高温短时干热法制备SPI-麦芽糊精糖基化产物及其乳化性的研究

采用高温(90,115和140℃)短时(2 h)干热法制备了大豆分离蛋白(SPI)-麦芽糊精(MD)糖基化产物。利用接枝度和SDS-PAGE研究了SPI与MD发生糖基化反应的程度。同时考察了SPI-MD糖基化产物稳定水包油乳液的性质,并探讨了盐离子和热处理对SPI-MD糖基化产物稳定乳液的储藏稳定性的影响。试验发现,提高反应温度能增大糖基化反应的接枝度,且pH 3.0时,与添加盐离子或经过热处理的SPI,SPI-MD糖基化产物(90℃)和SPI-MD糖基化产物(115℃)稳定乳液相比,添加盐离子或经过热处理的SPI-MD糖基化产物(140℃)的稳定的乳液具有较高的乳化稳定性和储藏稳定性。     

Preparation,characterisation and selected functional properties of hydrolysed sodium caseinate–maltodextrin conjugate

Sodium caseinate was hydrolysed to a limited, moderate or extensive degree. The hydrolysates were conjugated with maltodextrin by a Maillard‐type reaction by dry‐heat treatment at 60 °C and 79% relative humidity for 2 or 4 days. Conjugates were characterised by SDS–PAGE and gel permeation chromatography. In comparison with the hydrolysates themselves, the conjugated hydrolysates had improved solubility, particularly around the isoelectric pH of the protein. The emulsifying properties of these conjugates were assessed in oil‐in‐water (o/w) emulsions; on emulsion formation, each conjugate‐stabilised emulsion had lower mean fat globule size than the corresponding hydrolysate‐stabilised emulsion. After storage for 7 days under accelerated shelf life testing conditions, the limited and moderate hydrolysate conjugate–stabilised emulsions had improved storage stability compared with hydrolysate‐stabilised emulsions; however, further research is required to optimise the hydrolysate fraction prior to conjugation for the production of novel low molecular weight emulsifiers.     

Emulsification properties of sodium caseinate‐based conjugates with selected polysaccharides

Sodium caseinate (SC) was conjugated with polysaccharides, viz. maltodextrin (MD), pectin (P) and gum arabic (GA) at protein:polysaccharide weight ratio of 1:2, 1:1 and 2:1. The emulsifying properties and other relevant chemical properties of these conjugates were compared. The visible colour change, SDS‐PAGE analysis and available reducing groups confirmed greater conjugation in SC‐MD conjugate than the SC‐GA and SC‐P conjugates. SC‐P conjugate at the weight ratio of 1:2 exhibited the best emulsifying properties (emulsifying activity – 46.7%, emulsion stability – 7 days at 5 ± 1 °C storage) and had better solubility (33.5%) near the iso‐electric pH.     

Stability of oil‐in‐water emulsions as influenced by thermal treatment of whey protein dispersions or emulsions

Properties of whey protein concentrate stabilised emulsions were modified by protein and emulsion heat treatment (60–90 °C). All liquid emulsions were flocculated and the particle sizes showed bimodal size distributions. The state and surface properties of proteins and coexisting protein/aggregates in the system strongly determined the stability of heat‐modified whey protein concentrate stabilised emulsions. The whey protein particles of 122–342 nm that formed on protein heating enhanced the stability of highly concentrated emulsions. These particles stabilised protein‐heated emulsions in the way that is typical for Pickering emulsions. The emulsions heated at 80 and 90 °C gelled due to the aggregation of the protein‐coated oil droplets.     

Surface Properties of Heat‐Induced Soluble Soy Protein Aggregates of Different Molecular Masses

Suspensions (2% and 5%, w/v) of soy protein isolate (SPI) were heated at 80, 90, or 100 °C for different time periods to produce soluble aggregates of different molecular sizes to investigate the relationship between particle size and surface properties (emulsions and foams). Soluble aggregates generated in these model systems were characterized by gel permeation chromatography and sodium dodecyl sulfate‐polyacrylamide gel electrophoresis. Heat treatment increased surface hydrophobicity, induced SPI aggregation via hydrophobic interaction and disulfide bonds, and formed soluble aggregates of different sizes. Heating of 5% SPI always promoted large‐size aggregate (LA; >1000 kDa) formation irrespective of temperature, whereas the aggregate size distribution in 2% SPI was temperature dependent: the LA fraction progressively rose with temperature (80→90→100 °C), corresponding to the attenuation of medium‐size aggregates (MA; 670 to 1000 kDa) initially abundant at 80 °C. Heated SPI with abundant LA (>50%) promoted foam stability. LA also exhibited excellent emulsifying activity and stabilized emulsions by promoting the formation of small oil droplets covered with a thick interfacial protein layer. However, despite a similar influence on emulsion stability, MA enhanced foaming capacity but were less capable of stabilizing emulsions than LA. The functionality variation between heated SPI samples is clearly related to the distribution of aggregates that differ in molecular size and surface activity. The findings may encourage further research to develop functional SPI aggregates for various commercial applications.     

Heat stability and freeze–thaw stability of oil-in-water emulsions stabilised by sodium caseinate–maltodextrin conjugates

A sodium caseinate (NaCN)–maltodextrin (Md100) conjugate was prepared by a Maillard-type reaction by dry heat treatment of a NaCN–Md100 mixture at 60 °C and 79% relative humidity for 4 days. Conjugation resulted in a 35.7% loss of available amino groups in the NaCN and a 25.9% loss of available reducing groups in the Md100. The crude conjugate was purified by batch anion exchange chromatography to remove non-conjugated Md100. Purification reduced the available reducing groups in the conjugate from 74.1% to 23.7% and increased the protein content from 45.6% to 83.9%. The emulsifying properties of the conjugates were assessed in oil-in-water (o/w) emulsions; crude and purified conjugate stabilised emulsions had improved storage stability and freeze–thaw stability when compared to NaCN stabilised emulsions. Purified conjugate stabilised emulsions had better thermal stability than NaCN, NaCN–Md mixture and non-purified conjugate stabilised emulsions. These results indicate a potential for these NaCN–Md conjugates as speciality functional food ingredients.     

Effect of limited enzymatic hydrolysis on physico‐chemical properties of soybean protein isolate‐maltodextrin conjugates

The effect of limited hydrolysis was investigated on the physico‐chemical properties of soy protein isolate–maltodextrin (SPI‐Md) conjugate. The hydrolysates at a degree of hydrolysis (DH) of 1.8% showed much higher surface hydrophobicity (H₀; 71.39 ± 3.60) than that of the SPI control (42.09 ± 2.17) and SPI‐Md conjugates (53.46 ± 2.74). Intrinsic fluorescence analysis demonstrated the unfolding of protein molecule and exposure of hydrophobic groups of SPI‐Md conjugate hydrolysates. As evidenced by far‐UV circular dichroism (CD) spectroscopy, the limited hydrolysis increased the unordered secondary structures of SPI‐Md conjugates. The denaturation temperature (T_d) of SPI‐Md conjugate was significantly increased by subsequent limited hydrolysis from 102.53 ± 0.60 °C to 108.11 ± 0.61 °C at DH 1.8%. In particular, the emulsifying activity index (EAI) was improved notably after limited hydrolysis of DH 1.8% (147.76 ± 4.39 m² g^?1) compared with that of native SPI (88.90 ± 1.44 m² g^?1) and SPI‐Md conjugate (108.97 ± 1.45 m² g^?1).     

Gelation enhancement of soy protein isolate by sequential low‐ and ultrahigh‐temperature two‐stage preheating treatments

The effects of combined two heating steps with low (LT, 60 °C for 1 h) and ultrahigh (UHT, 130 or 140 °C for 4 s) temperatures on the thermal gelation of soy protein isolate (SPI) were studied. UHT pretreatments significantly increased protein solubility and enhanced the gelling potential of SPI. Yet, the two‐stage preheating treatment with LT and then UHT‐130 °C had a most remarkable effect: the gel strength of the SPI₆₀₊₁₃₀ sample was, respectively, 1.45‐, 1.64‐ and 3.19‐fold as strong as those of SPI₆₀, SPI₂₅₊₁₃₀, and SPI₂₅. In comparison with single LT or UHT treatments, this two‐stage heating also produced greater amounts of soluble protein aggregates stabilised predominantly by disulphide bonds and hydrophobic forces, contributing to the improved gel network structure.     

Performance of whey protein hydrolysate–maltodextrin conjugates as emulsifiers in model infant formula emulsions

Model infant formula emulsions containing 15.5, 35.0 and 70.0 g L⁻¹ protein, soybean oil and maltodextrin (MD), respectively, were prepared. Emulsions were stabilised by whey protein hydrolysate (WPH) + CITREM (9 g L⁻¹), WPH + lecithin (9 g L⁻¹) or WPH conjugated with MD (WPH–MD). All emulsions had mono-modal oil droplet size distributions post-homogenisation with mean oil droplet diameters (D_4,3) of <1.0 μm. No changes in the D_4,3 were observed after heat treatment (95 °C, 15 min) of the emulsions. Accelerated storage (40 °C, 10 d) of unheated emulsions resulted in an increase in D_4,3 for CITREM (2.86 μm) and lecithin (5.36 μm) containing emulsions. Heated emulsions displayed better stability to accelerated storage with no increase in D_4,3 for CITREM and an increase in D_4,3 for lecithin (2.71 μm) containing emulsions. No increase in D_4,3 over storage was observed for unheated or heated WPH–MD emulsion, indicating its superior stability.     

Comparative studies on the physicochemical properties of soy protein isolate-maltodextrin and soy protein isolate-gum acacia conjugate prepared through Maillard reaction

Dry-heated Maillard reaction was applied in the preparation of protein–polysaccharide conjugates. Reaction mixtures containing soy protein isolate (SPI) and maltodextrin (1:1 weight ratio) were dry-heated at 60 °C and 79% relative humidity for three days. The mixtures of SPI and gum acacia (GA) were dry-heated at the same condition for one week. The conjugate of SPI–MD showed lower levels of free amino groups and higher degree of graft, which indicated that reaction between SPI and MD developed much faster than reaction between SPI and GA. The solubility of SPI at isoelectric point was improved remarkably after grafting with MD or GA. The grafted SPI showed significantly higher levels of emulsifying properties than SPI and the emulsifying properties of SPI–GA conjugate were much better than SPI–MD. Decreases of lysine and arginine contents after the graft reaction indicated that these two amino acid residues attended the covalent linkage between SPI and MD or GA. The graft reaction reduced surface hydrophobicity and fluorescence emission maximum value because of a shielding effect of the polysaccharide chain bound to proteins. The results of secondary structure suggested that grafted SPI had decreased the levels of α-helix, β-sheet and β-turn and increased unordered coils level.     

Formulation of monodisperse oil‐in‐water emulsions loaded with ergocalciferol and cholecalciferol by microchannel emulsification: insights of production characteristics and stability

Nutritional deficiencies of ergocalciferol (VD₂) and cholecalciferol (VD₃) cause skeletal deformations. The primary aim of this study was to encapsulate VD₂ and VD₃ in food‐grade oil‐in‐water (O/W) emulsions by using microchannel emulsification (MCE). Silicon asymmetric straight‐through microchannel (MC) array consisting of 10 313 channels, each having an 11 × 104 μm microslot connected to a 10 μm circular microholes. 1% (w/w) sodium cholate or Tween 20 in water was used as the continuous phase, while 0.5% (w/w) of each VD₂ and VD₃ in different oils served as the dispersed phase. Monodisperse O/W emulsions with Sauter mean diameters of 28 to 32 μm and relative span factor widths below 0.3 were formulated via an asymmetric straight‐through MC array under appropriate operating conditions. The monodisperse O/W emulsions stabilised with Tween 20 remained stable for >30 days with encapsulation efficiencies (EEs) of VD₂ and VD₃ of above 70% at 4 and 25 °C. In contrast, those stabilised with sodium cholate had stability of >30 days with their EEs of over 70% only at 25 °C.     

Cross‐linking of bovine and caprine caseins by microbial transglutaminase and their use as microencapsulating agents for n‐3 fatty acids

Bovine and caprine caseins were cross‐linked with microbial transglutaminase (mTG). The mTG‐cross‐linked bovine or caprine casein dispersion, mixed with 14.5% maltodextrin (DE = 40), was used to prepare emulsions with 10.5% algae oil. Oxidative stability of emulsions was evaluated by peroxide values (PVs) and anisidine values. Adding liposoluble rosemary extract rich in carnosic acid and δ‐tocopherol lowered the formation of hydroperoxides and their subsequent decomposition products in emulsions. Emulsions stabilised with liposoluble rosemary extract rich in carnosic acid and δ‐tocopherol were spray‐dried at 180/95 °C. Algae oil microencapsulated with mTG‐cross‐linked bovine casein reduced PV by ≈ 34%, while the algae oil microencapsulated with mTG‐cross‐linked caprine casein with low levels of α_s1‐casein reduced PV by ≈ 42% at 4 weeks of storage at 30 °C. The investigation suggests that liposoluble rosemary extract rich in carnosic acid and δ‐tocopherol effectively protected algae oil during the coating process with mTG‐cross‐linked bovine and caprine caseins. The above results clearly indicated that the choice of milk caseins (bovine vs. caprine) cross‐linked with mTG impacts the oxidative stability of spray‐dried algae oil emulsions (microcapsules) enriched with n‐3 fatty acids.     

Effect of pre- and post-heat treatments on the physicochemical,microstructural and rheological properties of milk protein concentrate-stabilised oil-in-water emulsions

The effect of heat treatment on the physical stability of milk protein concentrate (MPC) stabilised emulsions was investigated; 3% (w/w) MPC dispersions were preheated at 90 °C for 5 min at neutral pH prior to emulsification. Heat-treated (120 °C, 10 min) emulsions stabilised by preheated MPC had slightly fewer droplet–droplet interactions than that stabilised by unheated MPC because the whey proteins were pre-denatured (∼90% denaturation of the total whey proteins), which led to a reduction in subsequent heat-induced droplet–droplet and droplet–protein interactions. Emulsions stabilised by calcium-depleted MPC were also investigated. The presence of some non-micellar casein fractions gave better emulsification and may have conferred a protective stabilising effect on whey protein aggregation, in both the dispersed phase and the continuous phase during the secondary heat treatment. It was concluded that calcium manipulation and thermal modification of MPC can be utilised to control the functionality in oil-in-water emulsions.     

Formation of acid‐precipitated soy protein–dextran conjugates by Maillard reaction in liquid systems

The conjugation reaction between soybean acid‐precipitated protein (SAPP) and dextran in liquid systems via the initial stage of the Maillard reaction was studied. Functional SAPP–dextran conjugates were prepared in 80% ethanol‐reacting system at 50 °C for 6 h, along with 95% ethanol‐reacting system at 60 °C for 24 h. The covalent attachment of dextran to SAPP was confirmed by sodium dodecyl sulphate–polyacrylamide gel electrophoresis and gel filtration chromatography.Compared to the classical dry‐heating, the reaction time of glycosylation in the two ethanol systems was largely shortened. Emulsifying activity of SAPP–dextran conjugates obtained by dry‐heating incubation and in ethanol was similar at pH 7.0 and10.0, significantly higher than that of SAPP–dextran mixture or SAPP alone. In addition, SAPP–dextran conjugates obtained in 80% ethanol‐reacting system for 6 h were completely soluble after heating at 90 °C for 20 min. The impact of various processing conditions on the formation of SAPP–dextran conjugates was investigated. This study provides important guidance to create protein–polysaccharide conjugates at mild temperatures in liquid systems.     

Impact of whey protein isolates and concentrates on the formation of protein nanoparticles‐stabilised Pickering emulsions

Whey protein nanoparticles (NPs) were prepared by heat‐induced method. The influences of whey protein isolates (WPIs) and concentrates (WPCs) on the formation of NPs were first investigated. Then Pickering emulsions were produced by protein NPs and their properties were evaluated. After heat treatment, WPC NPs showed larger particle size, higher stability against NaCl, lower negative charge and contact angle between air and water. Dispersions of WPC NPs appeared as higher turbidity and viscosity than those of WPI NPs. The interfacial tension of WPC NPs (~7.9 mN/m at 3 wt% NPs) was greatly lower than that of WPI NPs (~12.1 mN/m at 3 wt% NPs). WPC NPs‐stabilised emulsions had smaller particle size and were more homogeneous than WPI NPs‐stabilised emulsions. WPC NPs‐stabilised emulsions had higher stability against NaCl, pH and coalescence during storage.     

Preparation,characterisation and selected functional properties of sodium caseinate–maltodextrin conjugates

Sodium caseinate (NaCN)–maltodextrin (Md40 or Md100) conjugates were prepared by a Maillard-type reaction by dry heat treatment of mixtures of NaCN and Md at 60 °C and 79% relative humidity for 4 days. Minimal levels of coloured reaction products were formed during conjugate preparation. Conjugation resulted in a 35.6% and a 36.2% loss of available amino groups in the NaCN, and a 17.8% and a 25.7% loss of available reducing groups in Md40 and Md100, respectively. SDS–PAGE and gel permeation chromatography confirmed conjugation. When assessed in the pH range 2.0–8.0 at 20 °C and 50 °C, conjugates had improved solubility compared to NaCN, particularly around the isoelectric point of the protein. The emulsifying properties of NaCN–Md conjugates were assessed in oil-in-water (o/w) emulsions and in model cream liqueurs. The conjugate stabilised o/w emulsions and liqueurs showed improved stability when compared to NaCN stabilised o/w emulsions and liqueurs. These results indicate a potential for these NaCN–Md conjugates as speciality functional food ingredients.     

Emulsifying Properties of Legume Proteins Compared to β‐Lactoglobulin and Tween 20 and the Volatile Release from Oil‐in‐Water Emulsions

The emulsifying properties of plant legume protein isolates (soy, pea, and lupin) were compared to a milk whey protein, β‐lactoglobulin (β‐lg), and a nonionic surfactant (Tween 20). The protein fractional composition was characterized using sodium dodecyl sulfate–polyacrylamide gel electrophoresis analysis. The following emulsion properties were measured: particle diameter, shear surface ζ‐potential, interfacial tension (IT), and creaming velocity. The effect of protein preheat treatment (90 °C for 10 min) on the emulsifying behavior and the release of selected volatile organic compounds (VOCs) from emulsions under oral conditions was also investigated in real time using proton transfer reaction‐mass spectrometry. The legume proteins showed comparable results to β‐lg and Tween 20, forming stable, negatively charged emulsions with particle diameter d_3,2 < 0.4 μm, and maintained stability over 50 d. The relatively lower stability of lupin emulsions was significantly correlated with the low protein surface hydrophobicity and IT of the emulsion. After heating the proteins, the droplet size of pea and lupin emulsions decreased. The VOC release profile was similar between the protein‐stabilized emulsions, and greater retention was observed for Tween 20‐stabilized emulsions. This study demonstrates the potential application of legume proteins as alternative emulsifiers to milk proteins in emulsion products.     

Emulsifying properties of proteins extracted from Australian canola meal

Canola protein albumin fraction, globulin fraction, and canola protein isolate (CPI) were compared to commercial soy protein isolate (SPI) in terms of their emulsifying properties at various pH values. The globulin fraction had higher emulsifying capacity (EC), higher emulsifying activity index (EAI), and the droplet size of emulsions it stabilized was consistently smaller irrespective of pH compared to albumin fraction or CPI. In comparison to SPI, globulin fractions also had higher EC at all pH values tested, higher EAI at acidic pH, and smaller or comparable average emulsion droplet size at both pH 4 and 7. The stability of canola protein based emulsions were comparable to those of SPI based emulsions at most pH values (except the emulsion stabilized by the CPI at pH 4), with no significant (p > 0.05) changes in droplet size during storage for up to 7 days at room temperature. These emulsions, however, experienced separation into the emulsion and serum phases after 24 h storage at room temperature with the exception of CPI- and SPI-stabilized emulsions at pH 9. This study demonstrates the comparable emulsifying properties (forming or stabilizing) of some canola proteins to commercially available SPI, suggesting the potential use of canola proteins in food applications.     

Enhancement of the functional properties of whey protein by conjugation with maltodextrin under dry‐heating conditions

Conjugation of whey protein isolate (WPI) and maltodextrin (MD, dextrose equivalent of 6) was achieved by dry‐heating at an initial pH of 7.0, at 60 °C and 79% relative humidity, with WPI: MD6 ratio of 1:1, for up to 24 h. Conjugation was achieved with limited development of colour and advanced Maillard products on 24 h of heating. Conjugation increased the protein solubility at pH 4.5, by 7.1–8.5%, compared to the unheated and heated WPI controls. Conjugation of WPI with MD6 enhanced the stability and retention of clarity in protein solutions heated at 85 °C for 10 min with 50 mM added NaCl.     

Physicochemical stability of egg protein‐stabilised oil‐in‐water emulsions supplemented with vegetable powders

The effect of vegetable powders on the physicochemical stability of egg protein‐stabilised oil‐in‐water emulsions was studied. Vegetable powders (beetroot, broccoli, carrot, celery, green pea, red pepper, spinach, swede, tomato and yellow pea) were added at 2.5% (w/v) to emulsions prepared with rapeseed oil. The physical stability of the emulsions was characterised using the emulsifying activity (EAI) and the emulsifying stability indices (ESI) in addition to bright field microscopy. The oxidative stability of the emulsions was monitored by means of an accelerated oxidation test (Rancimat method). The addition of most vegetable powders did not markedly affect the physical stability of the emulsions although an adverse effect of tomato was observed. The oxidative stability of the emulsions was significantly improved in most cases as indicated by the Rancimat method with broccoli exhibiting the highest increase in induction time (98.2%) compared with the control. Both polar and nonpolar antioxidants are likely to contribute to the overall chemical stability of this complex food system in a concentration‐dependent manner.