4 mm slices using a carpet knife and freeze-dried for 24 h in a freeze dryer (FreeZone 4.5 L, Labconco, Kansas City, USA) connected to a rotary vane vacuum pump (117 L min⁻¹, 230 V, Labconco, Kansas City, USA) at -84°C. For refrigerated gels, a slice of the same dimensions was immersed in liquid nitrogen and subsequently freeze-dried. Freeze-dried samples were trimmed into 4–5 mm cubes and mounted on SEM stubs with the cut surface (perpendicular to the temperature gradient) facing upward. A 4 nm platinum coating was applied using a compact coating unit (CCU-010, Safematic GmbH, Switzerland). Imaging was performed on a GeminiSEM 450 (Carl Zeiss Microscopy GmbH, Jena, Germany) equipped with an SE2 detector, using an acceleration voltage of 2.00 kV and an aperture current of 100 pA. Declarations Acknowledgements The chair of Food Structure Engineering gratefully acknowledges financial support from Nestlé, Bühler, and Givaudan via the ETH Zürich Foundation. The funders played no role in study design, data collection, analysis and interpretation of data, or the writing of this manuscript. The authors thank Marco Burkolter (Food Microbiology Group, ETH Zürich) for help with CLSM and Mathias Steinacher (Complex Materials Group, ETH Zürich) for his assistance in CLSM and SEM. Author information Affiliation Department of Health Sciences and Technology, Institute of Food, Nutrition and Health, ETH Zürich, Schmelzbergstrasse 7, Zürich, 8092, Zürich, Switzerland. Contribution A.B., L.L., E.P., and P.R. conceptualized the work. A.B., E.P., L.L., and P.R. designed the methodology. A.B., E.P., and L.L. performed data collection. A.B., E.P., and P.R. wrote the original draft. A.B. created the figures. A.B., E.P., L.L. and P.R. revised the final version of the paper. Corresponding author(s) Patrick A. Rühs | [email protected] Competing interests The authors declare no competing interests. References Rockström, J. et al. The EAT–Lancet Commission on healthy, sustainable, and just food systems. The Lancet 406, 1625–1700 (2025). Caputo, V., Sun, J., Staples, A. J. & Taylor, H. Market outlook for meat alternatives: challenges, opportunities, and new developments. Trends Food Sci Technol 148, 104474 (2024). FAO/OCCP. Pulses: Nutritious Seeds for a Sustainable Future . (FAO, 2016). doi:10.4060/I5528E. Vaz Patto, M. C. et al. Achievements and challenges in improving the nutritional quality of food legumes. CRC Crit Rev Plant Sci 34, 105–143 (2015). Lisciani, S. et al. Legumes and common beans in sustainable diets: nutritional quality, environmental benefits, spread and use in food preparations. Front Nutr 11, 1385232 (2024). Hammer, L. et al. Influence of processing on protein quality and environmental impact assessment of soy-based meat analogues. Food Research International 222, 117636 (2025). Forde, C. G. Beyond ultra-processed: considering the future role of food processing in human health. Proceedings of the Nutrition Society 82, 406–418 (2023). Germerdonk, T., Bach, A., Marangoni, A. G., Mishra, K. & Rühs, P. A. Unrefined plant raw materials are key to nutritious food. Nat Food 6, 657–663 (2025). Del Rio, A. R., Boom, R. M. & Janssen, A. E. M. Effect of fractionation and processing conditions on the digestibility of plant proteins as food ingredients. Foods 2022, Vol. 11, Page 870 11, 870 (2022). Yee, C. S. et al. Smart fermentation technologies: microbial process control in traditional fermented foods. Fermentation 2025, Vol. 11, Page 323 11, 323 (2025). De Henau, R., de Vries, A. & Rousseau, D. Structure and mechanical properties of anisotropic agar gels obtained via unidirectional freezing. Food Research International 114626 (2024). Rühs, P., Müller, M. & Savorani, L. A. Freeze structured and enzymatically crosslinked food materials. (2024). US Patent 2024/0415147 A1 Bryson, C., Rousseau, D., De Vries, A. & Gregson, C. M. Process for producing cookable, fibrous meat analogues with directional freezing. (2022). US Patent 11,241,024 B1 Basse, B., El Chemali, M. L., Dupuis, H. & Masbernat, L. Fibrous or laminated, and textured food product and method for producing same. (2024). Fischer, J., Bender, D., Domig, K. J. & Fuhrmann, P. L. Inducing anisotropy in emulsion-filled hydrogels by unidirectional freezing. Food Hydrocoll 162, 111008 (2025). Nakagawa, K., Chantanuson, R., Boonarsa, P., Seephua, N. & Siriamornpun, S. Meat analogue preparation from cricket and rice powder mixtures with controlled textural and nutritional quality by freeze alignment technique. Food Chem X 22, 101402 (2024). Ryu, J. & McClements, D. J. Freeze/thaw-triggered fixation of directionally frozen plant-based food matrices: Controlled release of gelling agents using double emulsions. Food Hydrocoll 173, 112308 (2026). Sengar, A. S. et al. Developing freeze-structured meat alternatives using pea and faba proteins: Evaluating their partial and complete substitution in beef patties. Food Structure 45, 100451 (2025). Durage, T. T. D. et al. Developing a chickpea protein–flaxseed oil emulsion gel meat analogue using the freeze-alignment technique. Sustainable Food Technology 3, 1996–2008 (2025). Chantanuson, R., Nagamine, S., Kobayashi, T. & Nakagawa, K. Effect of dry heat treatment of soy protein powder on aligned structure formation in soy protein-based food gels during freezing. J Food Eng 363, 111779 (2024). Chantanuson, R., Nagamine, S., Kobayashi, T. & Nakagawa, K. Preparation of soy protein-based food gels and control of fibrous structure and rheological property by freezing. Food Structure 32, 100258 (2022). Xu, L., Li, R. & Roe, B. Frozen food purchasing and home freezing of fresh foods: associations with household food waste. British Food Journal 126, 4260–4276 (2024). Barbosa-Cánovas, G. V., Altunakar, B. & Mejia-Lorio, D. J. Freezing of Fruits and Vegetables: An Agri-Business Alternative for Rural and Semi-Rural Areas (2005). Xiong, W., Devkota, L., Zhang, B., Muir, J. & Dhital, S. Intact cells: “Nutritional capsules” in plant foods. Compr Rev Food Sci Food Saf 21, 1198–1217 (2022). Zhang, C. et al. Critical melting assisted freeze-thawing treatment as a novel clean-label way to prepare porous starch: Synergistic effect of melting and ice recrystallization. Food Hydrocoll 131, 107730 (2022). Bach, A. & Rühs, P. A. Addressing multifactorial complexity in freeze structuring of food colloids. Curr Opin Colloid Interface Sci 101941 (2025). Oppen, D., Grossmann, L. & Weiss, J. Insights into characterizing and producing anisotropic food structures. Crit Rev Food Sci Nutr 64, 1158–1176 (2024). FAO. FAOSTAT: Crops and livestock products. https://www.fao.org/faostat/en/#data/QCL. Licence: CC-BY-4.0 (2023). Sun, P. et al. Improving gel properties of soy protein isolate through alkaline pH-shifting, mild heat treatment, and TGase cross-linking. Food Hydrocoll 144, 108924 (2023). Lyu, Z., Sala, G. & Scholten, E. Water distribution in maize starch-pea protein gels as determined by a novel confocal laser scanning microscopy image analysis method and its effect on structural and mechanical properties of composite gels. Food Hydrocoll 133, 107942 (2022). Cornejo-Ramírez, Y. I. et al. The structural characteristics of starches and their functional properties. CYTA - Journal of Food 16, 1003–1017 (2018). van der Riet, W. B., Wight, A. W., Cilliers, J. J. L. & Datel, J. M. Food chemical investigation of tofu and its byproduct okara. Food Chem 34, 193–202 (1989). Del Rio, A. R., Boom, R. M. & Janssen, A. E. M. Effect of fractionation and processing conditions on the digestibility of plant proteins as food ingredients. Foods 2022, Vol. 11, Page 870 11, 870 (2022). Additional Declarations No competing interests reported. Supplementary Files FSLegumesSupporting.docx Cite Share Download PDF Status: Under Review Version 1 posted Editorial decision: Revision requested 28 Feb, 2026 Reviews received at journal 27 Feb, 2026 Reviews received at journal 26 Feb, 2026 Reviews received at journal 25 Feb, 2026 Reviewers agreed at journal 08 Feb, 2026 Reviews received at journal 08 Feb, 2026 Reviewers agreed at journal 08 Feb, 2026 Reviewers agreed at journal 07 Feb, 2026 Reviewers agreed at journal 07 Feb, 2026 Reviewers invited by journal 07 Feb, 2026 Editor assigned by journal 25 Jan, 2026 Submission checks completed at journal 25 Jan, 2026 First submitted to journal 21 Jan, 2026 You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-8663699","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":588937121,"identity":"fef7f2d6-e48d-4c58-b2ec-639175e1adfb","order_by":0,"name":"Andrea Bach*","email":"","orcid":"","institution":"Institute of Food, Nutrition, and Health, Department of Health Sciences and Technology, ETH Zürich, Switzerland","correspondingAuthor":false,"prefix":"","firstName":"Andrea","middleName":"","lastName":"Bach*","suffix":""},{"id":588937122,"identity":"d905e618-24cb-4ccf-b992-21d58ff7bd96","order_by":1,"name":"Elin Perler*","email":"","orcid":"","institution":"Institute of Food, Nutrition, and Health, Department of Health Sciences and Technology, ETH Zürich, Switzerland","correspondingAuthor":false,"prefix":"","firstName":"Elin","middleName":"","lastName":"Perler*","suffix":""},{"id":588937123,"identity":"724a0cd4-67c5-40c7-8770-486ec36fcb66","order_by":2,"name":"Lenja S. Lemcke","email":"","orcid":"","institution":"Institute of Food, Nutrition, and Health, Department of Health Sciences and Technology, ETH Zürich, Switzerland","correspondingAuthor":false,"prefix":"","firstName":"Lenja","middleName":"S.","lastName":"Lemcke","suffix":""},{"id":588937120,"identity":"df141c91-a223-48a5-9f85-cbb0902dc56b","order_by":3,"name":"Patrick A. Rühs","email":"data:image/png;base64,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","orcid":"","institution":"Institute of Food, Nutrition, and Health, Department of Health Sciences and Technology, ETH Zürich, Switzerland","correspondingAuthor":true,"prefix":"","firstName":"Patrick","middleName":"A.","lastName":"Rühs","suffix":""}],"badges":[],"createdAt":"2026-01-21 22:23:45","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-8663699/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-8663699/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":102602399,"identity":"84410511-0fe0-42c1-a26d-ce5c1bcd9149","added_by":"auto","created_at":"2026-02-13 13:18:46","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":3027105,"visible":true,"origin":"","legend":"\u003cp\u003eFreeze structuring process for legume-based food products. Freeze structuring of whole-legume gels creates anisotropic, textured products. Legume hydration swells cells and starch granules, mixing disrupts cells and disperses cellular fragments, heating causes starch gelatinization and protein denaturation. Directional freezing concentrates legume components into aligned domains. After thawing, a lamellar scaffold with self-supporting texture results. Figure inspired by Xiong et al. (2022)\u003csup\u003e24\u003c/sup\u003e and created with BioRender.\u003c/p\u003e","description":"","filename":"floatimage1.png","url":"https://assets-eu.researchsquare.com/files/rs-8663699/v1/56c3ef2fbff6155b4becd0ba.png"},{"id":102602396,"identity":"d4b0a85a-c7bd-40e5-9e25-afcf2edd2ae7","added_by":"auto","created_at":"2026-02-13 13:18:45","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":12399827,"visible":true,"origin":"","legend":"\u003cp\u003eEffect of cooling process on component distribution and microstructure in chickpea-based gels. Photographic images of chickpea gel samples, confocal laser scanning micrographs (CLSM) of chickpea gels stained with FITC (cyan) and rhodamine B (red), digital micrographs (DM), and scanning electron micrographs (SEM) of (a) refrigerated (R), (b) conventionally frozen (F), and (c) directionally frozen (DF) chickpea gels at 12.5 wt%.\u003c/p\u003e","description":"","filename":"floatimage2.png","url":"https://assets-eu.researchsquare.com/files/rs-8663699/v1/4c870437484a502abc6edcda.png"},{"id":102602394,"identity":"39e4726d-fdf7-49ed-9487-6d24b84b0ac8","added_by":"auto","created_at":"2026-02-13 13:18:45","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":4972965,"visible":true,"origin":"","legend":"\u003cp\u003eRheological and mechanical properties of chickpea gels subjected to different cooling and freezing regimes. (a) Oscillatory amplitude sweeps of 12.5 wt% chickpea (CP) gels: refrigerated (R), conventionally frozen (F), and directionally frozen (DF). (b) G′ in the linear viscoelastic region (LVER) of amplitude sweeps of R, F, and DF gels from 5.0 to 20.0 wt%. (c) Stress-strain curves obtained from uniaxial compression of gels at 15.0 wt%, with DF measured parallel and perpendicular to the temperature gradient. (d) Hardness of R, F, and DF gels with concentrations from 12.5 to 20.0 wt%. (e) Stress-strain curves obtained from double compression of gels at 15.0 wt%, with DF measured parallel and perpendicular to the temperature gradient. (f) Cohesiveness of R, F, and DF gels with concentrations from 12.5 to 20.0 wt%.\u003c/p\u003e","description":"","filename":"floatimage3.png","url":"https://assets-eu.researchsquare.com/files/rs-8663699/v1/27c0db4ee55135f15a8d744e.png"},{"id":102747225,"identity":"38a237aa-5582-4d96-9b3b-912c3da4d12c","added_by":"auto","created_at":"2026-02-16 09:04:13","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":4472956,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eFreeze-structured legumes.\u003c/strong\u003e Photographic images of gels prepared from whole red lentils (RL), green peas (GP), black beans (BB), mung beans (MB), black-eyed pea (BP) and soybeans (SB) at 12.5 wt% solids after thermal treatment and directional freezing. Additional samples include soybean (SB) gels at elevated concentrations (15.0 to 20.0 wt%) and pH-adjusted SB gels. Corresponding digital micrographs were obtained after freeze-drying to visualize structural alignment.\u003c/p\u003e","description":"","filename":"floatimage4.png","url":"https://assets-eu.researchsquare.com/files/rs-8663699/v1/1f37c652ea2e183f6bd677bb.png"},{"id":105562637,"identity":"e224896e-5b9f-441d-8b67-29933eaeb434","added_by":"auto","created_at":"2026-03-27 12:43:54","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":25497352,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-8663699/v1/1c7e3833-13cb-49a8-8173-de707cc4b0fe.pdf"},{"id":102602398,"identity":"08d74167-cb34-411a-82ac-0bf1e57f4935","added_by":"auto","created_at":"2026-02-13 13:18:46","extension":"docx","order_by":0,"title":"","display":"","copyAsset":false,"role":"supplement","size":16542630,"visible":true,"origin":"","legend":"","description":"","filename":"FSLegumesSupporting.docx","url":"https://assets-eu.researchsquare.com/files/rs-8663699/v1/1a2fc217374a1b55b7cf2d47.docx"}],"financialInterests":"No competing interests reported.","formattedTitle":"Freeze-structuring unlocks minimal-processing strategies for legume texturization","fulltext":[{"header":"Main","content":"\u003cp\u003eThe global food system is at a crucial turning point in adopting sustainable practices to address environmental degradation, resource scarcity, and the growing demand for safe and nutritious food\u003csup\u003e\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e\u003c/sup\u003e. Plant-based protein alternatives have emerged as a promising solution, offering both environmental and health benefits. Although their popularity increased rapidly over the past decade, recent market slowdowns highlight persistent challenges, including limited consumer acceptance driven by concerns over taste, cost, and excessive processing\u003csup\u003e\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e\u003c/sup\u003e.\u003c/p\u003e \u003cp\u003eAmong plant-based protein sources, legumes stand out as a widely recognized opportunity to advance both sustainability and nutrition. They are nutrient-dense, have a low environmental footprint, and serve as key ingredients in many plant-based products. Their ecological adaptability enables cultivation across diverse agro-climatic zones, supported by extensive genetic diversity that facilitates integration into local food systems worldwide\u003csup\u003e\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e,\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e\u003c/sup\u003e. Legumes provide affordable protein and essential micronutrients such as iron, zinc, and vitamins, making them particularly valuable in low- and middle-income countries where animal protein is scarce, and nutrient deficiencies remain widespread. Beyond addressing global malnutrition, legumes are also gaining relevance in high-income countries amid growing interest in vegetarian diets and alternative proteins. Despite these advantages, legume consumption remains constrained by cultural habits, sensory preferences, and socio-economic barriers\u003csup\u003e\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e\u003c/sup\u003e.\u003c/p\u003e \u003cp\u003eFood processing can play a critical role in addressing these challenges by improving taste, safety, convenience, nutritional quality, and functional performance, thereby expanding the range of legume-based foods\u003csup\u003e\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e,\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e\u003c/sup\u003e. However, many existing legume-based products rely on highly refined ingredients and intensive processing, contributing to consumer scepticism associated with perceptions of ultra-processed foods as overly engineered or unnatural\u003csup\u003e\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e,\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e\u003c/sup\u003e. Limited access to advanced processing technologies in some regions further restricts adoption. Together, these factors hinder the broader integration of legumes into diverse diets, highlighting the need for innovative approaches that use minimally refined ingredients while preserving desirable sensory and nutritional properties\u003csup\u003e\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e,\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e\u003c/sup\u003e.\u003c/p\u003e \u003cp\u003eWhile processing can enable desirable textures and functionality, dominant texturization technologies such as extrusion present notable limitations. High-moisture extrusion typically depends on protein isolates or concentrates, complex supply chains, and substantial capital investment, and is sensitive to raw material variability. Dry extrusion is more tolerant of minimally refined raw materials but often struggles to deliver desirable textures and remains largely inaccessible to consumers\u003csup\u003e\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e,\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e\u003c/sup\u003e. Fermentation approaches, such as tempeh production, can enhance nutritional quality by reducing antinutrients and improving digestibility, yet pose food safety and scalability challenges in decentralized or low-tech settings\u003csup\u003e\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e\u003c/sup\u003e. Consequently, there remains a lack of broadly accessible and safe processing strategies that operate on minimally refined ingredients and can promote both the production and consumption of legume-based foods.\u003c/p\u003e \u003cp\u003eFreeze structuring is an emerging processing technology with roots in traditional products such as kōri tofu in Japan and modern commercial applications like Quorn. Long recognized for its ability to influence food structure without excessive heat, freeze structuring has recently regained attention for creating fibrous, meat-like textures in plant-based products\u003csup\u003e\u003cspan additionalcitationids=\"CR12 CR13 CR14 CR15 CR16 CR17 CR18 CR19\" citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e\u003c/sup\u003e. Despite this potential, its broader adoption has been limited by the complexity of food matrices, comprising proteins, polysaccharides, and lipids that interact across multiple length scales and undergo major structural changes during freezing\u003csup\u003e\u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e21\u003c/span\u003e\u003c/sup\u003e. As a result, freeze structuring for food has predominantly been carried out using purified ingredients, such as protein concentrates and isolates\u003csup\u003e\u003cspan additionalcitationids=\"CR12 CR13 CR14 CR15 CR16\" citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e\u003c/sup\u003e, or specialized equipment\u003csup\u003e\u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e,\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e,\u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e\u003c/sup\u003e. Advancing freeze structuring beyond refined ingredients toward whole-food systems represents a critical opportunity. However, freeze structuring approaches that combine accessibility, scalability, and compatibility with complex food matrices, while preserving nutritional integrity, are still lacking.\u003c/p\u003e \u003cp\u003eHere, we show that freeze structuring provides a simple, scalable, and broadly applicable method for imparting texture to whole-legume gels without additives or purification. By leveraging ice crystal growth to redistribute macromolecular components, this method creates anisotropic, fibrous gels with superior mechanical integrity compared to conventional freezing. Its effectiveness across diverse legumes highlights its potential to democratize legume utilization and enable clean-label, customizable foods. Freezing is widely recognized for extending shelf life, reducing food waste, while preserving nutritional quality, which collectively improves the environmental performance of food systems\u003csup\u003e\u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e22\u003c/span\u003e,\u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e23\u003c/span\u003e\u003c/sup\u003e. Freeze structuring leverages these established benefits by using existing cold-chain infrastructure, supporting its use as a practical and resource-efficient texturing strategy for whole-legume foods. By bridging a critical gap in accessible processing technologies, this approach opens new opportunities for sustainable diets and plant-based food innovation.\u003c/p\u003e"},{"header":"Results","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003e3.1. Freeze structuring creates anisotropic structure and texture in legume-based systems\u003c/h2\u003e \u003cp\u003eFreeze structuring of whole-legume gels yields anisotropic, textured food products (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). First, legumes are hydrated, causing cotyledon cell swelling, initiating water uptake into starch granules. Mixing disrupts cells and disperses cellular fragments throughout the suspension. Thermal treatment leads to starch gelatinization, protein denaturation, and the simultaneous inactivation of antinutrients. The moulded gel is then subjected to directional freezing, typically by placing it on a cooled surface or insulated in an environment below freezing point, thereby applying a unidirectional temperature gradient. During this step, growing ice crystals exclude starch, protein, and cell wall fragments, concentrating them into aligned domains that reinforce the gel network. After thawing, the ice crystals revert to water, leaving a hydrated, lamellar scaffold that retains the anisotropic alignment formed during freezing and provides a self-supporting texture. Texture development in freeze structuring of cooked legume suspensions is driven by two mechanisms: (1) unidirectional freezing, which imparts alignment, and (2) local concentration increases of legume components through freezing, which enhances gel strength.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec4\" class=\"Section2\"\u003e \u003ch2\u003e3.2. Process conditions define structural outcomes in chickpea gels\u003c/h2\u003e \u003cp\u003eFreezing conditions strongly affect microstructural organization and component distribution within chickpea gels, with unidirectional freezing producing the most pronounced structural alignment. To investigate the cooling and freezing conditions that promote anisotropic structuring in 12.5 wt% chickpea gels, three regimes were examined (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003ea-c): (a) refrigeration (R), (b) conventional freezing (F), and (c) directional freezing (DF).\u003c/p\u003e \u003cp\u003eGels cooled under refrigeration (R) exhibited a homogeneous, isotropic microstructure without visible alignment (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003ea). Cyan-stained spherical domains likely represent swollen starch granules in CLSM (confirmation with iodine staining, S1: Figure S1), and red fluorescence likely corresponds to cell wall and protein fragments, dispersed throughout the matrix. Subsequent drying preserved this isotropic structure, indicating that the absence of directional thermal gradients maintained the original network.\u003c/p\u003e \u003cp\u003eConventional freezing (F) produced partial anisotropy, with local alignment but random orientation across the sample (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003eb). Starch granules (cyan) lost their discrete morphology, appearing diffuse and irregular. This is consistent with structural disruption and granule disintegration caused by freeze-thawing after gelatinization, also observed in maize starch systems\u003csup\u003e\u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e25\u003c/span\u003e\u003c/sup\u003e. Regions of concentrated material alternated with large pores formed by ice crystal growth. After drying, fibrous structures and voids of varying orientation were evident, reflecting mild inward thermal gradients that arise when outer layers solidify faster than the core during conventional freezing, allowing ice crystals to grow rather than nucleate uniformly throughout the sample.\u003c/p\u003e \u003cp\u003eDirectional freezing (DF) generated pronounced anisotropy and phase segregation (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003ec). Material concentrated along lamellae formed by advancing ice crystals, giving rise to a porous, aligned architecture. Starch-rich regions (cyan) appeared diffuse and were aligned along these lamellae, while protein and cell-wall fragments (red) exhibited less orientation, remaining dispersed between aligned starch domains. After drying, the previously aligned ice regions become clearly visible as lamellar structures interconnected by ridges, a consequence of particle exclusion as ice crystals advanced\u003csup\u003e\u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e26\u003c/span\u003e\u003c/sup\u003e. DF produced the most distinct structural alignment among all cooling regimes and can be achieved using simple setups, such as placing the moulded legume gels in silicone moulds on a metal plate or in a plastic container with expanded polystyrene foam (e.g. Styrofoam) side insulation, all within a standard household freezer (S2: Figure S2). A practical home guide for inducing anisotropy is provided (S3).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec5\" class=\"Section2\"\u003e \u003ch2\u003e3.3. Freezing-induced particle concentration transforms gel properties\u003c/h2\u003e \u003cp\u003eSince cooling and freeze-structuring affect the distribution of chickpea components, the mechanical gel properties were assessed using oscillatory shear rheology and single- and double-compression tests of gels formed by refrigeration (R), conventional freezing (F), and directional freezing (DF) (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003e). DF, unlike R and F, induces a local increase in the concentration of solid components and promotes the formation of lamellar morphologies, thereby increasing viscoelasticity, hardness, and cohesiveness.\u003c/p\u003e \u003cp\u003eChickpea suspensions formed viscoelastic gels with predominantly elastic behaviour through R, F, and DF. The elastic and viscous moduli (G\u0026prime; and G\u0026Prime;) are higher for F and DF than for R, due to a gel network strengthening effect after thawing of ice crystals, yielding elastic moduli of ~\u0026thinsp;5\u0026times;10⁴ Pa for directionally frozen samples versus ~\u0026thinsp;10⁴ Pa for refrigerated gels formed using 12.5 wt% chickpea suspensions (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003ea). A detailed rheological analysis is provided in the supporting information (S4).\u003c/p\u003e \u003cp\u003eElasticity (G\u0026prime;) scaled linearly with concentration (5.0\u0026ndash;20.0 wt%) across all conditions, but the rate of increase varied markedly (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003eb). Refrigerated chickpea gels (R) showed an average linear rise in G\u0026rsquo; of 3.1 kPa/wt%, and conventionally frozen gels (F) a rise in G\u0026rsquo; of 3.9 kPa/wt%. In contrast, directionally frozen samples (DF) exhibited a\u0026thinsp;~\u0026thinsp;1.5-2x stronger response in elasticity (6.1 kPa/wt%). Above 10.0 wt%, frozen gels were roughly twice as elastic as refrigerated ones, reflecting a higher density of effective crosslinks n (\u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(\\:\\text{G}{\\prime\\:}\\propto\\:\\text{n}\\cdot\\:\\text{k}\\cdot\\:\\text{T}\\)\u003c/span\u003e\u003c/span\u003e). Therefore, freezing promotes gel strength by concentrating solids as ice crystals grow. However, this also reduces the water-holding capacity of the system and leads to syneresis upon thawing, particularly at lower initial concentrations and lower network densities. Despite syneresis, DF gels remained anisotropic and self-supporting, retaining mechanically coherent structures relevant for food applications. Concentrations shown in the graphs refer to the initial prepared suspensions, and elasticity values corrected for syneresis confirm the same trends (regression statistics and syneresis, S5 and S6: Table S1 and S2, Figure S3).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003e15.0 wt% chickpea gels exhibited distinct stress-strain responses under R, F, and DF conditions, reflecting differences in network integrity and anisotropic reinforcement induced by the cooling protocol (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003ec). In the initial elastic region (up to ~\u0026thinsp;10.0 wt% strain), R exhibited the highest slope, followed by DF parallel, DF perpendicular, and then F, indicating that R initially appears stiffer under small deformations. At higher strains, the curves diverge: R reached its maximum stress at ~\u0026thinsp;30% strain, then dipped and plateaued at 15.1 kPa, suggesting structural collapse, whereas DF parallel and perpendicular continued rising to 30.4 kPa and 24.4 kPa, respectively, reflecting sustained strain-hardening and network integrity. F gels showed a consistently lower resistance to deformation and reached a maximum stress of ~\u0026thinsp;13.5 kPa.\u003c/p\u003e \u003cp\u003eHardness increased nearly linearly with chickpea concentration across all cooling conditions (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003ed), reflecting denser network formation at higher solids content. R and F gels did not differ significantly, whereas DF gels were consistently harder, with values\u0026thinsp;~\u0026thinsp;1.5-3x higher than R and F. DF samples prepared with 15.0 to 20.0 wt% compressed parallel to the freezing direction were harder by factors of 1.1 to 1.2 than those compressed perpendicular, confirming pronounced anisotropy. This directional dependence arises from the lamellar microstructure formed during directional freezing: parallel compression engages structurally reinforced domains, whereas perpendicular compression disrupts weaker interlamellar regions. Corrected comparisons accounting for syneresis confirm that, at equivalent effective concentrations, DF gels remain harder than R and F gels (statistics and syneresis correction, S7 and S8: Tables S3-S10, Figure S4 and S5).\u003c/p\u003e \u003cp\u003eDF gels exhibited the highest cohesiveness, the ability of a gel to withstand a second deformation after the first, indicating a more elastic and structurally resilient texture compared to R and DF gels (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003ee-f). R, F, and DF chickpea gels at 15.0 wt% indicated structural weakening and permanent deformation upon repeated compression, reflected in a reduced load-bearing capacity during the second deformation (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003ee). Cohesiveness remained relatively constant across concentrations, with slightly lower values at 12.5 wt%, but DF gels were consistently more cohesive than R and F (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003ef), demonstrating that DF gels preserve network connectivity more effectively upon reloading. This suggests a more integrated and elastic internal network in DF gels, while R and F lack the highly aligned, anisotropic architecture needed for efficient load redistribution under repeated strain. DF and R gels maintained high recovery values (\u0026asymp;\u0026thinsp;82\u0026ndash;84%) across most concentrations, while F gels had significantly lower springiness (\u0026asymp;\u0026thinsp;68%), indicating that DF gels recover their height after compression similarly to R gels, but with a more robust internal structure (springiness and statistics, S9-S11: Tables S11-24, Figure S6a).\u003c/p\u003e \u003cp\u003eDirectionally frozen (DF) chickpea gels exhibit mechanical properties that align closely with those of established protein-based foods while introducing distinctive structural features. Chewiness, which reflects the combined effect of hardness, cohesiveness, and springiness during repeated compression, offers a practical measure of overall bite resistance. The combination of greater hardness and cohesiveness in DF gels resulted in markedly higher chewiness (average of 4.4 N) compared to R and F gels (1.0 and 1.1 N, respectively) (chewiness and statistics, S10 and S12: Figure S6b, Tables S25-S31). DF chickpea gels spanned 0.9 to 10.0 N across 12.5 to 20.0 wt% formulations. At 20.0 wt%, DF gels reached 8.7\u0026ndash;10.0 N, closely matching mozzarella (~\u0026thinsp;9.7 N) and exceeding Brie (~\u0026thinsp;5.7 N), while remaining well below firm tofu (~\u0026thinsp;20.2 N). At lower concentrations (12.5\u0026ndash;15.0 wt%), chewiness values fell between 0.9 and 2.9 N, similar to silken tofu (~\u0026thinsp;0.85 N) (references, S13: Table S32). This progression highlights the tunability of DF gels, from soft tofu-like textures at low concentrations to cheese-like chewiness at higher concentrations. Unlike many conventional products, DF gels exhibit a uniquely aligned network, characterized by visually distinct lamellar domains (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003e) and hardness anisotropy ratios exceeding 1.1\u0026ndash;1.2. In structured foods, such anisotropic organization can enhance structural complexity, a key factor in creating diverse textural experiences and supporting product differentiation\u003csup\u003e\u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e27\u003c/span\u003e\u003c/sup\u003e. Freeze structuring thus provides a versatile approach for tailoring the texture and mouthfeel of legume-based gels across a wide range of culinary applications.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec6\" class=\"Section2\"\u003e \u003ch2\u003e3.4. Extending freeze-enabled texturization to diverse legume\u003c/h2\u003e \u003cp\u003eTo assess the broader applicability of freeze-induced structuring of minimally processed legumes, we extended the approach beyond chickpeas to a total of 96% of the legumes produced globally\u003csup\u003e\u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e28\u003c/span\u003e\u003c/sup\u003e (worldwide legume production, S14: Table S33). Gels made from whole red lentils (RL), green peas (GP), black beans (BB), mung bean (MB), black-eyed peas (BP), and soybeans (SB) were subjected to directional freezing and assessed for their structural alignment and textural properties. The resulting gels revealed apparent differences in textural stability and visual anisotropy (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eAll tested legumes formed anisotropically structured gels at 12.5 wt% solids, confirming the broad applicability of freeze-induced alignment (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003e). However, mechanical integrity varied among legume species. Red lentil (RL), black lentil (BL), and mung bean (MB) produced firm, stable gels with minimal syneresis, whereas black bean (BB) yielded slightly softer gels. Soybean (SB) gels at 12.5 wt% had low gel stability but still exhibited pronounced lamellar alignment under directional freezing. The textural properties are likely attributable to the elevated lipid and fibre content in soybeans, which can disrupt starch network formation and compromise overall gel integrity (SB composition, S16: Table\u0026nbsp;34). Increasing the solids concentration to 15.0, 17.5 or 20.0 wt% improved SB texture and retained structural anisotropy, indicating that mechanical properties can be fine-tuned by adjusting concentration. Furthermore, pH increase prior to heat treatment enhanced gel firmness at 12.5 wt%, with higher pH likely promoting protein unfolding, highlighting the role of protein solubility, particle size, and surface hydrophobicity in textural outcomes\u003csup\u003e\u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e29\u003c/span\u003e\u003c/sup\u003e. Other legumes, including black lentils, yellow lentils, kidney beans, navy beans, and lupins were also successfully anisotropically structured under DF (S15: Figure S7).\u003c/p\u003e \u003cp\u003eGel strength appeared to be influenced by the protein-to-starch (g/g) ratio of legumes (legume composition, S16: Table S34). Samples with relatively low ratios (e.g., red lentil 0.78, green pea 1.27, mung bean 0.61, black-eyed pea 0.70) formed self-supporting gels with higher mechanical integrity, likely due to sufficient starch availability for gelatinization and formation of a continuous load-bearing network. In contrast, samples with relatively high protein-to-starch ratios, such as soybean (3.39), exhibited weaker gel formation, likely due to limited starch content and protein dominance, which can hinder starch gelatinization and disrupt network continuity. Similar effects have been reported in mixed pea protein-maize starch systems, where increasing protein fractions weakened starch-dominated gel networks\u003csup\u003e\u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e30\u003c/span\u003e\u003c/sup\u003e. In addition to compositional ratios, starch-specific properties, including granule size, swelling capacity, and amylose to amylopectin ratio, may further modulate gel strength and network organisation, contributing to the observed variability among legumes\u003csup\u003e\u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e30\u003c/span\u003e,\u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e31\u003c/span\u003e\u003c/sup\u003e.\u003c/p\u003e \u003cp\u003eThese findings confirm that freeze structuring is broadly applicable across legumes and textural outcomes are strongly influenced by intrinsic composition. Key parameters such as solids concentration, particle size, and protein solubility can be strategically adjusted to tailor mechanical properties without compromising structural anisotropy. Further customization can be achieved by formulating mixtures of legumes with varying gel-forming capacities. Alternatively, process adaptations, such as modifying gelation, homogenization, or freezing conditions, offer further opportunities to fine-tune structure and texture. Freeze structuring enables the development of functional, anisotropic textures without reliance on refinement or purification steps, thereby preserving dietary fibre and other nutritionally and functionally valuable components. In contrast, conventional plant-based processing typically relies on fractionation and refinement to achieve functional products. For example, tofu production requires the separation of okara, resulting in substantial macronutrient redistribution\u003csup\u003e\u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e32\u003c/span\u003e\u003c/sup\u003e: approximately 85% of dietary fibre, 16% of lipids, and 23% of protein from whole soybeans are removed into okara and whey (compositions, S17, Table S35). Similarly, extrusion-based processes commonly involve ingredient fractionation to obtain functional protein or starch fractions\u003csup\u003e\u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e33\u003c/span\u003e\u003c/sup\u003e. These steps, although integral to those technologies, can result in nutrient losses and generate side streams\u003csup\u003e\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e,\u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e33\u003c/span\u003e\u003c/sup\u003e. By avoiding fractionation and refinement, freeze structuring retains the full nutritional profile of the raw material and supports more resource-efficient and sustainable food processing.\u003c/p\u003e \u003c/div\u003e"},{"header":"Conclusion","content":"\u003cp\u003eFreeze structuring provides a simple and robust strategy to generate anisotropic, fibrous textures in minimally processed whole-legume gels without the need for purification or additives. Directional freezing transforms legume suspensions into mechanically reinforced, anisotropic gels by redistributing macromolecular components and inducing lamellar architectures, as confirmed by microscopic, rheological, and mechanical analyses. Compared with conventional freezing or refrigeration, freeze-structured systems consistently exhibit higher mechanical strength and directional dependence of texture. The successful application across multiple legumes, including chickpeas, lentils, peas, and various beans, highlights the broad versatility of this approach and its capacity to produce diverse textural outcomes from a variety of raw materials.\u003c/p\u003e \u003cp\u003eBeyond its functional benefits, freeze structuring addresses key challenges in sustainable food processing. The method relies on simple freezing protocols that are compatible with existing cold-chain infrastructure and does not require specialized or processing steps beyond standard thermal treatment and freezing. At the household scale, the incremental energy demand of freezing is marginal in an already operating freezer, while at small- and medium-enterprise levels the process remains readily implementable using commonly available freezing systems. In contrast to conventional plant-based structuring technologies that rely on ingredient fractionation and refinement, often accompanied by nutrient losses and side-stream generation, freeze structuring operates directly on minimally processed legumes, thereby retaining dietary fibre and other valuable components. While prior research has primarily focused on purified protein or polysaccharide systems, this work demonstrates that directional freezing can effectively structure unrefined, thermally treated legume suspensions into fibrous gels with tunable mechanical properties. Together, these findings position freeze structuring as a globally relevant and scalable approach, adaptable to a wide range of legumes and processing contexts.\u003c/p\u003e \u003cp\u003eFuture translation toward consumer and culinary applications will benefit from the development of simple, reproducible recipes that integrate soaking, milling, thermal treatment, and freezing using standard kitchen equipment. The structuring mechanism remains effective in the presence of spices and herbs, enabling flavour customization without compromising texture. Providing clear procedural guidance alongside ingredient flexibility will support the adoption of freeze structuring in everyday practice and reinforce its potential contribution to sustainable, clean-label, and minimally processed food systems.\u003c/p\u003e"},{"header":"Methods","content":"\u003cp\u003eCommercial chickpeas (\u003cem\u003eCicer arietinum\u003c/em\u003e) were soaked in excess tap water for 24 h at 4\u0026deg;C. After soaking, the pulses were drained and diluted with water to achieve a dry matter content of 20.0% (w/w), then blended into a homogeneous slurry using a Vitamix Ascent A3500i blender (Vitamix, Ohio, USA). This stock was further diluted to working concentrations of 5.0, 7.5, 10.0, 12.5, 15.0, and 17.5% (w/w). Aliquots (50 mL) were transferred into Falcon tubes and heated in a water bath (Caso SV900 Sous Vide Gare, Caso Design, Arnsberg, Germany) at 90\u0026deg;C for 30 min. Samples were inverted every 30 seconds during the first 10 min and gently inverted every 5 min thereafter to ensure uniform thermal treatment. Three post-gelation conditions were directly applied: (1) Refrigeration (R): Samples were stored at 4\u0026deg;C for 25 hours in 50 mL falcon tubes. (2) Conventional Freezing (F): Samples were pre-cooled at 4\u0026deg;C for 1 h, then frozen in a shock freezer at -15\u0026deg;C for 24 h. (3) Directional freezing (DF): Samples were poured into custom insulated silicone moulds (setup, S2: Figure S2a and b), pre-cooled at 4\u0026deg;C for 1 h, and frozen unidirectionally at -15\u0026deg;C for 24 h using an aluminium plate as a cold source at the mould base. After freezing, all samples were thawed at room temperature for 4 h prior to analysis. The same procedure was performed on other legumes, including soybeans (\u003cem\u003eGlycine max\u003c/em\u003e), green peas (\u003cem\u003ePisum sativum)\u003c/em\u003e, black-eyed pea (\u003cem\u003eVigna unguiculata\u003c/em\u003e), red, black and yellow lentils (\u003cem\u003eLens culinaris\u003c/em\u003e), mung beans (\u003cem\u003eVigna radiata\u003c/em\u003e), lupins (\u003cem\u003eLupinus albus\u003c/em\u003e), and common beans (\u003cem\u003ePhaseolus vulgaris\u003c/em\u003e) namely black beans, kidney beans, navy beans.\u003c/p\u003e \u003cdiv id=\"Sec9\" class=\"Section2\"\u003e \u003ch2\u003e5.1. Rheology\u003c/h2\u003e \u003cp\u003eAmplitude sweeps were performed on chickpea gels prepared under the three structuring conditions (R, F, DF) using a rheometer (MCR 302, Anton Paar GmbH, Graz, Austria) with a plate-plate geometry (PP25), with both upper and lower plates roughened, and a gap of 1 mm at 25\u0026deg;C. Strain was increased from 0.01% to 300% at a constant frequency of 1 rad s⁻\u0026sup1;. From these sweeps, the plateau storage modulus (G\u0026prime;) and the limit of the linear viscoelastic region (LVER) was determined as the strain at which the storage modulus G' first dropped below 95% of its initial plateau value.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec10\" class=\"Section2\"\u003e \u003ch2\u003e5.2. Texture Analysis\u003c/h2\u003e \u003cp\u003eTexture was evaluated using single- and double-compression tests on a texture analyzer (TA.XTplus, Stable Micro Systems, Surrey, UK) equipped with the \u003cem\u003eExponent software\u003c/em\u003e (v6.2.6.0). Samples were equilibrated to room temperature and cut into cubes of 15 mm edge length using a sharp kitchen knife. R, F, and DF chickpea gels at 12.5, 15.0, 17.5, and 20.0 wt% were tested. Texture Profile Analysis (TPA) was performed with a 40 mm flat probe by compressing each sample twice with a 5 kg load cell, with DF measured parallel and perpendicular to the temperature gradient. Pre-test, test, and post-test speeds of 300, 180, and 300 mm min⁻\u0026sup1; were applied, respectively. Each compression reached 50% of the original sample height, with a 5 s interval between compressions. All measurements were performed in triplicate, using cubes taken from the middle section of each sample\u0026rsquo;s height. For reference values, the same procedure was used to test commercial Brie cheese, mozzarella, silken tofu, and firm tofu. Hardness was defined as the maximum force during the first compression, springiness as the ratio of sample height recovered between the first and second compression, cohesiveness as the ratio of the positive area of the second compression to that of the first compression, and chewiness as hardness \u0026times; cohesiveness \u0026times; springiness.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec11\" class=\"Section2\"\u003e \u003ch2\u003e5.3. Confocal Laser Scanning Microscopy (CLSM)\u003c/h2\u003e \u003cp\u003eR, F, and DF gels were cut into thin slices of approximately 1 mm using a razor blade. The CLSM method was adapted from Lyu et al. (2022)\u003csup\u003e\u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e30\u003c/span\u003e\u003c/sup\u003e. A staining solution containing fluorescein isothiocyanate isomer I (FITC, CAS 3326-32-7, Sigma-Aldrich, St. Louis, USA) and rhodamine B (CAS 81-88-9, Sigma-Aldrich, St. Louis, USA) (each at 0.0025 w/v%) in N,N-dimethylformamide (DMF, anhydrous, 99.8%, CAS 68-12-2, Sigma-Aldrich, Germany) was used. Gel slices were incubated in the dye solution for 1 hour, followed by three washes with deionized water over a total of 30 minutes to remove excess dye.\u003c/p\u003e \u003cp\u003eConfocal imaging was performed on a Leica TCS SP8 (Leica Microsystems, Wetzlar, Germany) using a 10\u0026times; objective (0.32 NA, HC PL FLUOTAR). Z-stacks were acquired for each sample (100 slices, 1 \u0026micro;m spacing) over a representative region. FITC was excited with a 488 nm laser and detected between 500\u0026ndash;530 nm. Rhodamine B was excited with a 552 nm laser and detected between 584\u0026ndash;620 nm. Images were adjusted for brightness and contrast.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec12\" class=\"Section2\"\u003e \u003ch2\u003e5.4. Photographic images\u003c/h2\u003e \u003cp\u003ePhotographic images were taking using a Canon EOS 70D (Canon Inc., Tokyo, Japan) mounted inside a photobox with constant lighting conditions. Images were adjusted for brightness and contrast.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec13\" class=\"Section2\"\u003e \u003ch2\u003e5.5. Digital Microscopy (DM)\u003c/h2\u003e \u003cp\u003eF, and DF gels were cut along the freezing temperature gradient into 3\u0026ndash;4 mm slices using a carpet knife and freeze-dried for 24 h in a freeze dryer (FreeZone 4.5 L, Labconco, Kansas City, USA) connected to a rotary vane vacuum pump (117 L min⁻\u0026sup1;, 230 V, Labconco, Kansas City, USA) at -84\u0026deg;C. For refrigerated gels, a slice of the same dimensions was immersed in liquid nitrogen and subsequently freeze-dried. Samples were imaged using a digital microscope (VHX-X1F, Keyence, Osaka, Japan) with a VH-Z20T lens and side-light adapter. Images were acquired at 100\u0026times; and 150\u0026times; magnification, stitched, and adjusted for brightness and contrast.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec14\" class=\"Section2\"\u003e \u003ch2\u003e5.6. Scanning Electron Microscopy (SEM)\u003c/h2\u003e \u003cp\u003eDF and F chickpea gels were cut across the freezing gradient into 3\u0026ndash;4 mm slices using a carpet knife and freeze-dried for 24 h in a freeze dryer (FreeZone 4.5 L, Labconco, Kansas City, USA) connected to a rotary vane vacuum pump (117 L min⁻\u0026sup1;, 230 V, Labconco, Kansas City, USA) at -84\u0026deg;C. For refrigerated gels, a slice of the same dimensions was immersed in liquid nitrogen and subsequently freeze-dried. Freeze-dried samples were trimmed into 4\u0026ndash;5 mm cubes and mounted on SEM stubs with the cut surface (perpendicular to the temperature gradient) facing upward. A 4 nm platinum coating was applied using a compact coating unit (CCU-010, Safematic GmbH, Switzerland). Imaging was performed on a GeminiSEM 450 (Carl Zeiss Microscopy GmbH, Jena, Germany) equipped with an SE2 detector, using an acceleration voltage of 2.00 kV and an aperture current of 100 pA.\u003c/p\u003e \u003c/div\u003e"},{"header":"Declarations","content":"\u003cp\u003eAcknowledgements\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eThe chair of Food Structure Engineering gratefully acknowledges financial support from Nestl\u0026eacute;, B\u0026uuml;hler, and Givaudan via the ETH Z\u0026uuml;rich Foundation.\u0026nbsp;The funders played no role in study design, data collection, analysis and interpretation of data, or the writing of this manuscript.\u003c/p\u003e\n\u003cp\u003eThe authors thank Marco Burkolter (Food Microbiology Group, ETH Z\u0026uuml;rich) for help with CLSM and Mathias Steinacher (Complex Materials Group, ETH Z\u0026uuml;rich) for his assistance in CLSM and SEM.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eAuthor information\u003c/p\u003e\n\u003cp\u003eAffiliation\u003c/p\u003e\n\u003cp\u003eDepartment of Health Sciences and Technology, Institute of Food, Nutrition and Health, ETH Z\u0026uuml;rich, Schmelzbergstrasse 7, Z\u0026uuml;rich, 8092, Z\u0026uuml;rich, Switzerland.\u003c/p\u003e\n\u003cp\u003eContribution\u003c/p\u003e\n\u003cp\u003eA.B., L.L., E.P., and P.R. conceptualized the work. A.B., E.P., L.L., and P.R. designed the methodology. A.B., E.P., and L.L. performed data collection. A.B., E.P., and P.R. wrote the original draft. A.B. created the figures. A.B., E.P., L.L. and P.R. revised the final version of the paper.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eCorresponding author(s)\u003c/p\u003e\n\u003cp\u003ePatrick A. R\u0026uuml;hs | [email protected]\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eCompeting interests\u003c/p\u003e\n\u003cp\u003eThe authors declare no competing interests.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n\u003cli\u003eRockstr\u0026ouml;m, J. \u003cem\u003eet al.\u003c/em\u003e The EAT\u0026ndash;Lancet Commission on healthy, sustainable, and just food systems. \u003cem\u003eThe Lancet\u003c/em\u003e 406, 1625\u0026ndash;1700 (2025).\u003c/li\u003e\n\u003cli\u003eCaputo, V., Sun, J., Staples, A. J. \u0026amp; Taylor, H. Market outlook for meat alternatives: challenges, opportunities, and new developments. \u003cem\u003eTrends Food Sci Technol\u003c/em\u003e 148, 104474 (2024).\u003c/li\u003e\n\u003cli\u003eFAO/OCCP. \u003cem\u003ePulses: Nutritious Seeds for a Sustainable Future\u003c/em\u003e. (FAO, 2016). doi:10.4060/I5528E.\u003c/li\u003e\n\u003cli\u003eVaz Patto, M. C. \u003cem\u003eet al.\u003c/em\u003e Achievements and challenges in improving the nutritional quality of food legumes. \u003cem\u003eCRC Crit Rev Plant Sci\u003c/em\u003e 34, 105\u0026ndash;143 (2015).\u003c/li\u003e\n\u003cli\u003eLisciani, S. \u003cem\u003eet al.\u003c/em\u003e Legumes and common beans in sustainable diets: nutritional quality, environmental benefits, spread and use in food preparations. \u003cem\u003eFront Nutr\u003c/em\u003e 11, 1385232 (2024).\u003c/li\u003e\n\u003cli\u003eHammer, L. \u003cem\u003eet al.\u003c/em\u003e Influence of processing on protein quality and environmental impact assessment of soy-based meat analogues. \u003cem\u003eFood Research International\u003c/em\u003e 222, 117636 (2025).\u003c/li\u003e\n\u003cli\u003eForde, C. G. Beyond ultra-processed: considering the future role of food processing in human health. \u003cem\u003eProceedings of the Nutrition Society\u003c/em\u003e 82, 406\u0026ndash;418 (2023).\u003c/li\u003e\n\u003cli\u003eGermerdonk, T., Bach, A., Marangoni, A. G., Mishra, K. \u0026amp; R\u0026uuml;hs, P. A. Unrefined plant raw materials are key to nutritious food. \u003cem\u003eNat Food\u003c/em\u003e 6, 657\u0026ndash;663 (2025).\u003c/li\u003e\n\u003cli\u003eDel Rio, A. R., Boom, R. M. \u0026amp; Janssen, A. E. M. Effect of fractionation and processing conditions on the digestibility of plant proteins as food ingredients. \u003cem\u003eFoods 2022, Vol. 11, Page 870\u003c/em\u003e 11, 870 (2022).\u003c/li\u003e\n\u003cli\u003eYee, C. S. \u003cem\u003eet al.\u003c/em\u003e Smart fermentation technologies: microbial process control in traditional fermented foods. \u003cem\u003eFermentation 2025, Vol. 11, Page 323\u003c/em\u003e 11, 323 (2025).\u003c/li\u003e\n\u003cli\u003eDe Henau, R., de Vries, A. \u0026amp; Rousseau, D. Structure and mechanical properties of anisotropic agar gels obtained via unidirectional freezing. \u003cem\u003eFood Research International\u003c/em\u003e 114626 (2024).\u003c/li\u003e\n\u003cli\u003eR\u0026uuml;hs, P., M\u0026uuml;ller, M. \u0026amp; Savorani, L. A. Freeze structured and enzymatically crosslinked food materials. (2024). US Patent 2024/0415147 A1\u003c/li\u003e\n\u003cli\u003eBryson, C., Rousseau, D., De Vries, A. \u0026amp; Gregson, C. M. Process for producing cookable, fibrous meat analogues with directional freezing. (2022). US Patent 11,241,024 B1\u003c/li\u003e\n\u003cli\u003eBasse, B., El Chemali, M. L., Dupuis, H. \u0026amp; Masbernat, L. Fibrous or laminated, and textured food product and method for producing same. (2024).\u003c/li\u003e\n\u003cli\u003eFischer, J., Bender, D., Domig, K. J. \u0026amp; Fuhrmann, P. L. Inducing anisotropy in emulsion-filled hydrogels by unidirectional freezing. \u003cem\u003eFood Hydrocoll\u003c/em\u003e 162, 111008 (2025).\u003c/li\u003e\n\u003cli\u003eNakagawa, K., Chantanuson, R., Boonarsa, P., Seephua, N. \u0026amp; Siriamornpun, S. Meat analogue preparation from cricket and rice powder mixtures with controlled textural and nutritional quality by freeze alignment technique. \u003cem\u003eFood Chem X\u003c/em\u003e 22, 101402 (2024).\u003c/li\u003e\n\u003cli\u003eRyu, J. \u0026amp; McClements, D. J. Freeze/thaw-triggered fixation of directionally frozen plant-based food matrices: Controlled release of gelling agents using double emulsions. \u003cem\u003eFood Hydrocoll\u003c/em\u003e 173, 112308 (2026).\u003c/li\u003e\n\u003cli\u003eSengar, A. S. \u003cem\u003eet al.\u003c/em\u003e Developing freeze-structured meat alternatives using pea and faba proteins: Evaluating their partial and complete substitution in beef patties. \u003cem\u003eFood Structure\u003c/em\u003e 45, 100451 (2025).\u003c/li\u003e\n\u003cli\u003eDurage, T. T. D. \u003cem\u003eet al.\u003c/em\u003e Developing a chickpea protein\u0026ndash;flaxseed oil emulsion gel meat analogue using the freeze-alignment technique. \u003cem\u003eSustainable Food Technology\u003c/em\u003e 3, 1996\u0026ndash;2008 (2025).\u003c/li\u003e\n\u003cli\u003eChantanuson, R., Nagamine, S., Kobayashi, T. \u0026amp; Nakagawa, K. Effect of dry heat treatment of soy protein powder on aligned structure formation in soy protein-based food gels during freezing. \u003cem\u003eJ Food Eng\u003c/em\u003e 363, 111779 (2024).\u003c/li\u003e\n\u003cli\u003eChantanuson, R., Nagamine, S., Kobayashi, T. \u0026amp; Nakagawa, K. Preparation of soy protein-based food gels and control of fibrous structure and rheological property by freezing. \u003cem\u003eFood Structure\u003c/em\u003e 32, 100258 (2022).\u003c/li\u003e\n\u003cli\u003eXu, L., Li, R. \u0026amp; Roe, B. Frozen food purchasing and home freezing of fresh foods: associations with household food waste. \u003cem\u003eBritish Food Journal\u003c/em\u003e 126, 4260\u0026ndash;4276 (2024).\u003c/li\u003e\n\u003cli\u003eBarbosa-C\u0026aacute;novas, G. V., Altunakar, B. \u0026amp; Mejia-Lorio, D. J. \u003cem\u003eFreezing of Fruits and Vegetables: An Agri-Business Alternative for Rural and Semi-Rural Areas\u003c/em\u003e (2005).\u003c/li\u003e\n\u003cli\u003eXiong, W., Devkota, L., Zhang, B., Muir, J. \u0026amp; Dhital, S. Intact cells: \u0026ldquo;Nutritional capsules\u0026rdquo; in plant foods. \u003cem\u003eCompr Rev Food Sci Food Saf\u003c/em\u003e 21, 1198\u0026ndash;1217 (2022).\u003c/li\u003e\n\u003cli\u003eZhang, C. \u003cem\u003eet al.\u003c/em\u003e Critical melting assisted freeze-thawing treatment as a novel clean-label way to prepare porous starch: Synergistic effect of melting and ice recrystallization. \u003cem\u003eFood Hydrocoll\u003c/em\u003e 131, 107730 (2022).\u003c/li\u003e\n\u003cli\u003eBach, A. \u0026amp; R\u0026uuml;hs, P. A. Addressing multifactorial complexity in freeze structuring of food colloids. \u003cem\u003eCurr Opin Colloid Interface Sci\u003c/em\u003e 101941 (2025).\u003c/li\u003e\n\u003cli\u003eOppen, D., Grossmann, L. \u0026amp; Weiss, J. Insights into characterizing and producing anisotropic food structures. \u003cem\u003eCrit Rev Food Sci Nutr\u003c/em\u003e 64, 1158\u0026ndash;1176 (2024).\u003c/li\u003e\n\u003cli\u003eFAO. FAOSTAT: Crops and livestock products. https://www.fao.org/faostat/en/#data/QCL. Licence: CC-BY-4.0 (2023).\u003c/li\u003e\n\u003cli\u003eSun, P. \u003cem\u003eet al.\u003c/em\u003e Improving gel properties of soy protein isolate through alkaline pH-shifting, mild heat treatment, and TGase cross-linking. \u003cem\u003eFood Hydrocoll\u003c/em\u003e 144, 108924 (2023).\u003c/li\u003e\n\u003cli\u003eLyu, Z., Sala, G. \u0026amp; Scholten, E. Water distribution in maize starch-pea protein gels as determined by a novel confocal laser scanning microscopy image analysis method and its effect on structural and mechanical properties of composite gels. \u003cem\u003eFood Hydrocoll\u003c/em\u003e 133, 107942 (2022).\u003c/li\u003e\n\u003cli\u003eCornejo-Ram\u0026iacute;rez, Y. I. \u003cem\u003eet al.\u003c/em\u003e The structural characteristics of starches and their functional properties. \u003cem\u003eCYTA - Journal of Food\u003c/em\u003e 16, 1003\u0026ndash;1017 (2018).\u003c/li\u003e\n\u003cli\u003evan der Riet, W. B., Wight, A. W., Cilliers, J. J. L. \u0026amp; Datel, J. M. Food chemical investigation of tofu and its byproduct okara. \u003cem\u003eFood Chem\u003c/em\u003e 34, 193\u0026ndash;202 (1989).\u003c/li\u003e\n\u003cli\u003eDel Rio, A. R., Boom, R. M. \u0026amp; Janssen, A. E. M. Effect of fractionation and processing conditions on the digestibility of plant proteins as food ingredients. \u003cem\u003eFoods 2022, Vol. 11, Page 870\u003c/em\u003e 11, 870 (2022). \u003c/li\u003e\n\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"npj-science-of-food","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"npjscifood","sideBox":"Learn more about [npj Science of Food](http://www.nature.com/npjscifood/)","snPcode":"41538","submissionUrl":"https://submission.springernature.com/new-submission/41538/3","title":"npj Science of Food","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"stoa","reportingPortfolio":"NPJ","inReviewEnabled":true,"inReviewRevisionsEnabled":true},"keywords":"Freeze structuring, legumes, plant-based, food, food structure, food texture","lastPublishedDoi":"10.21203/rs.3.rs-8663699/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-8663699/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eLegumes are sustainable, nutrient-rich, and affordable, yet their potential remains underused because conventional food processing often fails to utilize the entire raw material or achieve desirable textures. We show that freeze structuring can serve as a minimal-processing technique that imparts texture to legume-based foods without additives or raw material refinement. The method leverages directional ice crystal growth to concentrate starch, protein, and cell wall fragments into aligned interstitial regions as ice forms. Upon thawing, these concentrated domains interlink into a continuous network, reinforcing the gel and producing firmer textures with pronounced anisotropy. This approach was successfully applied to legumes representing 96% of global production, including chickpeas, soybeans, peas, lentils, lupins, common beans, and mung beans, underscoring its versatility. By eliminating fractionation and additive use, freeze structuring offers a decentralized, sustainable solution for producing nutritious, texturized plant-based foods using standard freezing technologies, accessible for industrial and home-scale production worldwide.\u003c/p\u003e\n\u003cp\u003e\u003cbr\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e*Andrea Bach and Elin Perler are co-first authors.\u003c/strong\u003e\u003c/p\u003e","manuscriptTitle":"Freeze-structuring unlocks minimal-processing strategies for legume texturization","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2026-02-13 13:18:34","doi":"10.21203/rs.3.rs-8663699/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Revision requested","date":"2026-02-28T12:52:00+00:00","index":"","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2026-02-27T10:12:34+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2026-02-27T04:37:55+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2026-02-26T01:27:28+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"331281370772730583229657217978709697350","date":"2026-02-08T14:26:50+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2026-02-08T11:40:17+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"205615265248382965918286486769369230395","date":"2026-02-08T08:44:04+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"96047770848253538205603456674534531724","date":"2026-02-08T04:30:46+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"209765662739731485557582610440070300722","date":"2026-02-08T04:22:12+00:00","index":"hide","fulltext":""},{"type":"reviewersInvited","content":"","date":"2026-02-08T04:19:53+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2026-01-26T03:58:48+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2026-01-26T03:58:41+00:00","index":"","fulltext":""},{"type":"submitted","content":"npj Science of Food","date":"2026-01-21T22:18:57+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"npj-science-of-food","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"npjscifood","sideBox":"Learn more about [npj Science of Food](http://www.nature.com/npjscifood/)","snPcode":"41538","submissionUrl":"https://submission.springernature.com/new-submission/41538/3","title":"npj Science of Food","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"stoa","reportingPortfolio":"NPJ","inReviewEnabled":true,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"201db56c-35cb-47fa-8bb3-fbd69e2dcd4e","owner":[],"postedDate":"February 13th, 2026","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"under-review","subjectAreas":[{"id":62659460,"name":"Physical sciences/Engineering"},{"id":62659461,"name":"Physical sciences/Materials science"},{"id":62659462,"name":"Biological sciences/Plant sciences"}],"tags":[],"updatedAt":"2026-04-19T11:08:17+00:00","versionOfRecord":[],"versionCreatedAt":"2026-02-13 13:18:34","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-8663699","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-8663699","identity":"rs-8663699","version":["v1"]},"buildId":"XKTyCvWXoU3ODBz1xrDgd","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}