3D printing of prawn mimics with faba proteins: The effects of transglutaminase and curdlan gum on texture.

With its capability for automated production of high-resolution structures, 3D printing can develop plant-based seafood mimics with comparable protein content. However, the challenge lies in solidifying 3D printed products to achieve the firmness of seafood. Targeting prawn, texturisation of its 3D printed mimic by curdlan gum was compared against incubation with a protein cross-linking enzyme, microbial transglutaminase. Faba bean protein extract (FBP) was selected for its lightest colour. To confer structural stability to the FBP-based ink without hindering extrudability, adding 1 % xanthan gum was optimal. Printed curdlan-containing mimics were steamed for 9 min, while printed transglutaminase-containing mimics were incubated at 55 °C before steaming. Either adding 0.0625 % or 0.125 % w/w curdlan or, incubating the transglutaminase-containing mimics for an hour achieved chewiness of 818.8-940.6 g, comparable to that of steamed prawn (953.13 g). Curdlan hydrogel penetrated and reinforced the FBP network as observed under confocal imaging. Whereas incubation of transglutaminase-containing mimics enhanced microstructural connectivity, attributable to transglutaminase-catalysed isopeptide cross-linkages, and the consequent increase in disulfide bonding and ß-sheet. Ultimately, transglutaminase treatment appeared more suitable than curdlan, as it yielded mimics with cutting strength comparable to steamed prawn. Both demonstrated promising potential to broaden the variety of 3D printed seafood mimics.

Pea protein isolate-soluble soybean polysaccharides electrostatic assembly: effect of pH, biopolymer mass ratio and heat treatment.

BACKGROUND: In past years, thousands of protein-polysaccharide complexes have been investigated to modify protein characteristics and functionality in food systems. However, the interaction between pea protein isolate (PPI) and soluble soybean polysaccharide (SSPS) has not been thoroughly characterized yet. RESULTS: In the present study, the phase behavior of PPI and SSPS mixtures was analyzed as a function of PPI:SSPS mixing ratio (1:1 to 1:0.10) and pH (7.0 to 2.0), showing that these biopolymers could be electrostatically assembled at 1:1 to 1:0.25 mixing ratios and 4.0 to 3.0 pH values. Then, the characteristics of the PPI-SSPS complexes were studied before and after heating (90 °C and 30 min) by ζ-potential, surface hydrophobicity, protein solubility, particle size distribution and physical stability for 56 days. By lowering the pH and PPI:SSPS mixing ratio, the complexes showed increased solubility, changed 𝜁-potential and higher physical stability. By heating, the complexes presented increased hydrophobicity and physical stability. CONCLUSION: Overall, PPI-SSPS complexes increased the protein solubility, reduced the particle size, and changed both the ζ-potential and the surface hydrophobicity with respect to PPI control, allowing stabilization of the colloidal system and broadening the possible applications of these high-quality proteins in acidic food systems. © 2024 Society of Chemical Industry.

Effects of apple fiber on the physicochemical properties and baking quality of frozen dough during frozen storage.

The effects of apple fiber on gluten structure and corresponding frozen dough quality during frozen storage were studied. The addition of 0.50% and 0.75% apple fiber effectively preserved gluten structure by inhibiting the breakage of disulfide bonds and promoting the formation of hydrogen bonds. Notably, the presence of 0.75% apple fiber increased the ß-turn of gluten from 29.60% to 33.84%. Fiber-enriched frozen dough exhibited a smoother and more compact microstructure, but excessive fiber addition (more than 1.00%) had adverse effects. The freezable water content of frozen dough decreased as fiber addition increased. Correspondingly, the addition of 1.50% apple fiber resulted in a 56.08% increase in storage modulus, indicating improved viscoelasticity of the dough. Consequently, the addition of 0.50% and 0.75% apple fiber alleviated the quality deterioration of frozen dough bread in terms of larger specific volume, softer and more uniform crumb.

Regulation of Intersubunit Interactions in Homotetramer of Glyceraldehyde-3-Phosphate Dehydrogenases upon Its Immobilization in Protein-Kappa-Carrageenan Gels.

Polysaccharides, being biocompatible and biodegradable polymers, are highly attractive as materials for protein delivery systems. However, protein-polysaccharide interactions may lead to protein structural transformation. In the current study, we analyze the structural adjustment of a homotetrameric protein, glyceraldehyde-3-phosphate dehydrogenase (GAPDH), upon its interactions with both flexible coil chain and the rigid helix of κ-carrageenan. FTIR spectroscopy was used to probe the secondary structures of both protein and polysaccharide. Electrostatically driven protein-polysaccharide interactions in dilute solutions resulted in an insoluble complex formation with a constant κ-carrageenan/GAPDH ratio of 0.2, which amounts to 75 disaccharide units per mole of protein tetramer. Upon interactions with both coiled and helical polysaccharides, a weakening of the intersubunit interactions was revealed and attributed to a partial GAPDH tetramer dissociation. In turn, protein distorted the helical conformation of κ-carrageenan when co-gelled. Molecular modeling showed the energy favorable interactions between κ-carrageenan and GAPDH at different levels of oligomerization. κ-Carrageenan binds in the region of the NAD-binding groove and the S-loop in OR contact, which may stabilize the OP dimers. The obtained results highlight the mutual conformational adjustment of oligomeric GAPDH and κ-carrageenan upon interaction and the stabilization of GAPDH's dissociated forms upon immobilization in polysaccharide gels.

Structure and rheological properties of extruded whey protein isolate: Impact of inulin.

Impacts of inulin addition (0, 5, 10, 15 %) on structure, functional and rheological properties of whey protein isolate (WPI) after extrusion pretreatment (E-WPI) were investigated. The results proved that after adding 15 % inulin, water holding capacity of gels, emulsifying activity, emulsion stability, foaming ability and foaming stability of E-WPI were the best and increased by 24.38 %, 7.43 %, 12.35 %, 162.97 % and 41.31 %, compared with those of unextruded WPI, respectively. Rheology analysis showed that apparent viscosity and consistency index of all the samples after inulin addition were enhanced and exhibited pseudoplastic fluids. FTIR spectroscopy indicated that E-WPI/WPI and inulin was linked together due to hydrogen bonds and addition of inulin increased the proportion of their ß-turn structure. These findings demonstrated that the addition of inulin in combination with extrusion pretreatment could jointly improve the functional properties of WPI. Therefore, E-WPI with the addition of inulin shows potential commercial applications in the production of novel food foaming agents and emulsifiers.

Impact of Flaxseed Gums on the Colloidal Changes and In Vitro Digestibility of Milk Proteins.

Flaxseed (Linum usitatissimum L.) mucilage is one of the most studied plant seed gums in terms of its techno-functional and health-promoting properties. Nonetheless, the interplay of flaxseed gum (FG) with other food biopolymers, such as milk proteins, under in vitro digestion conditions remains underexplored. The aim of the present work was to investigate the colloidal interplay between flaxseed gum (golden or brown) and milk proteins (sodium caseinate or whey protein isolate) under simulated in vitro digestion conditions and its relationship with the attained in vitro protein digestibility. The presence of flaxseed gum in the milk protein food models and in the oral food boluses obtained was associated with the occurrence of segregative microphase separation. Flaxseed gum exhibited a prominent role in controlling the acid-mediated protein aggregation phenomena, particularly in the sodium caseinate gastric chymes. The addition of FG in the food models was associated with a higher amount of intact total caseins and ß-lactoglobulin at the end of the gastric processing step. Monitoring of the intestinal processing step revealed a very advanced cleavage of the whey proteins (>98%) and caseins (>90%). The degree of the milk protein hydrolysis achieved at the end of the intestinal processing was significantly higher in the systems containing flaxseed gum (i.e., 59−62%) than their gum-free protein counterparts (i.e., 46−47%). It was postulated that the electrostatic milk protein complexation capacity and, to a lesser extent, the thickening effect of flaxseed gum influenced the in vitro digestibility of the milk proteins.

PEPscan: A Broad Spectrum Approach for the Characterization of Protein-Binder Interactions?

In a previous study, we have shown that PEPscan can provide a cheap and rapid means to identify candidate interfering peptides (IPs), i.e., peptides able to disrupt a target protein-protein interaction. PEPscan was shown to be effective in identifying a limited number of candidate IPs specific to the target interaction. Here, we investigate the results of 14 new PEPscan experiments for protein complexes of known 3D structures. We show that for almost all complexes, PEPscan is able to identify candidate IPs that are located at the protein-protein interface. The information it provides about the binding site seems, however, too ambiguous to be exploited in a simple manner to assist the modeling of protein complexes. Moreover, these candidates are associated with false positives. For these, we suggest they could correspond to non-specific binders, which leaves room for further optimization of the PEPscan protocol. Another unexpected advance comes from the observation of the applicability of PEPscan for polysaccharides and labeled peptides, suggesting that PEPscan could become a large spectrum approach to investigate protein-binder interactions, the binder not necessarily being a protein.

Property modelling of lysozyme-crosslinker-alginate complexes using latent variable methods.

Statistical methods were used to provide insight into a polymer complex system composed of lysozyme and alginate to quantify the effects of such parameters as pH, and ionic composition of the mixing solution on the properties of the complexes including composition, particle diameter, and zeta potential. Various crosslinkers (calcium, barium, iron[III], and bovine serum albumin), were used with lysozyme for complex formation to investigate the effect of crosslinker charge density on protein release kinetics, modelled using ktn . Multivariate statistical analysis showed that the kinetic parameters associated with the release were, not surprisingly highly dependent on the ionic strength of the release media, with higher ionic strength leading to faster release. The release parameter k was also shown to depend on the protein properties (size, ionic strength) while n was slightly, but not statistically dependent on the charge density of the crosslinker demonstrating that the nature of the crosslinker had minimal impact on drug release. The multivariate statistical has the potential to be used for optimization of the complexes and prediction of physical properties and degradation rates.

Newer guar gum ester/chicken feather keratin interact films for tissue engineering.

This work intends to synthesis newer guar gum indole acetate ester and design film scaffolds based on protein-polysaccharide interactions for tissue engineering applications. Guar gum indole acetate(GGIA) was synthesized for the first time from guar gum in presence of aprotic solvent activated hofmeister ions. The newer biopolymer was fully characterized in FT-IR,13C NMR, XRD and TGA analysis. High DS (Degree of Substitution, DS = 0.61) GGIA was cross-linked with hydrolyzed keratin, extracted from chicken feather wastes. Films were synthesized from different biopolymer ratios and the surface chemistry appeared interesting. Physicochemical properties for GGIA-keratin association were notable. Fully bio-based films were non-cytotoxic and exhibited excellent biocompatibility for human dermal fibroblast cell cultivations. The film scaffold showed 63% porosity and the recorded tensile strength at break was 6.4 MPa. Furthermore, the standardised film exerted superior antimicrobial activity against both the Gram-positive and Gram-negative bacteria. MICs were recorded at 130 µg/mL and 212 µg/mL for E. coli and S. aureus respectively. In summary, GGIA-keratin film scaffolds represented promising platforms for skin tissue engineering applications.

Synthesis of Nano-Scale Biopolymer Particles from Legume Protein Isolates and Carrageenan.

RESEARCH BACKGROUND: Food proteins and polysaccharides can be used for the synthesis of nano-scale biopolymer particles with potential applications in the fields of food and pharmaceuticals. This study focuses on utilizing legume proteins for the production of biopolymer particles via regulation of their electrostatic interactions with carrageenan. EXPERIMENTAL APPROACH: Protein isolates were obtained from mung bean (Vigna radiata), cowpea (Vigna unguiculata) and black gram (Vigna mungo) and their protein profiles were determined. Next, these isolates were allowed to interact with carrageenan at pH=5.0-7.0 to determine optimum conditions for obtaining nano-scale biopolymer particles. Selected biopolymer mixtures were then subjected to a heat treatment (85 °C for 20 min) to enhance the interactions among biopolymers. RESULTS AND CONCLUSION: Nano-scale biopolymer complexes were obtained at pH=6.5. They were roughly spherical in shape with a majority having a diameter in the range of approx. 100-150 nm. Heating of the biopolymer mixtures increased the diameter of the biopolymer particles by approx. 2.5-fold. In addition, their negative surface charge was increased, stabilizing them against aggregation over a broader pH range (4.0-7.0), enhancing their potential to be utilized in food matrices. NOVELTY AND SCIENTIFIC CONTRIBUTION: This study reports the applicability of mung bean, cowpea and black gram proteins for the synthesis of stable biopolymer particles. These biopolymer particles can be potentially used for the encapsulation and delivery of bioactive components.

Gelling and emulsifying properties of soy protein hydrolysates in the presence of a neutral polysaccharide.

Soy protein hydrolysates (SPH) with different degrees of hydrolysis (DH 0-16%) were obtained by varying the time of hydrolysis with bromelain. The objective of this study was to evaluate how selected techno-functional properties (gelation, emulsification) of SPH were affected by the presence of a non-gelling polysaccharide. A slight hydrolysis was beneficial to increase gel strength. Also, the emulsifying activity was improved for low DHs, whereas hydrolysis was detrimental for emulsion stability. Under certain conditions the presence of the non-gelling polysaccharide was beneficial to improve SPHs' functional properties, but the effect was in general complex and strongly dependent on both biopolymers' concentration and molecular weight. Nevertheless, it was demonstrated that by using SPH and galactomannan mixtures and controlling the biopolymers' concentration and molecular weight, improved functionalities can be obtained with useful applications in food formulation.

Use of gelatin and gum Arabic for encapsulation of black raspberry anthocyanins by complex coacervation.

Protein-polysaccharide interactions offer opportunities for designing the new functional foods with applications in the food and pharmaceutical industries. In this work, we microencapsulated black raspberry water extracts by double emulsion technique prior to complex coacervation using gelatin and gum Arabic to diminish the instability of their anthocyanins (ANCs) as water soluble compounds, particularly under harsh processing and storage conditions. Subsequently, optical microscopy, moisture, hygroscopicity, solubility, particle size, loading capacity, zeta potential, color parameters (L*, a*, b*, C, H° and TCD), stability, Fourier transform infrared spectroscopy (FTIR), and thermal behavior were evaluated. The microcapsules presented the lower moisture, hygroscopicity and solubility values as comparing the free form of ANCs. They showed the average sizes ranging from 35.34±3.21 to 80.22±5.21µm and high loading capacity (29.67±0.66-38.54±0.08%). According to the HPLC results, the selected method significantly increased the stability of ANCs up to 23.66% after 2 months of storage at 37±2°C. FT-IR spectra confirmed the negative zeta potential values of microcapsules and the occurrence of coacervation process. DSC results evidenced the thermostability of microcapsules. Furthermore, the selected optimal microcapsules revealed intense red color over the time of storage, implying the effectiveness of the method chosen to preserve anthocyanins.

Effect of the molecular weight of a neutral polysaccharide on soy protein gelation.

The effects of galactomannans with different molecular weights on the heat-induced gelation characteristics of soybean protein were investigated using dynamic small-strain rheometry, under conditions where the proteins carry a net negative charge (pH7). Microstructure of the resulting gels was investigated by confocal laser scanning microscopy. Phase-separated systems were obtained with different morphologies and degree of phase separation, depending on both biopolymer concentrations and polysaccharide molecular weight. In general, a gelling enhancing effect on soy proteins was verified, despite extensive phase-separation processes observed at the higher polysaccharide molecular weight. This effect was demonstrated by an increase of the gelation rate, a decrease in the temperature at the onset of gelation, and an increase of gel stiffness and elastic character, with the length of polysaccharide chains. Overall, the results obtained established that the judicious selection of the galactomannan molecular weight may be used to modify the structure and gelation properties of soy proteins, originating a diversity of rheological characteristics and microstructures that will impact on the design of novel food formulations.

Thermal gelation of mixed egg yolk/kappa-carrageenan dispersions.

This study aims to evaluate the effect of gum content and pH on the thermal gelation of mixed egg yolk/κ-carrageenan (EY/κC) dispersions, monitored by linear viscoelastic measurements. Heat processing induces dramatic changes in the microstructure and viscoelastic properties of EY/κC systems, which may be attributed to a multistage mechanism that yields an interparticle gel network. An increase in κC content generally induces an enhancement in viscoelasticity. A reduction in pH hinders this enhancement and causes an anticipation of the multistage process, which confirms the importance of the electrostatic interactions of EY/κC dispersions. The viscoelastic properties of EY/κC gels generally fit a master mechanical spectrum, which suggests that the protein matrix generally dominates the microstructure of EY/κC gels. However, SEM images reveal formation of a κC network at low pH, at which some κC autohydrolysis may also play a role. Electrostatic attractions seem to favour interactions among EY aggregates and κC into the carrageenan network.

Morphology of complexes formed between ß-lactoglobulin nanofibrils and pectins is influenced by the pH and structural characteristics of the pectins.

For the optimal use of ß-lactoglobulin nanofibrils as a raw material in biological composites an in-depth knowledge of their interactions with other constituents is necessary. To understand the effect of electrostatic interactions on the morphology of resulting complexes, ß-lactoglobulin nanofibrils were allowed to interact with pectins in which the amount of available negative charge was controlled by selecting their degree of methylesterification. In this study, citrus pectins having different degrees of methylesterification (∼48, 67, 86, and 97%) were selected and interacted with nanofibrils at pH 2 and pH 3, where they possess a net positive charge. Electrostatic complexes formed between ß-lactoglobulin nanofibrils and all pectin types, except for the sample having a degree of methylesterification of 97%. The morphology of these complexes, however, differed significantly with the degree of methylesterification of the pectin, its concentration, and the pH of the medium, revealing that distinct desired biological architectures can be attained relatively easily through manipulating the electrostatic interactions. Interestingly, the pectin with a degree of methylesterification of 86% was found to crosslink the ß-lactoglobulin nanofibrils into ordered 'nanotapes'.

Effect of Xanthan Gum on the Rheological Behavior and Microstructure of Sodium Caseinate Acid Gels.

The aim of this work was to study the effect of xanthan gum (XG) on the gelation process of bovine sodium caseinate (NaCAS) induced by acidification with glucono-δ-lactone (GDL) and on the mixed acid gel microstructure. Before GDL addition, segregative phase separation was observed in all the NaCAS-XG mixtures evaluated. The gelation process was analyzed by using a fractional factorial experimental design. The images of the microstructure of the mixed acid gels were obtained by conventional optical microscopy and the mean diameter of the interstices was determined. Both the elastic character and the microstructure of the gels depended on the concentrations of XG added. As XG concentration increased, the kinetics of the gelation process was modified and the degree of compactness and elasticity component of the gel network increased. The microstructure of gels depends on the balance among thermodynamic incompatibility, protein gelation and NaCAS-XG interactions.

Protein/polysaccharide interactions and their impact on haze formation in white wines.

Proteins in white wines may aggregate and form hazes at room temperature. This was previously shown to be related to pH-induced conformational changes and to occur for pH <3.5. The aim of the present work was to study the impact of wine polysaccharides on pH-induced haze formation by proteins but also the consequences of their interactions with these proteins on the colloidal stability of white wines. To this end, model systems and purified global pools of wine proteins and polysaccharides were used first. Kinetics of aggregation, proteins involved, and turbidities related to final hazes were monitored. To further identify the impact of each polysaccharide, fractions purified to homogeneity were used in a second phase. These included two neutral (mannoprotein and arabinogalactan) and two negatively charged (rhamnogalacturonan II dimer (RG-II) and arabinogalactan) polysaccharides. The impact of major wine polysaccharides on wine protein aggregation at room temperature was clearly less marked than those of the pH and the ionic strength. Polysaccharides modulated the aggregation kinetics and final haziness, indicating that they interfere with the aggregation process, but could not prevent it.

Characterisation of the interaction of neuropilin-1 with heparin and a heparan sulfate mimetic library of heparin-derived sugars.

Background. Neuropilin-1 (NRP-1) is a multidomain membrane protein with soluble isoforms interacting with a complex network of other membrane receptors, their respective ligands and heparan sulfate (HS). It is involved in the development of vasculature, neural patterning, immunological responses and pathological angiogenesis. Methods. We have characterised the binding of a Fc fusion of rat NRP-1 (Fc rNRP-1) and of a soluble isoform, corresponding to the first four extracellular domains of human NRP-1, shNRP-1, using optical biosensor-based binding assays with a library of heparin derivatives. Selective labelling of lysines protected upon heparin binding allowed their identification by mass spectrometry. Results. Fc rNRP-1 bound to heparin with high affinity (2.5 nM) and fast ka (9.8 × 10(6) M(-1)s(-1)). Unusually, NRP-1 bound both highly sulfated and completely desulfated stretches of heparin and exhibited a complex pattern of preferences for chemically modified heparins possessing one or two sulfate groups, e.g., it bound heparin with just a 6-O sulfate group better than heparin with any two of N-sulfate, 6-O sulfate and 2-O sulfate. Mass-spectrometry based mapping identified that, in addition to the expected the b1 domain, the a1, and c domains and the L2 linker were also involved in the interaction. In contrast, shNRP-1 bound heparin far more weakly. This could only be shown by affinity chromatography and by differential scanning fluorimetry. Discussion. The results suggest that the interaction of NRP-1 with HS is more complex than anticipated and involving a far greater extent of the protein than just the b1-b2 domains. NRP-1's preference for binding long saccharide structures suggests it has the potential to bind large segments of HS chains and so organise their local structure. In contrast, the four domain soluble isoform, shNRP-1 binds heparin weakly and so would be expected to diffuse away rapidly from the source cell.

Phase behavior of ovalbumin and carboxymethylcellulose composite system.

The phase behavior, rheological properties and microstructure of ovalbumin and carboxymethylcellulose (OVA-CMC) conjugates were studied and the influence parameters were investigated. The results showed that the phase behavior of OVA-CMC conjugates was related to pH and concentration of CMC and NaCl. When pH was over 5.0, discrete phase separation occurred in the mixture system, which indicated that OVA and CMC were thermodynamic incompatible. The mixture system turned into uniform stable emulsion system when pH reduced below 5.0. The addition of NaCl can improve the stability of composite system against pH sensitivity. CLSM and particle size distribution and ultraviolet spectrum analysis results confirm that emptying interactions play a leading role in the separation system.