Textural improvement of pea protein-based high-moisture extrudates with corn zein and rice starch.

High moisture extrusion allows the production of plant protein-based products, including meat analogues. Building upon our previous findings showing that zein mixed with rice starch provides the necessary textural properties to formulations, different pea protein-based formulations with varying amounts of zein and rice starch or wheat gluten (as control) were produced using high moisture extrusion and the rheological, textural, and microstructural characteristics were evaluated and associated with the secondary structure of proteins. Samples containing wheat gluten presented desirable rheological and mechanical properties in terms of texturization, which was evidenced by the generation of a layered and three-dimensional viscoelastic network. The addition of rice starch to zein significantly increased the viscoelasticity of the samples due to enhanced development of non-covalent interactions that led to higher and more stable ß-sheets content and to the formation of a fibrous and layered microstructure and a 3D network nearly like those obtained with gluten. The sole replacement of pea protein by zein was not enough to develop these desired characteristics, demonstrating the importance of the non-covalent interactions between rice starch and zein for the generation of these properties. Overall, zein and rice starch improved texturization of pea protein-based gluten-free analogues made by high moisture extrusion.

Alternansucrase as a key enabling tool of biotransformation from molecular features to applications: A review.

Alternansucrase (ASR), classified in GH70, produces unique α-glucans with alternating α-1,3 and α-1,6 glycosidic linkages in the backbone chain from renewable sucrose which is easily obtained from nature with low cost. ASR has synthesized many products with valuable functionalities that hold enormous commercial interest and promising applications. The influence of biocatalysis and fermentation parameters on the yields, and properties of products are critical for the propositions made to promote the enzyme application. Investigations on ASR have been compiled in the review to provide information on the enzyme, products and parameters. This review summarizes studies on the characteristics, conversion mechanism, products, and beneficial applications of ASR and exhibits structure-based technologies to improve enzyme activity, specificity, and thermostability for industrial applications. Finally, prospects for further development are also proposed for various ASR applications in food and other fields.

Effect of limited proteolysis and CaCl2 on the rheology, microstructure and in vitro digestibility of pea protein-carboxymethyl cellulose mixed gel.

Limited proteolysis, CaCl2 and carboxymethyl cellulose (CMC) have individually demonstrated ability to increase the gel strength of laboratory-extracted plant proteins. However, the syneresis effects of their combination on the gelling capacity of commercial plant protein remains unclear. This was investigated by measuring the rheological property, microstructure and protein-protein interactions of gels formed from Alcalase hydrolyzed or intact pea proteins in the presence of 0.1 % CMC and 0-25 mM CaCl2. Sodium dodecyl-sulfate polyacrylamide gel electrophoresis (SDS-PAGE) showed the molecular weight of pea protein in the mixture were < 15 kDa after hydrolysis. The hydrolysates showed higher intrinsic fluorescence intensity and lower surface hydrophobicity than the intact proteins. Rheology showed that the storage modulus (G') of hydrolyzed pea protein (PPH)-based gels sightly decreased compared to those of native proteins. 5-15 mM CaCl2 increased the G' for both PP and PPH-based gels and decreased the strain in the creep-recovery test. Scanning electron microscopy (SEM) showed the presence of smaller protein aggregates in the PPH-based gels compared to PP gels and the gel network became denser, and more compact and heterogenous in the presence of 15 and 25 mM CaCl2. The gel dissociation assay revealed that hydrophobic interactions and hydrogen bonds were the dominant forces to maintain the gel structure. In vitro digestion showed that the soluble protein content in PPH-based gels was 10 âˆ¼ 30 % higher compared to those of the PP counterpart. CaCl2 addition reduced protein digestibility with a concentration dependent behavior. The results obtained show contrasting effects of limited proteolysis and CaCl2 on the gelling capacity and digestibility of commercial pea proteins. These findings offer practical guidelines for developing pea protein-based food products with a balanced texture and protein nutrition through formulation and enzymatic pre-treatment.

Effect of extrusion process conditions on extrudates enriched with carotenoids encapsulated by different methods using gum arabic and vegetable fat as carriers.

Bioactive compounds into extruded foods enhance their nutritional value but they are heat and shear labile and prone to oxidation. This study was aimed to examine the impacts of distinct encapsulation methods on the stability of carotenoids under typical extrusion conditions. The study presents innovative encapsulation methods and investigates the protection efficacy of carotenoids degradation, as well as the effects on the physicochemical characteristics of carotenoid-rich products. Thus, spray drying, spray chilling, and their combination were compared based on their ability to protect carotenoids. Processing temperatures were 110 °C and 140 °C, and shear rates 500 and 2000 1/s. Carotenoid retention was determined, ß- and α-carotene retention ranged from 17 to 44 % and 18 to 48 %, respectively. Upon storage at room temperature, the carotenoid content was stable for 15 days, followed by a marked reduction after 30 days. Extrudates enriched microparticles produced by spray chilling and the combined methods exhibited higher carotenoid protection during storage. They also showed better quality attributes, notably bulk density, high water absorption index, color properties, and carotenoid retention. These findings suggest that encapsulation can protect carotenoids during extrusion, and the protection can be tailored to optimize the attributes of the final products.

Editorial on the Special Issue "Novel Gels for Food Product Development".

Recently gels have gained significant attention in the food industry due to their unique properties and potential applications [...].

A Plant-Based Animal Fat Analog Produced by an Emulsion Gel of Alginate and Pea Protein.

As the market for plant-based meat analogs grows, the development of plant-based animal fat analogs has become increasingly important. In this study, we propose an approach by developing a gelled emulsion based on sodium alginate, soybean oil (SO), and pea protein isolate. Formulations containing 15% to 70% (w/w) SO were successfully produced without phase inversion. The addition of more SO resulted in pre-gelled emulsions with a more elastic behavior. After the emulsion was gelled in the presence of calcium, the color of the gelled emulsion changed to light yellow, and the formulation containing 70% SO exhibited a color most similar to actual beef fat trimming. The lightness and yellowness values were greatly influenced by the concentrations of both SO and pea protein. Microscopic images revealed that pea protein formed an interfacial film around the oil droplets, and the oil was more tightly packed at higher oil concentrations. Differential scanning calorimetry showed that lipid crystallization of the gelled SO was influenced by the confinement of the alginate gelation, but the melting behavior was like that of free SO. FTIR spectrum analysis indicated a potential interaction between alginate and pea protein, but the functional groups of SO were unchanged. Under mild heating conditions, gelled SO exhibited an oil loss similar to that observed in actual beef trims. The developed product has the potential to mimic the appearance and slow-rendering melting attribute of real animal fat.

Fabrication of phytoglycogen-derived core-shell nanoparticles: Structure and characterizations.

The objective of this work was to investigate the fabrication of core-shell nanoparticles using phosphorylase-catalyzed chain extension of phytoglycogen, and to analyze the changes of structure and characterizations in detail. During the glucosylation reaction, the inorganic phosphate increased substantially up to 2.3 mg/mL in the initial 12 h, and then increased incrementally to 2.5 mg/mL at 24 h. The similar to trends was observed for increasing Mw and Rz over time, due to glucosyl transfers on the surface chain to form a corona around the phytoglycogen core with a larger size. Phosphorylase modification increases the percentages of longer chain fractions and the average chain length increased from degree of polymerization (DP) 11.6 to DP 48.2. The modified phytoglycogen exhibited the characteristic of B-type crystalline structure, indicating that the specific core-shell nanoparticle with inner amorphous nature and outer crystalline layer. The above results revealed that the potentiality of enzymatic chain elongation of phytoglycogen to design novel core-shell nanoparticle with tailor-made structure and functionality.

Biomimetic synthesis of maltodextrin-derived dendritic nanoparticle and its structural characterizations.

The maltodextrin-derived dendritic nanoparticle was fabricated using microbial branching enzyme and its structural characterizations were investigated. During biomimetic synthesis, molecular weight distribution of maltodextrin substrate with 6.8 × 104 g/mol shifted to the narrower and uniform distribution region with the larger molecular weight up to 6.3 × 106 g/mol (MD12). The enzyme-catalyzed product had the larger size, higher molecular density as well as higher percentage of α-1,6 linkage, accompanying by more chain accumulations of DP 6-12 and disappearance of DP > 24, suggesting the biosynthesized glucan dendrimer had a compact tighter branched structure. The interaction of molecular rotor CCVJ and local structure of dendrimer was monitored, displaying there was a higher intensity related with the numerous nano-pockets at the branch points of MD12. The maltodextrin-derived dendrimers had the single spherical particulate shape with the size range of 10-90 nm. The mathematical models were also established to reveal the chain structuring during enzymatic reaction. The above results showed that the biomimetic strategy for novel dendritic nanoparticle with controllable structure arising from branching enzyme treated maltodextrin, which would help to enlarge the panel of available dendrimer.

Microencapsulation of Carotenoid-Rich Extract from Guaraná Peels and Study of Microparticle Functionality through Incorporation into an Oatmeal Paste.

The peels of guaraná (Paullinia cupana) fruit contain abundant carotenoid content, which has demonstrated health benefits. However, these compounds are unstable in certain conditions, and their application into food products can be changed considering the processing parameters. This study aimed to encapsulate the carotenoid-rich extract from guaraná peels by spray drying (SD), characterize the microparticles, investigate their influence on the pasting properties of oatmeal paste, and evaluate the effects of temperature and shear on carotenoid stability during the preparation of this product. A rheometer with a pasting cell was used to simulate the extrusion conditions. Temperatures of 70, 80, and 90 °C and shear rates of 50 and 100 1/s were the parameters evaluated. Microparticles with a total carotenoid content between 40 and 96 µg/g were obtained. Over the storage period, carotenoid stability, particle size, color, moisture, and water activity varied according to the core:carrier material proportion used. Afterward, the formulation SD1:2 was selected to be incorporated in oatmeal, and the paste viscosity was influenced by the addition of this powder. ß-carotene retention was higher than that of lutein following the treatment. The less severe treatment involving a temperature of 70 °C and a shear rate of 50 1/s exhibited better retention of total carotenoids, regardless of whether the carotenoid-rich extract was encapsulated or non-encapsulated. In the other treatments, the thermomechanical stress significantly influenced the stability of the total carotenoid. These results suggest that the addition of encapsulated carotenoids to foods prepared at higher temperatures has the potential for the development of functional and stable products.

Impact of ultra-shear technology on quality attributes of model dairy-pea protein dispersions with different fat levels.

This study investigated the impact of ultra-shear technology (UST) processing on dairy-pea protein dispersions with different fat levels. Raw milk, skim milk, and cream, as well as model dispersions with combinations of dairy products and pea protein (i.e., raw milk with pea protein, skim milk with pea protein, and cream with pea protein) were employed as test samples. UST experiments were conducted at a pressure of 400 MPa and 70 °C shear valve exit temperature. The UST treatment increased the viscosity of the dispersions and the increases depended on the fat level. Dairy-pea protein dispersions from raw milk and skim milk were shear thinning and mathematically described by the power-law model defined by the consistency coefficient, K (Pa·sn) and the flow behavior index, n. UST treated cream + pea protein dispersions produced structures with gel-like characteristics. Microstructure and particle size analysis determined by laser scanning microscope revealed a reduction in particle size after UST treatment in raw milk + pea protein and skim milk + pea protein dispersions up to 7.55 and 8.30 µm, respectively. In contrast, the particle mean diameter of cream + pea protein dispersions increased up to 77.20 µm after the UST treatment. Thus, the effect of UST on the particle size and rheological behavior of the dispersions depended on the fat level. UST-treated dispersions were stable with no visible phase separation or sedimentation upon centrifugation at 4000×g for 30 min (4 °C). Heat treatment and freeze-thaw treatment of UST-treated samples showed stable blends immediately after the treatments, but subsequent centrifugation showed solid separation. Results from the study suggest that UST is a potential technology to produce stable dairy + pea protein liquids foods with different rheological characteristics for diverse applications.

Plant protein-based fibers: Fabrication, characterization, and potential food applications.

Proteins from plants have been considered as safer, healthier, and more sustainable resources than their animal counterparts. However, incomplete amino acid composition and relatively poor functionality limit their applications in foods. Structuring plant proteins to fibrous architectures enhances their physicochemical properties, which can favor various food applications. This review primarily focuses on fabrication of fibers from plant proteins via self-assembly, electrospinning, solution blow spinning, wet spinning, and high-temperature shear, as well as on several applications where such fibrous proteins assemble in quality foods. The changes of protein structure and protein-protein interactions during fiber production are discussed in detail, along with the effects of fabrication conditions and protein sources on the morphology and function of the fibers. Self-assembly requires proteolysis and subsequent peptide aggregation under specific conditions, which can be influenced by pH, salt and protein type. The spinning strategy is more scalable and produces uniformed fibers with larger length scales suitable for encapsulation, food packaging and sensor substrates. Significant progress has been made on high-temperature shear (including extrusion)-induced fibers responsible for desirable texture in meat analogues. Structuring plant proteins adds values for broadened food applications, but it remains challenging to keep processes cost-effective and environmentally friendly using food grade solvents.

Pectin as a natural agent for reinforcement of pea protein gel.

Pectin has been used as a gel strengthening agent, but its role in pea protein gels remains unclear. The present study investigated the effects of low-methyl pectin on the physicochemical and rheological properties of pea protein gels at neutral pH and elucidated underlying gelling mechanisms. Pectin increased the stability and viscosity of pea protein dispersions and induced the formation of large protein aggregates. After heating, the storage modulus of pea protein gel containing 0.5 % pectin increased by nearly six times compared to those without pectin, and the gel microstructures became more heterogeneous and compact. Molecular docking revealed that pectin interacted with proteins via mainly electrostatic and hydrogen-bonding interactions. Solvent extraction found that pectin increased the hydrogen bonding, hydrophobic interactions, and disulfide bonds of the gel systems, thus boosting their strength. The current work provided a practical simple approach to increasing the strength of pea protein gels at a neutral pH.

Encapsulation of Rich-Carotenoids Extract from Guaraná (Paullinia cupana) Byproduct by a Combination of Spray Drying and Spray Chilling.

Guaraná byproducts are rich in carotenoids, featuring strong antioxidant capacity and health-promoting benefits. However, these compounds are highly susceptible to oxidation and isomerization, which limits their applications in foods. This research aimed to encapsulate the carotenoid-rich extract from reddish guaraná peels by spray drying (SD), chilling (SC), and their combination (SDC) using gum arabic and vegetable fat as carriers. The carotenoid-rich extract was analyzed as a control, and the formulations were prepared with the following core-carrier ratios: SD20 (20:80), SD25 (25:75), SD33 (33:67), SC20 (20:80), SC30 (30:70), SC40 (40:60), SDC10 (10:90), and SDC20 (20:80). The physicochemical properties of the formed microparticles were characterized, and their storage stability was evaluated over 90 days. Water activity of microparticles formed during the SD process increased during storage, whereas those formed by SC and SDC processes showed no changes in water activity. The formed microparticles exhibited color variation and size increase over time. Carotenoid degradation of the microparticles was described by zero-order kinetics for most treatments. Considering the higher carotenoid content and its stability, the optimum formulation for each process was selected to further analysis. Scanning electron micrographs revealed the spherical shape and absence of cracks on the microparticle surface, as well as size heterogeneity. SD increased the stability to oxidation of the carotenoid-rich extract by at least 52-fold, SC by threefold, and SDC by 545-fold. Analysis of the thermophysical properties suggested that the carrier and the process of encapsulation influence the powder's thermal resistance. Water sorption data of the SDC microparticles depended on the blend of the carrier agents used in the process. Carotenoid encapsulation via an innovative combination of spray drying and spray chilling processes offers technological benefits, which could be applied as a promising alternative to protect valuable bioactive compounds.

The effects of whey protein fibrils on the linear and non-linear rheological properties of a gluten-free dough.

The increasing awareness of the celiac disease, an autoimmune disorder caused by the consumption of products containing gluten, has led to a growing interest in the development of gluten-free bakery products. In this study, whey protein fibrils (WPFs) were incorporated to mimic the fibrous network of gluten. The rheological properties and microstructure of the developed gluten-free doughs were evaluated and compared with gluten doughs. Protein fibrils were prepared by heating a whey protein isolate (WPI) solution at 80°C in an acidic environment with low salt concentration, and then the fibril lengths were adjusted by leveling up the solution pH to 3.5 and 7. The dimensions of the fibrils were measured by atomic force microscopy (AFM). Rice and potato starches were mixed with fibrils, WPI, gluten, or without protein, to form different doughs for further investigation. Shear tests, including stress sweep, frequency sweep, and creep recovery, were performed to study the viscoelastic properties of doughs under small or large deformation. The strain-hardening properties of doughs under biaxial extension were studied by the lubricated squeezing flow method. The microstructure of the doughs was characterized by cryo-scanning electron microscopy (cryo-SEM). Compared with doughs prepared with WPI and no proteins, doughs incorporating fibrils showed comparable linear viscoelasticity to gluten dough tested with stress sweep, frequency sweep, and creep recovery in the linear viscoelastic region. More differences between the protein fibril doughs were revealed in the rheological properties in the non-linear region. Creep recovery parameters, such as compliance, elastic moduli during the creep, and recovery stages of gluten dough, were like those of WPF pH7 dough, but significantly different from those of the WPF pH3.5 dough. Strain-hardening properties were found in the WPF pH7 dough, although not in WPF pH3.5 dough. Microstructural characterization showed that both fibrils prepared with the different conditions formed a continuous protein phase for the improvement of dough cohesiveness, but the structure of the phase was different between the two fibrils. To summarize, whey protein fibril at pH 7 seemed to have the potential of being used as an ingredient with similar functions to gluten in gluten-free bakery products.

Influence of Hofmeister anions on structural and thermal properties of a starch-protein-lipid nanoparticle.

A self-assembled soluble nanoparticle, composed of common food biopolymers (carbohydrate, protein) and lipid, was previously reported by our laboratory. Although carrying capacity of valuable small molecules was demonstrated, physical functional properties are also important. Given the stabilization or destabilization characteristics of Hofmeister anion on macromolecular structures, mainly on proteins, here, we investigated the effects of different sodium salts composed of different Hofmeister anions on the structural and thermal properties of these self-assembled nanoparticles for improved functionalities. The salts were added into the mixture that was prepared in a diluted system during nanoparticle formation. Increased concentration of kosmotropic anions, in contrast to the chaotropic anion tested, resulted in nanoparticles with higher molar mass, hydrodynamic radius, and molecular density with more compact arrangement. The nanoparticles produced in presence of kosmotropic anions dissociated at higher temperatures and required higher enthalpies compared to the control sample. Spherical nanoparticles were formed for the kosmotropes with shear thinning behavior, contrary to rod-like nanoparticles for the chaotrope with near-Newtonian behavior. These findings help to gain an understanding of the effect of altering environmental conditions on the nanoparticles with an aim of producing desired structures for applications.

Rebuilding the lid region from conformational and dynamic features to engineering applications of lipase in foods: Current status and future prospects.

The applications of lipases in esterification, amidation, and transesterification have broadened their potential in the production of fine compounds with high cumulative values. Mostly, the catalytic triad of lipases is covered by either one or two mobile peptides called the "lid" that control the substrate channel to the catalytic center. The lid holds unique conformational allostery via interfacial activation to regulate the dynamics and catalytic functions of lipases, thereby highlighting its importance in redesigning these enzymes for industrial applications. The structural characteristic of lipase, the dynamics of lids, and the roles of lid in lipase catalysis were summarized, providing opportunities for rebuilding lid region by biotechniques (e.g., metagenomic technology and protein engineering) and enzyme immobilization. The review focused on the advantages and disadvantages of strategies rebuilding the lid region. The main shortcomings of biotechnologies on lid rebuilding were discussed such as negative effects on lipase (e.g., a decrease of activity). Additionally, the main shortcomings (e.g., enzyme desorption at high temperatre) in immobilization on hydrophobic supports via interfacial action were presented. Solutions to the mentioned problems were proposed by combinations of computational design with biotechnologies, and improvements of lipase immobilization (e.g., immobilization protocols and support design). Finally, the review provides future perspectives about designing hyperfunctional lipases as biocatalysts in the food industry based on lid conformation and dynamics.

Deciphering molecular interaction and digestibility in retrogradation of amylopectin gel networks.

The impact of the internal part of aewx amylopectin on the gel network and digestibility during retrogradation was investigated using wx amylopectin as a reference. After ß-amylolysis for 60 min (aewx-60), greater shifts in both λmax value and absorbance of iodine binding profiles were observed, accompanied by an increment of short chains (DP 3-5) with reducing the external long chains (DP 17.2). For the amylopectin gels aged 7 days at 4 °C, aewx had greater intermolecular aggregation of double helices to form junction zones, resulting in remarkably higher G', which was significantly greater than that of wx amylopectin or aewx-60. Moreover, aewx amylopectin had a greater RS accompanied by a reduction in RDS after retrogradation. The gel network models of retrograded amylopectins were built to interpret more molecular interactions for aewx than those of wx. The results revealed that aewx amylopectin with a higher proportion of longer external chains prompted the flexibility to align and interact for the formation of double helices and enzyme-resistant structures.

Atomistic Modeling of Peptide Aggregation and ß-Sheet Structuring in Corn Zein for Viscoelasticity.

The structure-function relationships of plant-based proteins that give rise to desirable texture attributes in order to mimic meat products are generally unknown. In particular, it is not clear how to engineer viscoelasticity to impart cohesiveness and proper mouthfeel; however, it is known that intermolecular ß-sheet structures have the potential to enhance the viscoelastic property. Here, we investigated the propensity of selected peptide segments within common corn α-zein variants to maintain stable aggregates and ß-sheet structures. Simulations on dimer systems showed that stability was influenced by the initial orientation and the presence of contiguous small hydrophobic residues. Simulations using eight-peptide ß-sheet oligomers revealed that peptide sequences without proline had higher levels of ß-sheet structuring. Additionally, we identified that sequences with a dimer hydrogen-bonding density of >22% tended to have a larger percent ß-sheet conformation. These results contribute to understanding how the viscoelasticity of zein can be increased for use in plant-based meat analogues.

Polyphenols Weaken Pea Protein Gel by Formation of Large Aggregates with Diminished Noncovalent Interactions.

Polyphenols are well-known native cross-linkers and gel strengthening agents for many animal proteins. However, their role in modifying plant protein gels remains unclear. In this study, multiple techniques were applied to unravel the influence of green tea polyphenols (GTP) on pea protein gels and the underlying mechanisms. We found that the elasticity and viscosity of pea protein gels decreased with increased GTP. The protein backbone became less rigid when GTP was present based on shortened T1ρH in relaxation solid-state NMR measurements. Electron microscopy and small-angle X-ray scattering showed that gels weakened by GTP possessed disrupted networks with the presence of large protein aggregates. Solvent extraction and molecular dynamic simulation revealed a reduction in hydrophobic interactions and hydrogen bonds among proteins in gels containing GTP. The current findings may be applicable to other plant proteins for greater control of gel structures in the presence of polyphenols, expanding their utilization in food and biomedical applications.

Structure and binding ability of self-assembled α-lactalbumin protein nanotubular gels.

Partial hydrolysis of whey-based α-lactalbumin (α-La) with Bacillus licheniformis protease (BLP) induces the formation of nanotubular structures in the presence of calcium ions by a self-assembly process. α-La nanotubes (α-LaNTs) exist in the form of regular hollow strands with well-defined average dimensions. The growth of nanotubes induces the formation of stiff transparent protein gels due to the well-arranged networks that the strands can form; these gels can be used for entrapment, transportation, and target delivery of bioactive agents in the industry. High purity of α-La (free of other whey protein fractions) is desirable for nanotube formation; however, pure proteins are very expensive and not practically obtained for industrial applications. Thus, the purpose of this research was to construct α-LaNTs from an α-La preparation with lower purity and to study the gelation phenomena triggered by the self-assembled nanotubes. Some structural features of nanotube gels and their active agent-binding abilities were also investigated. A lower amount of α-LaNTs was observed when low purity α-La was used for nanotube formation. Nanotube growth induced gel formation and higher gel stiffness was obtained when compared to α-La hydrolysates. α-La was denatured after hydrolysis and self-assembly, and remarkable changes were observed in the α-helix and ß-sheet domains of α-La structure. Increased intensity in Amide I and II regions indicated potential locations for binding of active agents to α-LaNTs. Whey-based α-La without much purification can be used to produce nanotubular gels and these gels can be considered carrying matrices for active agents in various industrial applications.