Gelation behavior and mechanism of low methoxyl pectin in the presence of erythritol and sucrose: The role of co-solutes.

Co-solutes such as sucrose and sugar alcohol play a significant part in low methoxyl pectin (LMP) gelation. To explore their gelation mechanism, we investigated the gelation behavior of LMP in the presence of erythritol and sucrose with Ca2+. Results revealed that the introduction of erythritol and sucrose improved the hardness of the gels, fixed more free water, accelerated the rate of gel structuring, and enhanced the gel strength. FT-IR confirmed the reinforced hydrogen bonding and hydrophobic forces between the pectin chains after introducing co-solutes. And it could be observed clearly by SEM that the cross-linking density gel network enhanced with co-solutes. Furthermore, gel disruption experiments suggested the presence of ionic interaction, hydrogen bonding, and hydrophobic forces in LMP gels. Finally, we concluded that the egg-box regions cross-linked only by LMP and Ca2+ were too weak to form a stable gel network structure. Adding co-solutes could increase the amount of cross-linking between pectin chains and enlarge the cross-linking zones, which favored the formation of a dense gel network by more hydrogen bonding and hydrophobic forces. Sucrose gels had superior physicochemical properties and microstructure than erythritol gels due to excellent hydration capacity and chemical structure characteristics.

Effect of enzymatic de-esterification and RG-I degradation of high methoxyl pectin (HMP) on sugar-acid gel properties.

The influence of RG-I domains on high methoxyl pectin (HMP) sugar-acid gel properties has rarely been reported. In our work, HMP was modified by enzymatic de-esterification and degradation of RG-I domains to compare and analyze the relationship between the structure and final sugar-acid gel properties. The results showed that the degree of esterification (DE) of REP (pectin degraded by rhamnosidase) and GEP (pectin debranched by galactosidase) was the same as that of untreated HMP, whereas the DE of PMEP (pectin de-esterified by pectin methyl esterase) decreased from 59.63 % to 54.69 %. The monosaccharide composition suggested no significant changes in the HG and RG-I structural domains of PMEP. In contrast, the percentage of RG-I structural domains of REP and GEP dropped from 37 % to about 28 %, accompanied by a reduction in the proportion of the RG-I backbones and side chains. The rheological characterization of sugar-acid gels demonstrated an enhanced gel grade for PMEP and a weakened one for REP and GEP. Moreover, we constructed a correlation relationship between the fine structure of pectin and the properties of the sugar-acid gels, confirming the critical contribution of the RG-I region (especially the neutral sugar side chains) to the HMP sugar-acid gels.

Pectin gels based on H+/(NH4)2SO4 and its potential in sustained release of NH4.

A gelling strategy for HP was proposed in this study, ammonium sulfate (AS) as a co-solute could induce the gelling of HP in acidic environment. The solubility and Zeta potential of HP dramatically decreased in AS solution, which indicated AS could promote the aggregation of HP. The rheological results confirmed the gelling of HP (G' > G″) with AS: 25-30 wt% and pH ≤ 3.0, and the gel strength is mainly depended on HP rather than AS concentration. Smaller AS crystals (SEM) and reduced T2 values (LF-NMR) were observed in HP gels, suggested the gel network of HP could limit the migration of AS and water. Finally, it was found that the release process of NH4+ in HP + AS gel was lagged behind that of pure AS, which verified the potential of HP + AS gel in the field of sustained-release fertilizers.

Physicochemical characteristics and functional properties of high methoxyl pectin with different degree of esterification.

Moderate alkali de-esterification can change the physicochemical characteristics and thus the functional properties of high methoxyl pectin (HMP). The results revealed that de-esterification could increase negative charges (Zeta potential from -21 to -31 mV), decrease molecular weight (from 448 to 136 kDa) and apparent viscosity of HMP. Homogalacturonan (HG) content decreased (from 62% to 49%) while rhamnogalacturonan Ⅰ (RG-Ⅰ) content increased (from 32% to 46%) after de-esterification. The group characteristics of HMP with different degree of esterification (DE) were similar and no obvious impact was made on degree of crystallinity by alkali de-esterification. A conformation transition of HMP molecule implied by Congo red test were occurred as the DE decreased. With the decrease of DE, the molecular structure of HMP became shorter and smaller, and the entanglement was weaker. The de-esterification caused slight decrease of thermal stability. Alkali de-esterification would weaken the gel property and the emulsifying ability of HMP.

Citrus fiber for the stabilization of O/W emulsion through combination of Pickering effect and fiber-based network.

In this study, oil-in-water emulsions stabilized by citrus fiber were prepared and characterized. We found that citrus fiber can produce stable gel-like, surfactant-free O/W emulsions with microscale droplet sizes at fiber concentrations upon 2% (W/V) using 25% (V/V) oil. The interfacial framework, citrus fiber partition between the continuous phase and state of the droplets of emulsions were visualized by confocal laser scanning microscopy (CLSM), confirming that in addition to Pickering stabilization, the citrus fiber-based network also contributed to stabilization of the emulsions. The citrus fiber-stabilized emulsion is typical non-Newtonian fluid and its interfacial viscosity is not influenced obviously by changing pH from 2 to 10, ionic strength of NaCl from 0.00 to 1.00 mol/L or temperature from -20 to 70 °C. The acquired findings in this study show that citrus fiber can fabricate Pickering emulsions with excellent stability and solve the problem of resource waste during the pectin produce process.

Structural characterization of pectin-bismuth complexes and their aggregation in acidic conditions.

Bismuth-contained therapies are effective in treating gastric ulcer and eliminating Helicobacter pylori (Hp). Anion polysaccharides ligand could reduce the intake of bismuth, and enhance drug efficacy of bismuth compounds. In this study, pectin-bismuth (PB) was prepared and the changes of PB structure in acidic environment were reported for the first time. The structure of PB was characterized by FT-IR, XRD, and TGA, which suggested that combined with bismuth could alter the crystal structure of pectin. XPS confirmed the ionic binding of Bi3+ with carboxyl groups of pectin. The aggregating of PB with different pH level were also investigated, and the influence of pH on PB structure were observed by SEM. Results showed that PB has much larger volume of flocculation in acidic environment compared with bismuth nitrate. Additionally, apparent shear stress (τa) of PB suspension was evaluated. These results revealed the structural characteristics and acid-induced aggregation of pectin-bismuth, and bismuth could aggregate in acidic solution with the gelation of pectin.

Properties of dietary fiber from citrus obtained through alkaline hydrogen peroxide treatment and homogenization treatment.

In this research, citrus dietary fiber (CF) was modified by two methods, homogenization or alkalinehydrogenperoxide (chemical) treatment, to improve its physicochemical properties. The homogenization and chemical treatment highly increased the CF's water swelling capacity by 433% and 276%, respectively. The water holding capacity of CF significantly increased by 253% after homogenization and 197% after chemical treatment. Both treatments increased CF's total dietary fiber content and thermal stability. Moreover, the chemically treated CF thermal stability was higherthan the homogenized one. Scanning electron microscopy (SEM) results showed the modified CFs exhibited porous structure. XRD and NMR results showed that the CF's crystalline region could be disrupted by both treatments. Overall results suggest that the two treatments could effectivelyimprove CF physicochemical properties. The modified fibers might be potentially used as functional food ingredients.

Emulsifying properties of high methoxyl pectins in binary systems of water-ethanol.

Pectin is widely distributed in plant cell wall, most of which have limited emulsifying properties. Ethanol could alter the solubility of pectin, and affect its emulsifying properties. No creaming and breaking of emulsion appeared in 21% (v/v) ethanol contained emulsion. This project investigated the influence of ethanol (0%-35%, v/v) on conformation and emulsifying properties of pectin. Results shown that ethanol could reduce the helix conformation and zeta potential of pectin chain, which leading to compact conformation and enhanced interaction among pectin molecules. Although emulsion droplet diameter increased with ethanol content, the most stable emulsion was found in 21% (v/v) ethanol. CLSM also indicated over-aggregated pectin have a poor adsorption capacity on the interface of O/W. All results manifested the emulsifying properties of pectin can be improved by 21% (v/v) ethanol. This study provides a new strategy to improve the emulsifying property of pectin by changing its conformation.

Pea soluble polysaccharides obtained from two enzyme-assisted extraction methods and their application as acidified milk drinks stabilizers.

The objective of this work was characterize and evaluate the protein-stabilizing property of pea soluble polysaccharide (PSPS) extracted from pea by-products using spray-drying and ethanol precipitation oven drying, obtaining PSPS-A and PSPS-B, respectively. The weight average molecular weight (Mw) of PSPS-A and PSPS-B were 625 kDa and 809 kDa, respectively. The results of Fourier transform infrared spectroscopy (FT-IR) analysis indicated that PSPS-A, PSPS-B and soybean soluble polysaccharide (SSPS) contained the same functional groups. The absolute negative charges of PSPS-A or PSPS-B in aqueous solution were slightly higher than that of SSPS at pH 2.0 to 7.0. The apparent diameter of PSPS-B (479.1 nm) was larger than that of PSPS-A (127.7 nm) and SSPS (209.5 nm) were measured by dynamic light scattering. The AFM images revealed that both PSPS-A and PSPS-B possessed star-like structures with more side chains as compared to SSPS. It was found that the addition of 0.15% PSPS-A or 0.1% PSPS-B was adequate to prevent the aggregation of protein and obtain stable dispersion. Furthermore, PSPS has a wider pH range (pH 3.6-4.6) to stabilize milk protein than SSPS (pH 3.6-4.2).

Subcritical Water Induced Complexation of Soy Protein and Rutin: Improved Interfacial Properties and Emulsion Stability.

Rutin is a common dietary flavonoid with important antioxidant and pharmacological activities. However, its application in the food industry is limited mainly because of its poor water solubility. The subcritical water (SW) treatment provides an efficient technique to solubilize and achieve the enrichment of rutin in soy protein isolate (SPI) by inducing their complexation. The physicochemical, interfacial, and emulsifying properties of the complex were investigated and compared to the mixtures. SW treatment had much enhanced rutin-combined capacity of SPI than that of conventional method, ascribing to the well-contacted for higher water solubility of rutin with stronger collision-induced hydrophobic interactions. Compared to the mixtures of rutin with proteins, the complex exhibited an excellent surface activity and improved the physical and oxidative stability of its stabilized emulsions. This improving effect could be attributed to the targeted accumulation of rutin at the oil-water interface accompanied by the adsorption of SPI resulting in the thicker interfacial layer, as evidenced by higher interfacial protein and rutin concentrations. This study provides a novel strategy for the design and enrichment of nanovehicle providing water-insoluble hydrophobic polyphenols for interfacial delivery in food emulsified systems.

Fabrication and characterization of Pickering emulsions and oil gels stabilized by highly charged zein/chitosan complex particles (ZCCPs).

Herein, we reported a facile method to fabricate ultra-stable, surfactant- and antimicrobial-free Pickering emulsions by designing and modulating emulsions' interfaces via zein/chitosan colloid particles (ZCCPs). Highly charged ZCCPs with neutral wettability were produced by a facile anti-solvent procedure. The ZCCPs were shown to be effective Pickering emulsifiers because the emulsions formed were highly resistant to coalescence over a 9-month storage period. The ZCCPs were adsorbed irreversibly at the interface during emulsification, forming a hybrid network framework in which zein particles were embedded within the chitosan network, yielding ultra-stable food-grade zein/chitosan colloid particles stabilized Pickering emulsions (ZCCPEs). Moreover, stable surfactant-free oil gels were obtained by a one-step freeze-drying process of the precursor ZCCPEs. This distinctive interfacial architecture accounted for the favourable physical performance, and potentially oxidative and microbial stability of the emulsions and/or oil gels. This work opens up a promising route via a food-grade Pickering emulsion-template approach to transform liquid oil into solid-like fats with zero trans-fat formation.

The influence of heat treatment on acid-tolerant emulsions prepared from acid soluble soy protein and soy soluble polysaccharide complexes.

This research presents a green procedure to prepare oil in water (O/W) emulsion from acid soluble soy protein (ASSP) and soy soluble polysaccharide (SSPS), a long-term stable nanoscale system for delivering the lipophilic components. The emulsion technique involved the preparation complexion using ASSP and SSPS by electrostatic and hydrophobic interactions as well as high pressure homogenization. The average diameter of the droplet of emulsions (fresh and heated) is 263±2nm. Such emulsions resulted in heating stable dispersions containing corn oil at the concentration of 20.0%, even at the pH around the isoelectric points of ASSP. After 90days storage at 4°C, the mean diameter of emulsions after heating at 80°C for 60min is 314±1nm compared with 341±3nm of emulsions unheated. The heat-stability of dispersions were affected by emulsion conditions, so the present research demonstrates the emulsion stability against heat treatment, ionic strength and pH change.

Multilevel structural responses of ß-conglycinin and glycinin under acidic or alkaline heat treatment.

The structural responses of soy protein isolate under various processing conditions are of practically important for broadening its functionalities. In this work, multilevel structural responses of ß-conglycinin and glycinin under acidic or alkaline heat treatment were investigated. Our results suggested that heat treatment under acidic (i.e., pH2.5) or alkaline (i.e., pH8.5) conditions induced multilevel structural responses of ß-conglycinin and glycinin: under acidic heat treatment, both ß-conglycinin and glycinin underwent hydrolysis and experienced disruption and reorganization in ordered secondary structure. This process was accompanied with changes in tertiary structure where previously buried regions were exposed to the aqueous phase to different extent. Small-angle x-ray scattering (SAXS) results indicated that the protein conformations evolved from globular ones to elongated ellipsoids, with a more remarkable elongation effect in glycinin than ß-conglycinin. Upon solvent evaporation, ß-conglycinin favored aggregation with a low "lateral to vertical ratio", glycinin tended to aggregate in a high "lateral to vertical ratio" style. In contrast, alkaline heat treatment did not induce hydrolysis but disturbed the secondary structure instead. The protein monomers maintained the globular conformation and assembled into irregular large aggregates during solvent evaporation. Under either treatment, glycinin responded more sensitively than ß-conglycinin at all structure levels. The observed multi-level structural responses can be used to guide the rational modification of soy protein isolate with controlled conformation.

Fractionation and characterization of soy ß-conglycinin-dextran conjugates via macromolecular crowding environment and dry heating.

Conjugates of ß-conglycinin and dextran were prepared by heating in solution under macromolecular crowding environment and dry-heating methods, and then fractionated by solubility at pH 4.8 and pH 6.5 and characterized. The results showed that the degree of glycation of the conjugates extracted from pH 4.8 were higher than the conjugates extracted from pH 6.5. Corresponding to the higher degree of glycation, it was supposed that the ß-conglycinin of groups 4.8 of macromolecular crowding environment was completely surrounded by the dextran molecular while that of groups 6.5 were encircled partially with a lower degree of glycation. Compared to ß-conglycinin, groups 4.8 demonstrated a decreasing surface hydrophobicity and sulfhydryl group content while groups 6.5 increased. The secondary structure of ß-conglycinin soluble at pH 4.8 after conjugating under macromolecular crowding environment tended to stretch out and the highly ordered structure turn to random structures. The differences between the extraction of pH 4.8 and pH 6.5 conjugated by dry-heating methods were not as remarkable as the difference between the extraction conjugated by macromolecular crowding environment.

Fabrication and Characterization of Stable Soy ß-Conglycinin-Dextran Core-Shell Nanogels Prepared via a Self-Assembly Approach at the Isoelectric Point.

The preparation of soy ß-conglycinin-dextran nanogels (∼90 nm) went through two stages, which are safe, facile, and green. First, amphiphilic graft copolymers were formed by dextran covalently attaching to ß-conglycinin via Maillard dry-heating reaction. Second, the synthesized conjugates were heated above the denaturation temperature at the isoelectric point (pH4.8) so as to assemble nanogels. The effects of pH, concentration, heating temperature, and time on the fabrication of nanogels were examined. The morphology study displayed that the nanogels exhibited spherical shape with core-shell structures, which was reconfirmed by zeta-potential investigation. Both circular dichroism spectra and surface hydrophobicity analyses indicated that the conformations of ß-conglycinin in the core of nanogels were changed, and the latter experiment further revealed that the hydrophobic groups of ß-conglycinin were exposed to the surface of protein. The nanogels were stable against various conditions and might be useful to deliver hydrophobic bioactive compounds.

Soy lipophilic protein nanoparticles as a novel delivery vehicle for conjugated linoleic acid.

Soy lipophilic protein nanoparticles (LPP), which present a novel delivery vehicle for conjugated linoleic acid (CLA), were fabricated by ultrasonication of the soy lipophilic protein (LP), which exhibits unique characteristics including a high loading capacity, oxidation protection and a sustained releasing profile in vitro for CLA. The CLA-loaded LPP exhibited a mean diameter of 170 ± 0.63 nm and a loading capacity of 26.3 ± 0.40% (w/w). A coating of sodium caseinate (SC) on the surface improved the colloidal stability of the CLA-loaded LPP. This encapsulation conferred protection against the oxidation of CLA, by which the head space-oxygen consumption and hydrogen peroxide value were obviously decreased in comparison with the SC-encapsulated CLA and CLA alone. The delivery system enables a sustained releasing profile of CLA in a simulated gastrointestinal tract (GIT). These findings illustrate that the LPP could act as an effective delivery device for CLA, which could provide oxidation stability and a sustained release property.

Formation of soy protein isolate-dextran conjugates by moderate Maillard reaction in macromolecular crowding conditions.

BACKGROUND: Several methods have been reported for the conjugation of proteins with polysaccharides. Protein-polysaccharide conjugates can be formed by traditional dry heating, but this process is not attractive from an industrial viewpoint, and no commercial conjugates have been manufactured in this way. In the present study, in order to develop a more practical reaction method, macromolecular crowding was used to attach polysaccharides to proteins. RESULTS: Soy protein isolate-dextran conjugates (SDCs) were prepared via the initial stage of the Maillard reaction in macromolecular crowding conditions. The impact of various processing conditions on the formation of SDCs was investigated. The optimal conditions chosen from the experiments were a soy protein isolate/dextran ratio of 1:1 (w/w), a pH of 6.5, a reaction temperature of 60 °C and a reaction time of 30 h. Circular dichroism spectroscopy showed that the secondary and tertiary structures of the conjugates were changed significantly. Structural flexibility increased, allowing better display of their functional characteristics. The conjugates had a composition with various sizes, especially macromolecules, according to gel permeation chromatography. Thermal analysis showed that the thermal stability of the conjugates was improved. CONCLUSION: The production of SDCs under macromolecular crowding conditions appears to be an effective and promising technique, representing an advance over classic protein glycosylation methods.

Adsorption and dilatational rheology of heat-treated soy protein at the oil-water interface: relationship to structural properties.

We evaluated the influence of heat treatment on interfacial properties (adsorption at the oil-water interface and dilatational rheology of interfacial layers) of soy protein isolate. The related structural properties of protein affecting these interfacial behaviors, including protein unfolding and aggregation, surface hydrophobicity, and the state of sulfhydryl group, were also investigated. The structural and interfacial properties of soy protein depended strongly on heating temperature (90 and 120 °C). Heat treatment at 90 °C induced an increase in surface hydrophobicity due to partial unfolding of protein, accompanied by the formation of aggregates linked by disulfide bond, and lower surface pressure at long-term adsorption and similar dynamic interfacial rheology were observed as compared to native protein. Contrastingly, heat treatment at 120 °C led to a higher surface activity of the protein and rapid development of intermolecular interactions in the adsorbed layer, as evidenced by a faster increase of surface pressure and dilatational modulus. The interfacial behaviors of this heated protein may be mainly associated with more flexible conformation and high free sulfhydryl group, even if some exposed hydrophobic groups are involved in the formation of aggregates. These results would be useful to better understand the structure dependence of protein interfacial behaviors and to expand utilization of heat-treated protein in the formulation and production of emulsions.

In vitro assessment of the bioaccessibility of fatty acids and tocopherol from soybean oil body emulsions stabilized with ι-carrageenan.

The present investigation aimed to expand the knowledge of the in vitro bioaccessibility of fatty acids and tocopherol from natural soybean oil body emulsions stabilized with different concentrations of ι-carrageenan. Several physicochemical parameters including proteolysis of the interfacial layer, interfacial composition, and microstructure were evaluated with regard to their impact on the bioaccessibility of fatty acids and tocopherol. Results from simulated human digestion in vitro indicated that the bioaccessibility of total fatty acids and tocopherol decreased (62.7-8.3 and 59.7-19.4%, respectively) with the increasing concentration of ι-carrageenan. During the in vitro digestion procedure, ι-carrageenan affected physicochemical properties of the emulsions, thereby controlling the release of fatty acids and tocopherol. These results suggested that soybean oil body emulsions stabilized with ι-carrageenan could provide natural emulsions in foods that were digested at a relatively slow rate, the important physiological consequence of which might be increasing satiety.

Growth kinetics of amyloid-like fibrils derived from individual subunits of soy ß-conglycinin.

The amyloid-like fibrillation of soy ß-conglycinin subunits (α, α', and ß) upon heating (0-20 h) at 85 °C and pH 2.0 was characterized using dynamic light scattering, circular dichroism (CD), binding to amyloid dyes (Thioflavin T and Congo red), and atomic force microscopy. The fibrillation of all three subunits was accompanied by progressive polypeptide hydrolysis. The hydrolysis behaviors, fibrillation kinetics, and morphologies of amyloid-like fibrils considerably varied among α, α', and ß subunits. Faster hydrolysis rates and special fragments were observed for the α and α' subunits compared to the ß subunit. However, the order of the fibrillation rate and capacity to form ß-sheets was α' > ß > α, as evidenced by CD and Thioflavin T data. Moreover, sequential growth of twisted screw-structure fibrils, leading to macroscopic fibrils with distinct morphological characteristics, was observed for ß-conglycinin and individual subunits. The different fibrillation kinetics and morphologies of α, α', and ß subunits appear to be associated with the differences in the amino acid composition and typical sequence of peptides. Besides, the disruption of ordered structure of fibrils occurred upon further heating (6-20 h) due to extensive hydrolysis. These results would suggest that all subunits are involved in the fibrillation of ß-conglycinin, following multiple steps including polypeptide hydrolysis, assembly to amyloid structure, and growth into macroscopic fibrils with a fibril shaving process.