Transglutaminase Research Articles

Characteristics of whey protein concentrate/egg white protein composite film modified by transglutaminase and its application on cherry tomatoes.

In order to obtain food packaging film with better performance, whey protein concentrate (WPC) and egg white protein (EWP) were used as film-forming substrates, and its film properties weremodified by transglutaminase (TG). Then the effect of TG on the mechanical, physical, barrier, and microstructural properties of the WPC/EWP composite biodegradable film was investigated, and its preliminary application potential was explored. Compared toWPC and EWP films, WPC/EWP composite film had higher transmittance, tensile strength (TS), and thermal stability. Fluorescence results showed that the film experienced fluorescence quenching after TG treatment. Fourier transform infrared and x-ray diffraction results showed that WPC and EWP had good compatibility in the biodegradable film, the hydrogen bond interaction of film was increased due to TG, resulting in an increase in TS. Meanwhile, the water vapor permeability and contact angle of WPC/EWP film treated with TG at 5U/g protein increased by 28% and 76.1%, respectively. Besides, the WPC/EWP biodegradable film modified by TG (TG-W/E) was applied as a coating film on cherry tomatoes, effectively reducing the weight loss rate during storage from 14.2% to 10.8%. Furthermore, indexes, such as solid content, spoilage rate, hardness, pH, and lycopene, showed that the film had a good preservation effect on cherry tomatoes. To conclude, the appropriate addition of TG has a positive effect on the film properties of the WPC/EWP biodegradable film, which is beneficial to the development and utilization of protein-based film. WPC/EWP biodegradable film modified by TG has a great application prospect in extending the shelf life of fruit and vegetable.

Structural and physicochemical properties of plant-based meat analogues from transglutaminase-modified soybean protein

Plant-based meat analogues (PBMA) have been developed for decades. The resemblance in PBMA to meat, in terms of physical properties and sensory attributes, is important for consumers. This study aims to investigate the effects of transglutaminase (TG) concentration (1, 3, 5% (w/w)) on modified textured vegetable soybean protein (TVP). The changes in TVP protein molecular weight, microstructure, texture profile, sensory quality and nutritional value of PBMA were investigated. The first set of analyses examined the impact of TG concentration on TVP structure and the results showed that the TVP protein band from SDS-PAGE were changed to a larger band at 36-40 kDa as compared to the control indicating the formation of a cross-linked protein. The modified TVP was used as a raw ingredient for PBMA development. The appearance and microstructure of PBMA from the TG-modified TVP showed that the uncooked PBMA with TG-modified TVP were not different from those of the control. With regard to the texture profile of PBMA, an increase in TG concentration resulted in an increase in hardness. A sensory evaluation revealed that the PBMA with 1%TG provided the highest overall acceptance score. Hence, the PBMA with 1%TG was selected for a nutritional analysis comparing steak to meat. The carbohydrate and fat content of PBMA with 1%TG were higher than those of meat while the protein content of PBMA with 1%TG consisted of a maximum of 70% meat. This research could lead to alternative plant-based foods in the food industry.

Biomedical Application of Enzymatically Crosslinked Injectable Hydrogels.

Hydrogels have garnered significant interest in the biomedical field owing to their tissue-like properties and capability to incorporate various fillers. Among these, injectable hydrogels have been highlighted for their unique advantages, especially their minimally invasive administration mode for implantable use. These injectable hydrogels can be utilized in their pristine forms or as composites by integrating them with therapeutic filler materials. Given their primary application in implantable platforms, enzymatically crosslinked injectable hydrogels have been actively explored due to their excellent biocompatibility and easily controllable mechanical properties for the desired use. This review introduces the crosslinking mechanisms of such hydrogels, focusing on those mediated by horseradish peroxidase (HRP), transglutaminase (TG), and tyrosinase. Furthermore, several parameters and their relationships with the intrinsic properties of hydrogels are investigated. Subsequently, the representative biomedical applications of enzymatically crosslinked-injectable hydrogels are presented, including those for wound healing, preventing post-operative adhesion (POA), and hemostasis. Furthermore, hydrogel composites containing filler materials, such as therapeutic cells, proteins, and drugs, are analyzed. In conclusion, we examine the scientific challenges and directions for future developments in the field of enzymatically crosslinked-injectable hydrogels, focusing on material selection, intrinsic properties, and filler integration.

Elucidating the gas cell stabilization mechanism of buckwheat-wheat steamed bread induced by transglutaminase: A focus on the foaming and air-water interfacial properties of dough liquor

This work investigated the improvement mechanism of transglutaminase (TG) on the buckwheat-wheat steamed bread qualities from the view of protein structural, foaming and air-water interfacial properties of dough liquor (DL). TG reduced the hardness, ameliorated the crumb structure and enhanced the specific volume of buckwheat-wheat steamed bread by up to 9.0%. The DL yield, protein and water recovery were notably increased as the TG concentration increased to 0.2 U/g, then decreased with the higher TG concentrations. The low TG concentrations (<0.2 U/g) increased the protein molecular weight, and decreased the protein hydrophobicity, free amino and sulfhydryl groups content of DL. The high TG concentrations (>0.2 U/g) tended to induce both protein intermolecular and intramolecular cross-linking, which reduced the average particle size from 9.6 to 4.3 μm as the TG concentration increased from 0.2 to 1.0 U/g. TG markedly improved the foaming capacity and foam stability, decreased the surface tension and strengthened the interfacial films by enhancing the viscoelasticity modulus of buckwheat-wheat DL interfacial protein films. The topography of the air-water interfacial layer revealed that the TG-induced protein cross-linking led to a more homogeneous and denser topographical structure. This work will provide fresh insight into the effects of TG-induced protein cross-linking on gas cell stabilization and relate them to the buckwheat-wheat steamed bread quality properties.

Sodium caseinate/gellan gum emulsion gels for zeaxanthin dipalmitate delivery: Preparation, characterization, and gelation mechanism

The emulsion gel composed of proteins and polysaccharides exhibits significant potential as a targeted delivery system for bioactives in the gastrointestinal tract. In this study, a composite emulsion was prepared by using sodium caseinate (NaCas) and gellan gum (GG), which was subsequently crosslinked with transglutaminase (TG), glucono-δ-lactone (GDL), and calcium ions (Ca2+) to form emulsion gels. The physicochemical properties of the NaCas/GG emulsion gels were characterized to verify the effects of zeaxanthin dipalmitate encapsulation and protection, and a possible mechanism was discussed. The appearance and microscopy analyses revealed that the Ca2+-induced NaCas/GG emulsion gel exhibited superior shape retention and a denser network structure compared to GDL or TG crosslinking methods. The rheological analysis demonstrated that the Ca2+-crosslinked emulsion gels had higher modulus and hardness than their TG- or GDL- crosslinked counterparts. Infrared spectroscopy, molecular docking, and gelling force analysis revealed that the gelation mechanism of these emulsion gels primarily involved carbonyl-amide group crosslinking reactions, hydrogen bonding, and hydrophobic interactions. Furthermore, the Ca2+-crosslinked emulsion gel encapsulating zeaxanthin dipalmitate exhibited excellent thermal stability, resistance to gastrointestinal conditions, and stability. Overall, the above findings highlight that using Ca2+ crosslinking in NaCas/GG composite emulsion yields edible composite materials with enhanced functional performance, thereby expanding the possibilities for food gel design strategies.

Proteomic analysis revealing mechanisms of κ-carrageenan modulating the gelling properties of heat-induced microbial transglutaminase cross-linked myofibrillar protein gels

The effects of κ-carrageenan (KC, 0.2 %, w/w) on the gelling properties and microstructures of heat-treated myofibrillar protein (MP) gels mediated by various levels (0, 0.1, 0.3, and 0.5 %, w/w) of microbial transglutaminase (TG) were investigated. The results revealed that regardless of TG level, the existence of KC significantly promoted the forming of disulphide bonds but limited the generation of large-sized protein polymers in MP gel treated with TG alone, as well as subsequently enhanced the water retention ability and gel strength of mixed MP gel, especially in the sample adding 0.3% TG and 0.2% KC, which was clearly verified by the forming of more denser and continuous gel network. Moreover, TG-induced changes in protein structure led to the alteration in the protein profiles of different MP gels, and an obvious down-regulation trend of myosin (Q9TV63, Q9TV62, P79293, and Q9TV61) was identified in the TG-0.3 group. Furthermore, liquid chromatography–tandem mass spectrometry analysis demonstrated that TG mainly induces protein crosslinking in the light meromyosin domain of the myosin rod. Meanwhile, in the presence of KC, the crosslinking-site percentage of the TG-0.3 group decreased from 37.7% to 36.2%, indicating that KC potentially reduces TG-induced crosslinking in MP gels. Thus, our findings illustrate the critical mechanisms underlying the modulation of KC on the gelling properties of TG-mediated MP gels at the molecular level, which providing potential guideline for the combined applications of TG and KC in the meat industry.

Effect of transglutaminase on the structure, properties and oil absorption of wheat flour

The structure, properties, as well as the oil absorption characteristics of wheat flour (WF) treated with varying concentrations of transglutaminase (TG) (0 U/g ∼ 50 U/g) were characterized. The content of free amino groups in WF modified by TG (TG-WF) decreased and protein aggregated. The isopeptide bonds and disulfide bonds played important roles in protein crosslinking. The thermal stability, the peak viscosity after gelatinization and protein secondary structure stability of TG-WF were improved. In addition, the oil absorption and surface oil content of TG-WF after frying were reduced. TG enhanced the protein-protein interactions in WF, so that protein played barrier roles in the process of high-temperature frying, protecting the starch particles covered by them from the infiltration of oil, thus reducing the oil absorption of TG-WF during frying. Among them, the oil content of TG-WF-30 U/g after frying was the lowest, which decreased by 10.73 % compared with the control group.

Effect of polysaccharide on structures and gel properties of microgel particle reconstructed soybean protein isolate/polysaccharide complex emulsion gels as solid fat mimetic

In this work, a soybean protein isolate (SPI)/polysaccharide microgel particle reconstructed emulsion gels (MPEG) were fabricated through heat-induced gel (HG)–microgel particle–transglutaminase (TG) induced gel process in the presence of four polysaccharides (κ-carrageenan, κC; konjac glucomannan, KGM; high-acyl gellan, HA and xanthan gum, XG). HG exhibited a higher springiness than that of pig back fat (PBF) regardless of polysaccharide type and concentration. After forming MPEG, the springiness was significantly lowered at ≥0.6 % κC, which made MPEG exhibit similar springiness of PBF; while SPI/KGM, SPI/XG and SPI/HA systems failed to regulate the springiness property. Rheological behavior revealed the loss in elasticity, the increase in the plastic deformation of SPI/κC MPEG, while KGM, XG and HA systems still exhibited elasticity dominated rheological properties. Compared with KGM, XG, the presence of excess κC and HA disturbed the continuous protein network structure, resulting to the aggregation of microgel particles and oil droplets. Disulfide bonds and hydrophobic interactions mainly contributed to the formation of MPEG, while the addition of κC weakened the contribution of them, which was not conducive to the formation of gel network. This study provides a guidance on the development of solid fat mimetic based on the microgel particle emulsion gels.

Transglutaminase-catalyzed covalent anti-myostatin peptide depots

Nature realizes protein and peptide depots by catalyzing covalent bonds with the extracellular matrix (ECM) of tissues. We are translating this natural blueprint for the sustained delivery of a myostatin-inhibiting peptide (Anti-Myo), resulting in an enzyme depot established from injectable solutions. For that, we fused Anti-Myo to the D-domain of insulin-like growth factor I, a transglutaminase (TG) substrate. TG catalyzed the covalent binding of the D-domain to ECM proteins, such as laminin and fibronectin, on bioengineered ECM and in mice. ECM decorated with Anti-Myo suppressed myostatin activity and pathway activation and reduced the differentiation of preconditioned bone marrow-derived macrophages into osteoclasts in vitro.

Transglutaminase and protease: How starch granule-associated protein modification affects starch retrogradation

This study investigated the effects of transglutaminase (TG) and protease on starch granule-associated proteins and their impact on starch properties, especially retrogradation. TG incorporated storage protein onto the starch surface, while protease removed starch-granule associated proteins from it. Neither treatment altered the basic structural characteristics of starch such as its crystallinity or thermal properties, but could significantly affect swelling power, amylose leaching, and retrogradation. TG-treated wheat starch (WST) showed reduced swelling power (11.50) and amylose leaching (28.92 mg/g) compared with the control wheat starch (WSC) at 11.95 and 30.53 mg/g, respectively, suggesting a more stable structure. Conversely, protease-treated starch (WSP) exhibited increased swelling power at 12.47 and amylose leaching at 33.17 mg/g, suggesting a disintegration of starch granules. Both WSC and WST showed the same degree of retrogradation at the equilibrium state (a) at 0.06, whereas WSP, with a higher degree at 0.08, promoted retrogradation. The rate constant (k) for WST was lower (1.41 d−1) than that for WSC (2.37 d−1), indicating that TG treatment effectively inhibited retrogradation. These findings illustrated the distinct mechanisms through which TG and protease modify starch properties by acting on starch granule-associated proteins. This also highlighted the potential of TG and protease to control starch modifications for specific applications.

The impacts of different modification techniques on the gel properties and structural characteristics of fish gelatin

Fish gelatin hydrogels are typically characterized by weak texture and poor thermal stability. To address this issue, fish gelatin (FG) was modified using κ-carrageenan (κC) and transglutaminase (TG). FG-κC, FG-TG, and FG-κC-TG gels were prepared. The results demonstrated that the particle size and textural properties of FG gels were significantly enhanced by κC and κC-TG composite modifications, and their zeta potential was reduced (P < 0.05). As opposed to that, the improvement effects of TG modification were weaker. The highest particle size and hardness, along with the lowest zeta potential, were exhibited by the FG-κC-TG0.06% gel. Additionally, the FG-κC-TG gel system possessed higher apparent viscosity and elastic storage modulus. Fluorescence spectroscopy and UV absorption analysis indicated that fluorescence intensity was reduced by all three modification methods, and the conformational structure of FG was altered to varying degrees. Microstructure, FTIR, and XRD results revealed that the number of hydrogen bonds and isopeptide bonds in the composite gel systems was increased by κC and κC-TG modifications, enhancing the stability and orderliness of the gel's helical structure, and forming larger aggregates and denser network structures. Furthermore, due to the presence of covalent cross-linking, improved thermal stability was exhibited by the modified gelatin hydrogels. This study not only provides theoretical support for enhancing the performance of fish gelatin but also facilitates the application of modified fish gelatin in improving the sensory quality and flavor characteristics of food, as well as in serving as a matrix material in the biomedical field.

Impact of plant and animal proteins with transglutaminase on the gelation properties of clam Meretrix meretrix surimi

Different sources of proteins have various ways for facilitating the gelation process of surimi matrices, leading to the diverse combined effects with transglutaminase (TG). In this study, the effects of two plant proteins, soy protein isolate (SPI) and pea protein isolate (PPI), and two animal non-muscle proteins, egg white protein (EWP) and whey protein isolate (WPI), with TG addition on the gel properties of clam Meretrix meretrix (MM) surimi gels were investigated, and the mechanisms were explored. In the presence of TG, the exogenous protein addition significantly improved (p < 0.05) the gelation process of the MM surimi. The best improvements were obtained with the combination of EWP, where the gel strength, cooking loss, and water holding capacity were enhanced by 113.08%, 67.89%, and 26.30%, respectively. The addition of SPI showed a relatively weak enhancement compared to EWP incorporation, followed by the addition of PPI and WPI. The results of optical microscopy, differential scanning calorimetry, and chemical forces demonstrated that TG preferentially promoted plant proteins to form self-aggregation in the surimi system, and SPI generated more polymers than PPI. In contrast, the interpenetration of animal proteins and MM myofibrillar proteins was improved by TG. EWP formed a double cross-linking network within the MM protein network, while WPI acted as a binder to promote protein-protein interactions. This research could provide novel insights for developing MM resources and establish a theoretical foundation for the use of composite exogenous protein additives to enhance the gel properties of MM surimi products.

Association of CD24, CD27, and co-stimulatory molecules CD80 immunological marker expression on B-cells of human peripheral blood with development of celiac disease

Abstract: BACKGROUND: B-cells express a variety of clusters of differentiation markers during development. These markers determine the basic function of the immune phenotype specific to B-cells. Changes in the expression of these markers are linked to the development of many diseases, including chronic inflammation, autoimmune diseases, and immunodeficiency. OBJECTIVE: The current study aimed to investigate a change in CD24, CD27, and co-stimulatory molecules CD80 expression on peripheral blood B-cells and the extent of their contribution to celiac disease. MATERIALS AND METHODS: A total of 60 male children, whose ages ranged between 8 and 14 years, participated in this study. Thirty-five were identified as having celiac disease, while the control group comprised 25 children with anti-tissue transglutaminase (TG) (immunoglobulin G [IgG]), anti-tissue TG (IgA), and deamidated gliadin peptide (DGP) IgG levels normal. The frequency CD24, CD27, and CD80 expression were measured by flow cytometry. RESULTS: Celiac disease patients showed a substantial decrease in the percentage of CD24, CD27, and CD80 expression on B-cells compared to control groups. CONCLUSION: These findings suggest that numerical deficiency of CD24, CD27, and CD80 expression on B-cells in the peripheral blood mononuclear cell population, that may involve the loss of auto-tolerance that plays an important role in the immune response associated with inflammation and tissue damage in celiac disease. These immunological markers may be used as diagnostic indicators for this disease.

Preheating of Gelatin Improves its Printability with Transglutaminase in Direct Ink Writing 3D Printing.

Gelatin and transglutaminase (TG) ink is increasingly popular in direct ink writing three-dimensional (3D) printing of cellular scaffolds and edible materials. The use of enzymes to crosslink gelatin chains removes the needs for toxic crosslinkers and bypasses undesired side reactions due to the specificity of the enzymes. However, their application in 3D printing remains challenging primarily due to the rapid crosslinking that leads to the short duration of printable time. In this work, we propose the use of gelatin preheated for 7 days to extend the duration of the printing time of the gelatin ink. We first determined the stiffness of freshly prepared gelatin (FG) and preheated gelatin (PG) (5 – 20% w/w) containing 5% w/w TG. We selected gelatin hydrogels made from 7.5% w/w FG and 10% w/w PG that yielded similar stiffness for subsequent studies to determine the duration of the printable time. PG inks exhibited longer time required for gelation and a smaller increase in viscosity with time than FG inks of similar stiffness. Our study suggested the advantage to preheat gelatin to enhance the printability of the ink, which is essential for extrusion-based bioprinting and food printing.

Understanding the textural enhancement of low-salt myofibrillar protein gels filled with pea protein pre-emulsions through interfacial behavior: Effects of structural modification and oil phase polarity

This study investigated the effects of pea protein pre-emulsions containing triglyceride- or diglyceride-oil on the emulsifying and gelling properties of low-salt myofibrillar protein (MP). Pea protein isolates treated with pH12-shifting (PPIpH) or ultrasonication (PPIU) demonstrated superior initial interfacial adsorption and higher final interfacial pressure than native pea protein. Within MP/PPI blends, an increased ratio of MP led to a decrease in interfacial pressure, while simultaneously enhancing film elasticity at both polar and non-polar interfaces. Polar diglyceride promoted protein adsorption and fostered interfacial interactions between modified pea proteins and MP, enhancing the cross-linking of transglutaminase (TG) in the composite emulsion gels. Combining diglyceride-type PPIU and PPIpH emulsions with TG increased gel strength to 0.58 N and 0.63 N, respectively, from an initial 0.33 N, yielding a denser protein network with uniformly dispersed oil droplets. Therefore, the utilization of diglyceride and modified PPI can serve as structural enhancers in comminuted meat products.

Role of cuticular genes in the insect antimicrobial immune response.

Insects are established models for understanding host-pathogen interactions and innate immune mechanisms. The innate immune system in insects is highly efficient in recognizing and opposing pathogens that cause detrimental effects during infection. The cuticular layer which covers the superficial layer of the insect body participates in host defense and wound healing by inducing innate immune responses. Previous studies have started to address the involvement of cuticular genes in conferring resistance to insect pathogens, particularly those that infect by disrupting the insect cuticle. For example, the cuticular gene Transglutaminase (TG) in Drosophila melanogaster plays a structural role in cuticle formation and blood coagulation and also possesses immune properties against pathogenic infection. However, more information is becoming available about the immune function of other cuticular gene families in insects. In this review, we aim to highlight the recent advances in insect cuticular immunity and address the necessity of pursuing further research to fill the existing gaps in this important field of insect immunology. This information will lead to novel strategies for the efficient management of agricultural insect pests and vectors of plant and human disease.

Characteristics of the Mixed Yogurt Fermented from Cow-Soy Milk in the Presence of Transglutaminase.

The mixed yogurt was fermented from Cow-Soy milk and modified by transglutaminase (TG). The effects of mixed milk and TG on the quality characteristics of mixed yogurt were investigated by texture characteristics, rheology (rheometer) and structure (scanning electron microscopy). The findings revealed that the mixed yogurt with 50% cow milk exhibited lower hardness, viscosity and consistency. Furthermore, when TG was added, the yogurt showed better rheological properties, sensory score and a more stable microstructure. Compared with the samples without TG modification, the viscosity and cohesiveness of the modified samples increased by 10% and 100%, respectively. The combination of cow milk and soy milk improved the texture of yogurt, and the TG addition further improved the physicochemical properties of yogurt. This finding provided a meaningful reference for the development of mixed yogurt with a suitable taste from animal and plant milk, and laid a basis for the practical application of mixed yogurt in the dairy industry, which will meet the requirements for dairy products for consumers in future.

Optimizing skim milk yogurt properties: Combined impact of transglutaminase and protein-glutaminase

The lack of fat in yogurt can lead to alterations in taste and whey separation, reducing consumer acceptance. In this study, the feasibility of enhancing the quality of skim milk yogurt through a combination of transglutaminase (TG) and protein-glutaminase (PG) was investigated. The combination of TG and PG resulted in simultaneous cross-linking and deamidation of CN micelles, with PG deamidation taking priority over TG cross-linking, leading to higher solubility and lower turbidity of milk proteins compared with TG alone. When 0.06 U/mL TG and 0.03 U/mL PG were added, firmness and viscosity indexes significantly increased by 38.26% and 78.59%, respectively, as compared with the control. Microscopic images revealed increased cross-linking with CN and filling of cavities by smaller submicelles in the combination of TG and PG treatment. Furthermore, the combination of TG and PG resolved issues of rough taste and whey separation, leading to improved overall liking. This study highlights the benefits of using both enzymes in dairy production and has important implications for future research.

Effects of Laccase and Transglutaminase on the Physicochemical and Functional Properties of Hybrid Lupin and Whey Protein Powder.

Plant-based protein is considered a sustainable protein source and has increased in demand recently. However, products containing plant-based proteins require further modification to achieve the desired functionalities akin to those present in animal protein products. This study aimed to investigate the effects of enzymes as cross-linking reagents on the physicochemical and functional properties of hybrid plant- and animal-based proteins in which lupin and whey proteins were chosen as representatives, respectively. They were hybridised through enzymatic cross-linking using two laccases (laccase R, derived from Rhus vernicifera and laccase T, derived from Trametes versicolor) and transglutaminase (TG). The cross-linking experiments were conducted by mixing aqueous solutions of lupin flour and whey protein concentrate powder in a ratio of 1:1 of protein content under the conditions of pH 7, 40 °C for 20 h and in the presence of laccase T, laccase R, or TG. The cross-linked mixtures were freeze-dried, and the powders obtained were assessed for their cross-linking pattern, colour, charge distribution (ζ-potential), particle size, thermal stability, morphology, solubility, foaming and emulsifying properties, and total amino acid content. The findings showed that cross-linking with laccase R significantly improved the protein solubility, emulsion stability and foaming ability of the mixture, whereas these functionalities were lower in the TG-treated mixture due to extensive cross-linking. Furthermore, the mixture treated with laccase T turned brownish in colour and showed a decrease in total amino acid content which could be due to the enzyme's oxidative cross-linking mechanism. Also, the occurrence of cross-linking in the lupin and whey mixture was indicated by changes in other investigated parameters such as particle size, ζ-potential, etc., as compared to the control samples. The obtained results suggested that enzymatic cross-linking, depending on the type of enzyme used, could impact the physicochemical and functional properties of hybrid plant- and animal-based proteins, potentially influencing their applications in food.

Plant-based bigel based on chickpea-potato protein hydrogel and glycerol monostearate oleogel

The current research examines the effect of transglutaminase (TG) addition on novel bigel formulation based on a mixture of a chickpea-potato protein hydrogel and a glycerol monostearate (GMS)-oleogel processed using a hot emulsification procedure. Enzyme addition led to an increase in hardness and storage modulus values while similar melting behavior was found for both bigels with the main peak between 53–55 °C resulting from the GMS-oleogel melting. TGA thermograms of both bigels exhibited similar four weight-loss regions, with the control bigel having higher amount of bound water (∼56 %) compared to the TG-crosslinked bigel (∼50 %), which can be related to the higher stickiness of the control bigel. Confocal and cryo-HR-SEM imaging revealed the formation of a chickpea/potato protein network embedded with GMS oleogel droplets. Upon cooking, both bigels had crispy orange-brown surface with higher hardness and moderate browning due to the Maillard reaction.