Edible mayonnaise-like emulsion stabilized by rice bran protein fibril aggregation: Effect of fibril aggregate structure

Edible mayonnaise-like emulsion stabilized by rice bran protein fibril aggregation: Effect of fibril aggregate structure | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Article Edible mayonnaise-like emulsion stabilized by rice bran protein fibril aggregation: Effect of fibril aggregate structure Junqi Pang, Yuguang Zhang, Keyang Sun, Wanyue Jiang, Xuesi Pan, and 2 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-7678095/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 12 Jan, 2026 Read the published version in npj Science of Food → Version 1 posted 13 You are reading this latest preprint version Abstract It was investigated the impact of rice bran protein (RBP) fiber (PF) aggregates on the properties of mayonnaise-like emulsions. Using RBP with different structures during PF aggregate formation, plant-based emulsions were prepared and evaluated for interfacial protein adsorption (AP%), water-holding capacity (WHC), rheological and textural properties, color, sensory quality, and storage stability. PF aggregate–based emulsions had 11.29% and 16.8% higher AP% values and 43.73% and 107.64% higher WHC values than those of RBP emulsions and homemade mayonnaise, respectively. Texture analysis showed significant improvements in hardness, viscosity, cohesiveness, and springiness. After 30 days, peroxide values in PF aggregate emulsions were 25.13% lower than those in RBP emulsions and 33.57% lower than those in homemade mayonnaise; malondialdehyde content was 15.57% and 21.25% lower, respectively. The total viable count of PF aggregate emulsions was 21.51% lower than that of RBP emulsions. These findings highlight the enhanced stability of PF aggregate–based mayonnaise-like emulsions. Physical sciences/Chemistry Physical sciences/Engineering Physical sciences/Materials science Rice Bran Protein Protein Fiber Aggregates Mayonnaise-like Products Rheological Properties Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 1. Introduction Mayonnaise is a widely used condiment in many dishes. However, its health and application drawbacks have recently attracted consumer attention 1 . The main concerns include the following: first, its high caloric and fat content, as mayonnaise is primarily composed of oil and egg yolks, which can contribute to obesity and related health issues 2 ; second, its cholesterol and sodium levels, which, with long-term excessive intake, may affect blood pressure and cardiovascular health and are unsuitable for individuals with high cholesterol 3 ; and third, the presence of egg components, which makes it inappropriate for individuals with egg allergies 4 . Because of these limitations, some consumers prefer healthier alternatives or low-fat versions to better align with their dietary and health needs 5 . Consequently, innovating formulations to create healthier and more diverse mayonnaise products has become a significant goal for industry development 6 . Plant-based mayonnaise products have gained popularity due to their environmental, health, and sustainability benefits. Compared with traditional animal-based mayonnaise, their production generally requires less carbon and water, making them more environmentally friendly 7 . Moreover, plant proteins are cholesterol-free and rich in unsaturated fatty acids, making them suitable for formulating low-cholesterol mayonnaise. From a sourcing perspective, plant proteins are more widely available and can contribute to agricultural development 8 . Commonly used plant proteins in mayonnaise production include pea, soy, and chickpea proteins. However, not all plant proteins are suitable for mayonnaise-like products, as they must exhibit strong emulsifying and textural properties to replicate the texture and mouthfeel of traditional mayonnaise. To address this, the modification of plant proteins to improve their emulsifying functionality is a common industrial strategy 9 . Rice bran protein (RBP), extracted from rice bran (a byproduct of rice milling with a protein content of 10%–16%), is typically obtained using an alkali-soluble acid-precipitation method from defatted rice bran. Its unique amino acid composition imparts nutritional value and potential for industrial applications 10 . RBP is a low-allergenic, high-quality plant protein with desirable functional properties such as solubility, emulsification, and foaming 11 . However, its native structure limits its ability to meet diverse market demands, necessitating modifications to enhance its functionality 12 , 13 . In recent years, various physical, chemical, and enzymatic modification strategies have been widely employed for this purpose 14 . Among these, acid-thermal treatment of RBP to generate fibril structures has proven particularly effective and desirable for practical applications. The resulting fibrils, characterized by abundant cross-β-sheets and “zipper-like” configurations, provide excellent stability under extreme conditions. This treatment improves multiple functional properties of RBP, including viscosity, gelation, emulsification, foaming, antioxidant activity, and antimicrobial activity, highlighting its significant potential for future applications in food processing 15 . Protein modification to form fiber aggregates enhances the functional properties of proteins. Compared with native proteins, fiber aggregates show significant improvements in gelation, emulsifying activity, and foaming properties 16 . The superior gelation capacity of protein fiber (PF) aggregates results from their ability to cross-link with polysaccharides in aqueous environments, forming hydrogels with strong water retention and elasticity. These properties enable their use in food applications as thickeners to achieve desired consistency and texture 17 . PF aggregates also exhibit enhanced emulsifying and foaming performance. Owing to their higher aspect ratio, fiber structures demonstrate superior emulsifying effects and are widely used in the preparation of high internal phase emulsions 18 . Studies have further shown that PFs can stabilize Pickering emulsions through irreversible interfacial adsorption and anti-aggregation properties while improving the bioavailability of encapsulated nutrients. Additionally, surface hydrophobicity has been identified as a key factor influencing the emulsifying and foaming behavior of fiber aggregates 19 . Importantly, the quality and stability of plant-based mayonnaise products are directly linked to the emulsifying and gelation properties of the proteins employed 20 . Thus, developing mayonnaise-like products using plant PF aggregates holds significant potential. Herein, RBP aggregates with different structures, prepared by heating them for varying durations at pH 2.0, were used to develop mayonnaise-like products. The properties of these products, including rheology, color differences, interfacial protein adsorption (AP%), texture, water-holding capacity (WHC), sensory attributes, and storage stability, were evaluated to assess the performance of mayonnaise-like products formulated with RBP fiber structures. The results were then compared with those of traditional egg yolk–based mayonnaise to verify the feasibility and practicality of using fiber-structured RBP in the production of mayonnaise-like products. 2. Methods 2.1 Materials Defatted rice bran was provided by Shenyang Aoda Oil (Liaoning, China).Fresh eggs were purchased from the local market (Shenyang, China). Soybean oil was provided by Yili Goli Company (Shanghai, China). Salt was provided by Yunnan Salt Industry Co., Ltd (Yunnan, China). White sugar and White vinegar were provided by Lianhua Health Industry Group Co., Ltd (Henan, China). Anhydrous ethanol, sodium hydroxide and hydrochloric acid were provided by Tianjin Hengxing Chemical Reagent Manufacturing Co., Ltd. (Tianjin, China). Isooctane, Isopropanol, N-butanol, Trichloroacetic acid, Thiobarbituric acid, Potassium thiocyanate, Ferrous sulfate, Barium chloride, 1,1,3,3-tetraethoxypropane and Normal saline all purchased from Shanghai Aladdin Biochemical Technology Co., Ltd. (Shanghai, China), all the purities are at the analytical purity level. All aqueous solutions were prepared using distilled water. 2.2 Experimental Method 2.2.1 Protein extraction A previously described method 21 was followed with slight modifications. Defatted rice bran was mixed with 10 volumes of distilled water. The pH was adjusted to 9.5 with 1.0 mol/L NaOH. The mixture was stirred at 45 ℃ for 2 h, and then centrifuged at a speed of 4000 r/min for 15 min using the centrifuge (LD4-2, Beckmann Kurtz Trading Company, USA). The supernatant was collected and adjusted to pH 4.5 with 1.0 mol/L HCl, followed by centrifugation at 4000 r/min for 15 min. The precipitate was adjusted to a neutral pH. The mixture was then freeze-dried using the freeze-drying machine (SCIENTZ-12NIA, Ningbo Xinzi Biotechnology Co., Ltd., China). The protein content of the freeze-dried powder was determined using the Kjeldahl method. 2.2.2 Preparation of protein fiber polymers A 10 mg/mL RBP solution was prepared and stirred at 40°C for 2 h to ensure complete dissolution. The pH was adjusted to 2.0, and the solution was then heated at 90°C for varying durations (0, 2, 4, 6, 8, 10, and 12 h). After heating, the samples were immediately cooled in an ice-water bath to obtain protein solutions at several stages of the PF preparation process. 2.2.3 Atomic Force Microscopy (AFM) images The RBP samples were diluted to a concentration of 0.02 mg/mL with distilled water (pH 2.0). Subsequently, 5 µL of the diluted sample was deposited onto a mica surface and dried at 25°C for 24 h. AFM images were taken using a Dimension Icon AFM with an SNL-10 cantilever probe in tapping mode (Bruker Corporation, Billerica, MA, USA). AFM images were analyzed with NanoScope Analysis version 1.90 and FiberApp software. The dimensions of the fibrils were measured using the cross-sectional tool in NanoScope Analysis version 1.90. FiberApp software, which is designed to track and analyze the contour lengths of flexible and rigid fibrils, was used to make the calculations. 2.2.4 Preparation of PFs stabilized Mayonnaise-like emulsions The traditional mayonnaise preparation was prepared as follows 5.0 mL of egg yolk was weighed into a clean 50-mL centrifuge tube. 0.2 g salt, 0.4 g white sugar, and 0.5 g white vinegar were added. The mixture was stirred until the salt and sugar were completely dissolved. Subsequently, 15.0 mL soybean oil was added to the centrifuge tube, and the mixture was homogenized at 16000 r/min for 2 min. PF-stabilized mayonnaise was prepared as follows: 5.0 mL PF was prepared by heating at pH 2.0 (10 mg/mL) and was weighed into a clean 50-mL centrifuge tube. 0.2 g salt, 0.4 g white sugar, and 0.5 g white vinegar were added, and the mixture was thoroughly stirred until the salt and sugar were dissolved. Subsequently, 15.0 g soybean oil was added to the centrifuge tube. The mixture was homogenized at 16000 r/min for 2 min. 2.2.5 Protein adsorption (AP%) The AP% was measured using a previously described method 22 . A 20 mL aliquot of the freshly prepared mayonnaise-like product was centrifuged at 4000 r/min for 10 min. The lower transparent layer of the separated liquid was gently aspirated. Its protein concentration was determined using the Coomassie Brilliant Blue method. The AP% of the mayonnaise-like product was then calculated as follows: $$\:AP\%=\frac{{C}_{S}-{C}_{f}}{{C}_{s}}\times\:100$$ In the formula, C s represents the total protein content in the mayonnaise-like product (mg/mL), and C f indicates the protein content in the aqueous phase after centrifugation (mg/mL). 2.2.6 Water Holding Capacity (WHC) The prepared mayonnaise-like emulsions was transferred into a 50mL centrifuge tube and weighed to determine the initial mass (m 1 ). The sample was then centrifuged at 6000 r/min for 5 min. The water separated in the centrifuge tube was removed, and the tube was weighed again to find the final mass (m 2 ). The WHC of the mayonnaise-like product was calculated using the following formula: \(\:\text{W}\text{H}\text{C}（\text{%}）=\frac{{\text{m}}_{2}-{\text{m}}_{0}}{{\text{m}}_{1}-{\text{m}}_{0}}\times\:\) 100 23 In the formula, m 0 is the mass of the centrifuge tube, m 1 is the total mass of the centrifuge tube and egg yolk sauce before centrifugation, and m 2 represents the total mass of the centrifuge tube and egg yolk sauce after centrifugation. 2.2.7 Rheological properties The rheological properties of the mayonnaise-like emulsions were immediately examined after preparation with a hybrid rheometer (DHR-1, 10 nN·m-50 mN·m; TA Instruments, New Castle, DE, USA) to measure both dynamic and steady-state shear rheological properties. Measurements were taken using parallel plate geometry with a diameter of 40 mm and a gap of 1000 µm. First, the mayonnaise-like product viscosity was assessed by flow sweep mode at shear rates ranging from 0.01 to 100 s − 1 at a constant temperature of 25°C, and the viscosity ratio was calculated over a range of 0.1–100 s. For the dynamic oscillation scanning tests, measurements were also taken with a parallel plate with a diameter of 40 mm and a gap of 1000 µm. The tested emulsion was placed on the platform, and the plate was pressed down to the trim distance. Excess sample around the plate was removed. The sample was sealed with silicone oil. The storage modulus (G') and loss modulus (G'') of the mayonnaise-like product were determined at a strain value of 0.2% and a temperature of 25°C using the angular frequency sweep mode. 2.2.8 Texture properties The texture properties of the mayonnaise-like emulsions were evaluated using an established method 24 , with slight modifications. The samples were prepared in 50mL centrifuge tubes and subjected to texture analysis at 25°C with a TA-XT Plus texture analyzer equipped with a P/36R cylindrical probe. Hardness, adhesiveness, cohesiveness, springiness, and chewiness were assessed. The measurement parameters were set as follows: the pre-test speed was 1.0 mm/s, the test speed was 0.6 mm/s, the post-test speed was 1.0 mm/s, the deformation degree was 30%, and the trigger force was 0.3 g. Each sample was tested in triplicate, and the average values are reported. 2.2.9 Color difference 10 mL of the freshly prepared mayonnaise and mayonnaise-like emulsions were placed into display bottles. The colorimeter was calibrated using a white calibration plate. The L* (lightness), a* (redness), and b* (yellowness) values were measured for each sample. Each sample was measured in triplicate. The average values were recorded. 2.2.10 Sensory evaluation Sensory evaluation of the mayonnaise products was performed by a semi-trained panel of 10 food science students. The panel assessed the color, consistency, aroma, flavor, and overall acceptability of the samples. The average score assigned by the panelists for each attribute served as the evaluation score for that attribute. The scoring criteria were as follows: each attribute could be given a maximum of 20 points. Scores ranging from 17 to 20 indicated high satisfaction with the product, whereas scores between 12 and 16 indicated moderate satisfaction, and scores below 12 indicated dissatisfaction with the product quality. 2.2.11 Storage Quality Properties of the Mayonnaise-like emulsions at Room Temperature The storage stability of mayonnaise and the mayonnaise-like product was evaluated using a room-temperature test. Freshly prepared mayonnaise and the mayonnaise-like product were stored at room temperature for 30 d. Measurements were taken every 3 d, with three samples drawn each time and testing performed in triplicate. Indicators tested included peroxide value, malondialdehyde (MDA) content, and total bacterial count. 2.2.11.1 Peroxide Value The peroxide value determination in mayonnaise was based on the standard GB 5009.227–2023. The specific procedure was as follows: 50 µL freshly prepared mayonnaise and mayonnaise-like product prepared with PF were accurately dispensed into a clean 5mL centrifuge tube. 3 mL heated iso-octane/isopropanol (3:1, V/V) was added, and the samples were thoroughly mixed using a vortex mixer. The mixture was centrifuged at 8000 r/min for 5 min. 0.2 mL of the supernatant was mixed with 2.8 mL methanol/n-butanol (2:1, V/V). 0.15 mL freshly prepared FeCl 2 solution (0.25 mol/L) and 0.15 mL potassium thiocyanate solution (0.3 g/mL) were added. The mixture was allowed to react in the dark for 20 min. Measure the absorbance using an ultraviolet spectrophotometer (UV-1200S, Aowei Instrument, China) and the absorbance was measured at 510 nm. The measured absorbance values were used to calculate the peroxide value of the mayonnaise by referencing the standard curve established with Fe 3+ solutions. The standard curve was established using Fe 3+ solutions with concentrations of 0.02, 0.04, 0.08, 0.16, and 0.20 mmol/L using a methanol:n-butanol (2:1, V/V) mixture as the blank. The equation of the standard curve is y = 4.50312x − 0.0081 (R² = 0.9905). Each sample was measured in triplicate, and the average value was calculated. 2.2.11.2 MDA content MDA content in mayonnaise was measured in accordance with GB 5009.181–2016. The standard curve was generated using 1,1,3,3-tetraethoxypropane, according to the equation y = 1.04561x + 0.0035 (R 2 = 0.9991). 2.2.11.3 Total viable count The determination of the total viable count (TVC) in mayonnaise during storage was performed in accordance with GB 4789.2–2022, the National Food Safety Standard for Microbiological Examination of Food, specifically the method for total viable count determination. 2.2.12 Statistical analysis All measurement results were tested three times, from which the average value and standard deviation value were calculated. The statistical differences, mean values and standard deviations of the data were analyzed using the SPSS 24.0 software package. Furthermore, multiple comparisons were conducted using the Tukey test. Different letters indicate p < 0.05. Origin 2022 is used to complete the drawings. 3. Results and Discussion 3.1 AFM images of PFs AFM is a high-resolution microscopic technique enabling direct observation of the microscopic morphology of RBP fiber aggregates 25 . AFM images were used to study the microscopic morphological changes of RBP heated for 0, 6, and 12 h at pH 2.0. The results are shown in Fig. 1 . At pH 2.0, PFs formed after heating for 0 h presented large spherical particles with a chain width of approximately 300 nm and a chain height of approximately 23 nm. The overly large particle structure was not suitable as an emulsifier at the oil-water interface. After 12 h of heating at pH 2.0, PFs formed, and multiple PFs with slender fiber structures could be seen in the field of view. Under other heating times, these structures were not clearly visible. Compared to the PFs formed after heating for 6 h, the chain width and chain height of these PFs increased by 25.0% and 36.5%, respectively. The larger chain width and chain height given the mayonnaise-like emulsions stabilized from PFs better stability 26 . This result indicated that chose the appropriate heating time was crucial for the formation of RBP fibers. The heating time changed the secondary structure and surface charge state of the protein, thereby affected the intermolecular interactions and promoting the formation of fibers 27 . 3.2 AP% analysis In emulsion systems, the level of AP% has a significant impact on the stability and functional properties of the emulsion 28 . The AP% of the mayonnaise-like emulsion made with RBP and the mayonnaise prepared with egg yolk are shown in Fig. 2 -a. The homemade mayonnaise made with egg yolk exhibited the poorest performance in terms of AP%. This is attributed to its larger droplet size and less dense distribution of the emulsion droplets in mayonnaise prepared by egg yolk homogenization. Conversely, the mayonnaise-like emulsion prepared with RBP aggregates heated for different times at pH 2.0 showed no significant difference in AP%. Only the mayonnaise-like emulsion prepared with the native protein without heating exhibited significantly lower AP% because the hydrophobic groups in the native protein are protected within the protein matrix and are not exposed 29 . After heating for 12 h, RBP forms elongated fibrous structures. This process modifies the native RBP, altering its surface structure and chemical properties, thereby increasing its adsorption at the oil–water interface in the mayonnaise-like emulsion 30 . Changes in the surface charge distribution of the protein accompany the formation of fibrous structures. If the surface carries more charge that can interact with the oil–water interface, its adsorption capacity is enhanced 31 . Additionally, studies have shown that protein fibrous structure formation reduces the molecular weight of the protein. Smaller molecular weight fragments may more easily diffuse to and adsorb at the oil–water interface 32 . 3.3 WHC analysis WHC is one of the fundamental characteristics for evaluating the structural stability of emulsions. It reflects the ability of the emulsion system to retain water 33 . The WHC of various mayonnaise samples is shown in Fig. 2 -b. The mayonnaise prepared with egg yolk showed the poorest WHC. This is because the aqueous phase is relatively abundant during mayonnaise preparation, and the proteins and lecithin in the egg yolk alone cannot stabilize the mayonnaise emulsion. Among the mayonnaise-like emulsions prepared with RBP, the one with the highest WHC is the emulsion stabilized by PF obtained after heating for 12 h at pH 2.0, because the fibrillar proteins possess a highly ordered β-sheet structure. This tightly packed fibrous morphology provides a large surface area and a multipoint hydrogen-bonding network, facilitating water capture and retention. Additionally, the fibers form larger aggregates, which can adsorb more water, enhancing WHC 34 . In contrast, the native structure of RBP restricts its interaction with water molecules, resulting in a poorer WHC. 3.4 Rheological properties analysis The droplet aggregation state in mayonnaise-like emulsions determines their rheological properties. As shown in Fig. 4 , not all mayonnaise-like emulsions stabilized with PFs are supported viscoelastic emulsions that can maintain sharp edges. Only the PFs stabilized mayonnaise-like emulsions formed after heating at pH 2.0 and 90 ℃ for 8 h showed a viscoelastic emulsion with a supporting structure. Moreover, the longer the heating time, the better the edge retention. As shown in Fig. 3 -a, all the mayonnaise-like emulsions samples exhibited G' >G", indicating that the emulsion was mainly elastic 35 . In the RBP heated to form PFs stabilized mayonnaise-like emulsions at pH 2.0, the sample group heated for 12 h exhibited the best rheological properties, with G' and G" higher than those of other samples. Combined with AFM image analysis, this is because the PFs can form a network structure, better adsorbing on the oil-water interface, making the mayonnaise-like emulsions exhibit more stable properties 25 . Therefore, acid heating modification of RBP to obtain a performance suitable for industrial production of egg yolk sauce analogues has significant commercial value. As the scanning frequency increased, the G' and G" values of all sample groups except the PFs-0 h stabilized mayonnaise-like emulsions remain relatively stable. This once again indicates that the large particles of protein, as emulsifiers, have poor stability. When affected by external forces, they are prone to undergo undesirable changes in properties, which is not conducive to industrial production. The frequency-independent rheological behavior of PFs prepared under pH 2.0 conditions can be attributed to their high protein adsorption on the emulsion interfacial membrane, which enhanced the interaction between droplets. The curves showed the viscosity changes of mayonnaise and PFs stabilized mayonnaise-like emulsions with respect to shear rate are shown in Fig. 3 -b. All samples exhibited shear thinning behavior, which was explained by the coagulation of emulsion droplets 36 . However, among these samples, the mayonnaise-like emulsions of PFs-12 h that was stable showed the highest viscosity at the beginning of shear. This was due to the fibrous structure of PFs-12 h, which allowed more protein adsorption at the emulsion interface. As the heating time increased, the RBP structure unfolded and exposed hydrophobic groups, thereby promoting the formation of smaller droplets during the emulsion preparation process and causing a more compact droplet arrangement 37 . 3.5 Texture properties analysis The textural properties of mayonnaise are an essential factor influencing its sensory characteristics and consumer acceptance. Texture profile analysis, conducted using a texture analyzer, involves two compressions of the samples to measure parameters such as hardness, adhesiveness, springiness, and cohesiveness. The textural properties of the mayonnaise-like emulsions prepared from RBP aggregates and the homemade mayonnaise are shown in Table 1. Table.1 Texture properties of different PFs mayonnaise-like and mayonnaise Condition Heating time/h Hardness/N Viscosity/mj Cohesion Elasticity/mm pH = 2.0 0 3.2 ± 0.2 i 0.61 ± 0.05 j 0.65 ± 0.04 hi 0.72 ± 0.03 ij 2 3.45 ± 0.09 hi 0.83 ± 0.05 gh 0.75 ± 0.03 fg 0.85 ± 0.04 hi 4 3.79 ± 0.03 gh 0.93 ± 0.03 g 0.81 ± 0.02 f 1 ± 0.04 g 6 4.38 ± 0.19 ef 1.08 ± 0.03 f 0.9 ± 0.04 e 1.29 ± 0.12 f 8 5.28 ± 0.18 d 1.39 ± 0.05 e 0.97 ± 0.03 de 1.64 ± 0.06 e 10 5.47 ± 0.27 d 1.63 ± 0.04 d 1.05 ± 0.03 d 1.82 ± 0.05 d 12 6.28 ± 0.22 c 1.9 ± 0.03 c 1.3 ± 0.07 c 2.04 ± 0.06 c egg yolk 7.65 ± 0.22 b 3.5 ± 0.27 d 1.81 ± 0.04 b 8.32 ± 0.18 b Note: Different small letters in the same column indicate a significant difference ( p < 0.05). Mayonnaise prepared from egg yolk demonstrated superior textural properties, attributed to the presence of emulsifying components in egg yolk. For the RBP-based mayonnaise-like emulsions, different heating durations at pH 2.0 significantly affected their textural properties. Specifically, the overall textural performance of samples prepared at pH 2.0 was higher than that of mayonnaise prepared with egg yolk. This is because the pH environment affects the protein structure in the solution, causing denaturation, thereby influencing the emulsifying properties of the proteins. Under pH 2.0, the longer the RBP solution was heated, the better the hardness and viscosity of the resulting mayonnaise-like emulsion, and the higher its cohesiveness and springiness 23 . This trend is consistent with the changes in elasticity and viscosity observed in the rheological measurements. 3.6 Color differences analysis The color differences of mayonnaise-like emulsions stabilized from RBP heated under pH 2.0 conditions and other mayonnaise samples are shown in Fig. 4 . The L* value, which describes the brightness of a color, indicates how light or dark an object is. The a* value represents how red or green it is, with positive values indicating higher redness and negative values indicating higher greenness. The b* value represents whether the color is more yellow or blue, with positive values indicating higher yellowness and negative values indicating higher blueness. For mayonnaise-like emulsions prepared with RBP, the L* values were all > 80, indicating that the emulsions were very bright and close to white. The a* values of all samples were very low; manufacturers typically use this value to help distinguish differences in hue between batches or brands. All mayonnaise samples had relatively high b* values, especially those prepared from egg yolk, due to the inherent color of the egg yolk itself. For RBP-based mayonnaise-like emulsions, there was significant variation in the b* values of emulsions prepared under pH 2.0 conditions. This is because the pH of the protein solution has a substantial impact on the RBP color, which in turn affects the color of the mayonnaise-like emulsion. Specifically, the color of the protein solution becomes darker or lighter at different pH levels, possibly due to protein denaturation or changes in specific amino acid residues. RBP is composed of various amino acids, whose side chains can ionize at different pH levels. For example, glutamic acid and aspartic acid protonate at low pH and deprotonate at high pH. In terms of color acceptability, the mayonnaise-like emulsion prepared from RBP after heating for 12 h at pH 2.0 was more generally accepted. This is particularly important for developing healthier mayonnaise products. 3.7 Sensory evaluation analysis The sensory evaluation of mayonnaise-like emulsions prepared from RBP aggregates and homemade mayonnaise is shown in Fig. 5 . The sensory scores of the RBP-based mayonnaise-like emulsions were typically higher than those of homemade mayonnaise prepared with egg yolk. For the egg yolk mayonnaise, the lack of additives and the yolk’s inherent color and odor contributed to its lower sensory scores. Conversely, for RBP-based mayonnaise-like emulsions, those prepared after heating RBP for 12 h at pH 2.0 received the highest sensory scores for viscosity, because the emulsification process requires homogenization, which involves dispersing oil into fine droplets uniformly distributed in the aqueous phase. Heating RBP for 12 h at pH 2.0 results in the formation of elongated fibrous structures, contributing to this homogenization process. 3.8 Determination of Quality and Performance of Mayonnaise Stored at Room Temperature 3.8.1 Peroxide values analysis The oxidation of soybean oil in mayonnaise and mayonnaise-like emulsions is a complex process that produces both primary and secondary oxidation products. During the initial oxidation stage, the unsaturated fatty acids in soybean oil—such as linoleic acid and linolenic acid—react with oxygen to form peroxides. We measured the oxidation level of mayonnaise and mayonnaise-like emulsions by assessing the primary oxidation product, hydrogen peroxide. Figure 6 -a illustrates changes in the hydrogen peroxide content in mayonnaise-like emulsions prepared with RBP over 30 days of storage. As shown in the figure, both freshly prepared egg yolk mayonnaise and mayonnaise-like emulsion samples exhibited high peroxide values (1.54 mmol/kg). This may be due to the high-speed homogenization (16,000 r/min) used in the emulsion preparation process, which increases the emulsion system’s temperature and enhances the contact between oxygen and soybean oil, leading to the formation of oxidative products. Figure 6 -a demonstrates that among the mayonnaise-like emulsions, the samples prepared with RBP after 12 h of heating had the lowest peroxide values. This is attributed to the high structural stability, strong antioxidant properties, dense network protection, and effective interfacial physicochemical characteristics of the fibrillar protein structure 38 . Therefore, using fibrillar RBP significantly enhances the mayonnaise-like emulsion’s stability during storage, extending the product’s shelf life, and inhibiting the oxidation of oils. As shown in Fig. 6 -a, the peroxide values of mayonnaise-like emulsions made with RBP were significantly lower than those of mayonnaise prepared with egg yolk. The oxidation rate of soybean oil in mayonnaise-like emulsions is affected by several factors: fibrillar protein structure formation in RBP under acidic conditions; the dense droplet structure of mayonnaise-like emulsions prepared at pH 2.0, which reduces the contact between droplets and oxygen, lowering the oxidation rate; and the enhanced activity of certain antioxidants under acidic conditions, which contributes to the antioxidant properties of mayonnaise-like emulsions. 3.8.2 MDA content analysis Soybean oil in mayonnaise oxidizes when exposed to oxygen, producing peroxides. The major oxidation products are mainly hydroperoxides and other primary peroxides, which break down into free radicals under the influence of heat, light, or metal ions, leading to further oxidation. These free radicals react with nearby molecules to form secondary oxidation products like aldehydes (e.g., MDA), carboxylic acids, and ketones, which are typically the sources of negatively perceived flavors. Figure 6 -b shows the relationship between MDA production and storage time over 30 d. As shown in the figure, the rate of increase in MDA production was rapid during the first 15 d of storage but slowed over time. This is because MDA is a secondary product formed from primary oxidation products. As the oxidation process continues, the consumption of oxygen slows the primary oxidation process, reducing the primary metabolite hydroperoxide consumption, consequently decreasing secondary oxidation rate. As shown in Fig. 6 -b, both RBP-based mayonnaise-like emulsions and homemade mayonnaise exhibited a similar trend in MDA production during storage. However, there were significant differences in the quantities produced. The mayonnaise-like emulsion stabilized by RBP heated for 12 h at pH 2.0 had a MDA content of 1.22 mg/kg, which was significantly lower than that of the other RBP-based emulsions. This trend is consistent with the previously observed changes in peroxide values. This can be attributed to the greater stability of its fibrous RBP structure, resulting in fewer hydroperoxides being generated during storage compared to other samples, reducing the rate of secondary oxidation and leading to lower MDA production. 3.8.3 Changes in Total Bacterial Count analysis TVC determination is a key method for indicating the microbial contamination level in food. A high TVC can indicate the presence of pathogenic bacteria, which poses a threat to consumer health. Changes in the TVC of mayonnaise-like emulsions prepared with heated RBP at pH 2.0 over 30 days of storage are shown in Fig. 6 -c. Since vinegar was added during mayonnaise preparation, the acetic acid in vinegar lowered the pH of the emulsion system. A low-pH environment is conducive to bacterial growth inhibition. Based on this, the TVC of RBP-based mayonnaise-like emulsions prepared at pH 2.0 should increase less during storage compared to homemade mayonnaise. As shown in Fig. 6 -c, the measured results are consistent with this prediction. Even after 30 days, the TVC of the mayonnaise-like emulsion remained < 900 CFU/ml. Therefore, RBP-based mayonnaise-like emulsions prepared under pH 2.0 conditions do not require preservatives in industrial production. Moreover, the TVC of mayonnaise-like emulsions prepared from fibrillar RBP after 12 h of heating at pH 2.0 is even lower, indicating that this type of emulsion has great potential for developing mayonnaise products. Conclusions The mayonnaise-like emulsion stabilized by PFs exhibited superior rheological properties with both G′ and G′′ values higher than those observed under other treatment conditions. This significantly enhanced its shear-thinning behavior and viscoelasticity, as reflected in the high hardness, viscosity, and cohesiveness measured in the textural analysis. The RBP-based emulsions also showed improved color, and in sensory evaluation, the pH 2.0–treated samples achieved higher scores for consistency and acceptability, attributable to the formation of fibrous structures. These findings are of practical significance for developing healthy, additive-free mayonnaise-like products. Furthermore, the PF-stabilized emulsion prepared under these conditions demonstrated lower MDA production and peroxide values during storage, indicating strong resistance to lipid oxidation. Thus, PF formation contributes to both the stability and desirable characteristics of mayonnaise-like products. Overall, this study provides a valuable reference for the development of plant protein–based mayonnaise alternatives. Declarations Data availability All data generated or analyzed during this study are included in this published article. Received: XXX; Accepted: XXX; Published online:XXX Acknowledgments This research was supported by Liaoning Province Applied Basic Research Program, (1746603820076); 2025 Liaoning Provincial Department of Education Platform Construction Project (LJ232510166005); Special Plan for Science and Technology Special Envoy Action of Liaoning Provincial Department of Science and Technology (1741935941811). Author contributions Junqi Pang conducted data curation, formal analysis, software, visualization and writing – original draft. Yuguang Zhang conducted software and supervision. Keyang Sun analyzed texture properties.Wanyue Jiang and Xuesi Pan conducted data curation. Qingyu Yang and Yuzhe Gao aided in the conceptualization of the work, project administration, funding acquisition and project administration. Competing interests The authors declare no competing interests. References Huang, L., Wang, T., Han, Z., Meng, Y. & Lu, X. Effect of egg yolk freezing on properties of mayonnaise. 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Cite Share Download PDF Status: Published Journal Publication published 12 Jan, 2026 Read the published version in npj Science of Food → Version 1 posted Editorial decision: Revision requested 15 Nov, 2025 Reviews received at journal 15 Nov, 2025 Reviewers agreed at journal 13 Nov, 2025 Reviewers agreed at journal 13 Nov, 2025 Reviews received at journal 12 Nov, 2025 Reviewers agreed at journal 11 Nov, 2025 Reviewers agreed at journal 11 Nov, 2025 Reviewers agreed at journal 11 Nov, 2025 Reviewers agreed at journal 11 Nov, 2025 Reviewers invited by journal 11 Nov, 2025 Editor assigned by journal 15 Oct, 2025 Submission checks completed at journal 08 Oct, 2025 First submitted to journal 22 Sep, 2025 You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. 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12:47:38","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":285250,"visible":true,"origin":"","legend":"\u003cp\u003eThe RBP was heated at 90℃ for different times under pH 2.0 conditions to form PFs stabilized mayonnaise-like and mayonnaise for characterization. a AP% and b WHC. Different small letters in the same column indicate a significant difference (\u003cem\u003ep\u003c/em\u003e \u0026lt; 0.05).\u003c/p\u003e","description":"","filename":"2.png","url":"https://assets-eu.researchsquare.com/files/rs-7678095/v1/184051c187d43c3d452766ec.png"},{"id":96383955,"identity":"b04968ab-c598-4419-89ab-55241f1f017d","added_by":"auto","created_at":"2025-11-20 12:47:39","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":187093,"visible":true,"origin":"","legend":"\u003cp\u003eRheological properties of the RBP was heated at 90℃ for different times under pH 2.0 conditions to form PFs stabilized mayonnaise-like and mayonnaise. a the curve of G' and G'' variation with scanning frequency and b the curve of viscosity variation with scanning rate.\u003c/p\u003e","description":"","filename":"3.png","url":"https://assets-eu.researchsquare.com/files/rs-7678095/v1/65432b67dbe6e68784cddb17.png"},{"id":96383953,"identity":"1c308978-ce59-4e70-ae9d-4d9d6a82df84","added_by":"auto","created_at":"2025-11-20 12:47:38","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":237056,"visible":true,"origin":"","legend":"\u003cp\u003eVisual appearance and color of RBP was heated at 90℃ for different times under pH 2.0 conditions to form PFs stabilized mayonnaise-like and mayonnaise.Values are mean ± SD. Means with different letters a–l within the same color parameter are significantly different (P \u0026lt; 0.05).\u003c/p\u003e","description":"","filename":"4.png","url":"https://assets-eu.researchsquare.com/files/rs-7678095/v1/7e56f37c452b0e42744d867c.png"},{"id":96452904,"identity":"328fc7a0-1325-4575-ae05-96715fb7da9c","added_by":"auto","created_at":"2025-11-21 09:53:23","extension":"png","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":140917,"visible":true,"origin":"","legend":"\u003cp\u003eSensory evaluation radar chart of RBP was heated at 90℃ for different times under pH 2.0 conditions to form PFs stabilized mayonnaise-like and mayonnaise.\u003c/p\u003e","description":"","filename":"5.png","url":"https://assets-eu.researchsquare.com/files/rs-7678095/v1/ebc896893f030abebd63ee00.png"},{"id":96454308,"identity":"8f4f10fd-d218-46d8-a394-70f49a777ad1","added_by":"auto","created_at":"2025-11-21 10:02:36","extension":"png","order_by":6,"title":"Figure 6","display":"","copyAsset":false,"role":"figure","size":139245,"visible":true,"origin":"","legend":"\u003cp\u003eStorage Quality Properties of the Mayonnaise-like emulsions at Room Temperature. a peroxide value, b MDA content and c colonies number. Different capital letters indicate significant differences (\u003cem\u003ep \u003c/em\u003e\u0026lt; 0.05)among the same samples at different times. Different lowercase letters indicate a significant difference (\u003cem\u003ep\u003c/em\u003e\u0026lt; 0.05) among different samples on the same day. The same notation applies below.\u003c/p\u003e","description":"","filename":"6.png","url":"https://assets-eu.researchsquare.com/files/rs-7678095/v1/e82c04fb7df1d6b9a644c26e.png"},{"id":100614490,"identity":"ede30632-7bcf-48de-8b0b-622f217e5f5f","added_by":"auto","created_at":"2026-01-19 17:20:37","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":2206553,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-7678095/v1/8d2cc733-e941-45ca-804d-73f61be71b9b.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"Edible mayonnaise-like emulsion stabilized by rice bran protein fibril aggregation: Effect of fibril aggregate structure","fulltext":[{"header":"1. Introduction","content":"\u003cp\u003eMayonnaise is a widely used condiment in many dishes. However, its health and application drawbacks have recently attracted consumer attention\u003csup\u003e\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e\u003c/sup\u003e. The main concerns include the following: first, its high caloric and fat content, as mayonnaise is primarily composed of oil and egg yolks, which can contribute to obesity and related health issues\u003csup\u003e\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e\u003c/sup\u003e; second, its cholesterol and sodium levels, which, with long-term excessive intake, may affect blood pressure and cardiovascular health and are unsuitable for individuals with high cholesterol\u003csup\u003e\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e\u003c/sup\u003e; and third, the presence of egg components, which makes it inappropriate for individuals with egg allergies\u003csup\u003e\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e\u003c/sup\u003e. Because of these limitations, some consumers prefer healthier alternatives or low-fat versions to better align with their dietary and health needs\u003csup\u003e\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e\u003c/sup\u003e. Consequently, innovating formulations to create healthier and more diverse mayonnaise products has become a significant goal for industry development\u003csup\u003e6\u003c/sup\u003e.\u003c/p\u003e\u003cp\u003ePlant-based mayonnaise products have gained popularity due to their environmental, health, and sustainability benefits. Compared with traditional animal-based mayonnaise, their production generally requires less carbon and water, making them more environmentally friendly\u003csup\u003e\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e7\u003c/span\u003e\u003c/sup\u003e. Moreover, plant proteins are cholesterol-free and rich in unsaturated fatty acids, making them suitable for formulating low-cholesterol mayonnaise. From a sourcing perspective, plant proteins are more widely available and can contribute to agricultural development\u003csup\u003e\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e\u003c/sup\u003e. Commonly used plant proteins in mayonnaise production include pea, soy, and chickpea proteins. However, not all plant proteins are suitable for mayonnaise-like products, as they must exhibit strong emulsifying and textural properties to replicate the texture and mouthfeel of traditional mayonnaise. To address this, the modification of plant proteins to improve their emulsifying functionality is a common industrial strategy\u003csup\u003e9\u003c/sup\u003e.\u003c/p\u003e\u003cp\u003eRice bran protein (RBP), extracted from rice bran (a byproduct of rice milling with a protein content of 10%\u0026ndash;16%), is typically obtained using an alkali-soluble acid-precipitation method from defatted rice bran. Its unique amino acid composition imparts nutritional value and potential for industrial applications\u003csup\u003e\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e10\u003c/span\u003e\u003c/sup\u003e. RBP is a low-allergenic, high-quality plant protein with desirable functional properties such as solubility, emulsification, and foaming\u003csup\u003e\u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e\u003c/sup\u003e. However, its native structure limits its ability to meet diverse market demands, necessitating modifications to enhance its functionality\u003csup\u003e\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e,\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e\u003c/sup\u003e. In recent years, various physical, chemical, and enzymatic modification strategies have been widely employed for this purpose\u003csup\u003e14\u003c/sup\u003e. Among these, acid-thermal treatment of RBP to generate fibril structures has proven particularly effective and desirable for practical applications. The resulting fibrils, characterized by abundant cross-β-sheets and \u0026ldquo;zipper-like\u0026rdquo; configurations, provide excellent stability under extreme conditions. This treatment improves multiple functional properties of RBP, including viscosity, gelation, emulsification, foaming, antioxidant activity, and antimicrobial activity, highlighting its significant potential for future applications in food processing\u003csup\u003e\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e\u003c/sup\u003e.\u003c/p\u003e\u003cp\u003eProtein modification to form fiber aggregates enhances the functional properties of proteins. Compared with native proteins, fiber aggregates show significant improvements in gelation, emulsifying activity, and foaming properties\u003csup\u003e\u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e\u003c/sup\u003e. The superior gelation capacity of protein fiber (PF) aggregates results from their ability to cross-link with polysaccharides in aqueous environments, forming hydrogels with strong water retention and elasticity. These properties enable their use in food applications as thickeners to achieve desired consistency and texture\u003csup\u003e\u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e\u003c/sup\u003e. PF aggregates also exhibit enhanced emulsifying and foaming performance. Owing to their higher aspect ratio, fiber structures demonstrate superior emulsifying effects and are widely used in the preparation of high internal phase emulsions\u003csup\u003e18\u003c/sup\u003e. Studies have further shown that PFs can stabilize Pickering emulsions through irreversible interfacial adsorption and anti-aggregation properties while improving the bioavailability of encapsulated nutrients. Additionally, surface hydrophobicity has been identified as a key factor influencing the emulsifying and foaming behavior of fiber aggregates\u003csup\u003e\u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e19\u003c/span\u003e\u003c/sup\u003e. Importantly, the quality and stability of plant-based mayonnaise products are directly linked to the emulsifying and gelation properties of the proteins employed\u003csup\u003e\u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e20\u003c/span\u003e\u003c/sup\u003e. Thus, developing mayonnaise-like products using plant PF aggregates holds significant potential.\u003c/p\u003e\u003cp\u003eHerein, RBP aggregates with different structures, prepared by heating them for varying durations at pH 2.0, were used to develop mayonnaise-like products. The properties of these products, including rheology, color differences, interfacial protein adsorption (AP%), texture, water-holding capacity (WHC), sensory attributes, and storage stability, were evaluated to assess the performance of mayonnaise-like products formulated with RBP fiber structures. The results were then compared with those of traditional egg yolk\u0026ndash;based mayonnaise to verify the feasibility and practicality of using fiber-structured RBP in the production of mayonnaise-like products.\u003c/p\u003e"},{"header":"2. Methods","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e\n \u003ch2\u003e2.1 Materials\u003c/h2\u003e\n \u003cp\u003eDefatted rice bran was provided by Shenyang Aoda Oil (Liaoning, China).Fresh eggs were purchased from the local market (Shenyang, China). Soybean oil was provided by Yili Goli Company (Shanghai, China). Salt was provided by Yunnan Salt Industry Co., Ltd (Yunnan, China). White sugar and White vinegar were provided by Lianhua Health Industry Group Co., Ltd (Henan, China). Anhydrous ethanol, sodium hydroxide and hydrochloric acid were provided by Tianjin Hengxing Chemical Reagent Manufacturing Co., Ltd. (Tianjin, China). Isooctane, Isopropanol, N-butanol, Trichloroacetic acid, Thiobarbituric acid, Potassium thiocyanate, Ferrous sulfate, Barium chloride, 1,1,3,3-tetraethoxypropane and Normal saline all purchased from Shanghai Aladdin Biochemical Technology Co., Ltd. (Shanghai, China), all the purities are at the analytical purity level. All aqueous solutions were prepared using distilled water.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec4\" class=\"Section2\"\u003e\n \u003ch2\u003e2.2 Experimental Method\u003c/h2\u003e\n \u003cdiv id=\"Sec5\" class=\"Section3\"\u003e\n \u003ch2\u003e2.2.1 Protein extraction\u003c/h2\u003e\n \u003cp\u003eA previously described method\u003csup\u003e\u003cspan class=\"CitationRef\"\u003e21\u003c/span\u003e\u003c/sup\u003e was followed with slight modifications. Defatted rice bran was mixed with 10 volumes of distilled water. The pH was adjusted to 9.5 with 1.0 mol/L NaOH. The mixture was stirred at 45 ℃ for 2 h, and then centrifuged at a speed of 4000 r/min for 15 min using the centrifuge (LD4-2, Beckmann Kurtz Trading Company, USA). The supernatant was collected and adjusted to pH 4.5 with 1.0 mol/L HCl, followed by centrifugation at 4000 r/min for 15 min. The precipitate was adjusted to a neutral pH. The mixture was then freeze-dried using the freeze-drying machine (SCIENTZ-12NIA, Ningbo Xinzi Biotechnology Co., Ltd., China). The protein content of the freeze-dried powder was determined using the Kjeldahl method.\u003c/p\u003e\n \u003c/div\u003e\n \u003cdiv id=\"Sec6\" class=\"Section3\"\u003e\n \u003ch2\u003e2.2.2 Preparation of protein fiber polymers\u003c/h2\u003e\n \u003cp\u003eA 10 mg/mL RBP solution was prepared and stirred at 40\u0026deg;C for 2 h to ensure complete dissolution. The pH was adjusted to 2.0, and the solution was then heated at 90\u0026deg;C for varying durations (0, 2, 4, 6, 8, 10, and 12 h). After heating, the samples were immediately cooled in an ice-water bath to obtain protein solutions at several stages of the PF preparation process.\u003c/p\u003e\n \u003c/div\u003e\n \u003cdiv id=\"Sec7\" class=\"Section3\"\u003e\n \u003ch2\u003e2.2.3 Atomic Force Microscopy (AFM) images\u003c/h2\u003e\n \u003cp\u003eThe RBP samples were diluted to a concentration of 0.02 mg/mL with distilled water (pH 2.0). Subsequently, 5 \u0026micro;L of the diluted sample was deposited onto a mica surface and dried at 25\u0026deg;C for 24 h. AFM images were taken using a Dimension Icon AFM with an SNL-10 cantilever probe in tapping mode (Bruker Corporation, Billerica, MA, USA). AFM images were analyzed with NanoScope Analysis version 1.90 and FiberApp software. The dimensions of the fibrils were measured using the cross-sectional tool in NanoScope Analysis version 1.90. FiberApp software, which is designed to track and analyze the contour lengths of flexible and rigid fibrils, was used to make the calculations.\u003c/p\u003e\n \u003c/div\u003e\n \u003cdiv id=\"Sec8\" class=\"Section3\"\u003e\n \u003ch2\u003e2.2.4 Preparation of PFs stabilized Mayonnaise-like emulsions\u003c/h2\u003e\n \u003cp\u003e\u003cstrong\u003eThe traditional mayonnaise preparation was prepared as follows\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003e5.0 mL of egg yolk was weighed into a clean 50-mL centrifuge tube. 0.2 g salt, 0.4 g white sugar, and 0.5 g white vinegar were added. The mixture was stirred until the salt and sugar were completely dissolved. Subsequently, 15.0 mL soybean oil was added to the centrifuge tube, and the mixture was homogenized at 16000 r/min for 2 min.\u003c/p\u003e\n \u003cp\u003ePF-stabilized mayonnaise was prepared as follows: 5.0 mL PF was prepared by heating at pH 2.0 (10 mg/mL) and was weighed into a clean 50-mL centrifuge tube. 0.2 g salt, 0.4 g white sugar, and 0.5 g white vinegar were added, and the mixture was thoroughly stirred until the salt and sugar were dissolved. Subsequently, 15.0 g soybean oil was added to the centrifuge tube. The mixture was homogenized at 16000 r/min for 2 min.\u003c/p\u003e\n \u003c/div\u003e\n \u003cdiv id=\"Sec9\" class=\"Section3\"\u003e\n \u003ch2\u003e2.2.5 Protein adsorption (AP%)\u003c/h2\u003e\n \u003cp\u003eThe AP% was measured using a previously described method\u003csup\u003e22\u003c/sup\u003e. A 20 mL aliquot of the freshly prepared mayonnaise-like product was centrifuged at 4000 r/min for 10 min. The lower transparent layer of the separated liquid was gently aspirated. Its protein concentration was determined using the Coomassie Brilliant Blue method. The AP% of the mayonnaise-like product was then calculated as follows:\u003c/p\u003e\n \u003cdiv id=\"Equa\" class=\"Equation\"\u003e\n \u003cdiv class=\"mathdisplay\" id=\"FileID_Equa\" name=\"EquationSource\"\u003e$$\\:AP\\%=\\frac{{C}_{S}-{C}_{f}}{{C}_{s}}\\times\\:100$$\u003c/div\u003e\n \u003c/div\u003e\n \u003cp\u003eIn the formula, \u003cem\u003eC\u003c/em\u003e\u003csub\u003e\u003cem\u003es\u003c/em\u003e\u003c/sub\u003e represents the total protein content in the mayonnaise-like product (mg/mL), and \u003cem\u003eC\u003c/em\u003e\u003csub\u003e\u003cem\u003ef\u003c/em\u003e\u003c/sub\u003e indicates the protein content in the aqueous phase after centrifugation (mg/mL).\u003c/p\u003e\n \u003c/div\u003e\n \u003cdiv id=\"Sec10\" class=\"Section3\"\u003e\n \u003ch2\u003e2.2.6 Water Holding Capacity (WHC)\u003c/h2\u003e\n \u003cp\u003eThe prepared mayonnaise-like emulsions was transferred into a 50mL centrifuge tube and weighed to determine the initial mass (m\u003csub\u003e1\u003c/sub\u003e). The sample was then centrifuged at 6000 r/min for 5 min. The water separated in the centrifuge tube was removed, and the tube was weighed again to find the final mass (m\u003csub\u003e2\u003c/sub\u003e). The WHC of the mayonnaise-like product was calculated using the following formula:\u003c/p\u003e\n \u003cp\u003e\u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(\\:\\text{W}\\text{H}\\text{C}（\\text{%}）=\\frac{{\\text{m}}_{2}-{\\text{m}}_{0}}{{\\text{m}}_{1}-{\\text{m}}_{0}}\\times\\:\\)\u003c/span\u003e\u003c/span\u003e100 \u003csup\u003e23\u003c/sup\u003e\u003c/p\u003e\n \u003cp\u003eIn the formula, m\u003csub\u003e0\u003c/sub\u003e is the mass of the centrifuge tube, m\u003csub\u003e1\u003c/sub\u003e is the total mass of the centrifuge tube and egg yolk sauce before centrifugation, and m\u003csub\u003e2\u003c/sub\u003e represents the total mass of the centrifuge tube and egg yolk sauce after centrifugation.\u003c/p\u003e\n \u003c/div\u003e\n \u003cdiv id=\"Sec11\" class=\"Section3\"\u003e\n \u003ch2\u003e2.2.7 Rheological properties\u003c/h2\u003e\n \u003cp\u003eThe rheological properties of the mayonnaise-like emulsions were immediately examined after preparation with a hybrid rheometer (DHR-1, 10 nN\u0026middot;m-50 mN\u0026middot;m; TA Instruments, New Castle, DE, USA) to measure both dynamic and steady-state shear rheological properties. Measurements were taken using parallel plate geometry with a diameter of 40 mm and a gap of 1000 \u0026micro;m. First, the mayonnaise-like product viscosity was assessed by flow sweep mode at shear rates ranging from 0.01 to 100 s\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e at a constant temperature of 25\u0026deg;C, and the viscosity ratio was calculated over a range of 0.1\u0026ndash;100 s.\u003c/p\u003e\n \u003cp\u003eFor the dynamic oscillation scanning tests, measurements were also taken with a parallel plate with a diameter of 40 mm and a gap of 1000 \u0026micro;m. The tested emulsion was placed on the platform, and the plate was pressed down to the trim distance. Excess sample around the plate was removed. The sample was sealed with silicone oil. The storage modulus (G\u0026apos;) and loss modulus (G\u0026apos;\u0026apos;) of the mayonnaise-like product were determined at a strain value of 0.2% and a temperature of 25\u0026deg;C using the angular frequency sweep mode.\u003c/p\u003e\n \u003c/div\u003e\n \u003cdiv id=\"Sec12\" class=\"Section3\"\u003e\n \u003ch2\u003e2.2.8 Texture properties\u003c/h2\u003e\n \u003cp\u003eThe texture properties of the mayonnaise-like emulsions were evaluated using an established method\u003csup\u003e\u003cspan class=\"CitationRef\"\u003e24\u003c/span\u003e\u003c/sup\u003e, with slight modifications. The samples were prepared in 50mL centrifuge tubes and subjected to texture analysis at 25\u0026deg;C with a TA-XT Plus texture analyzer equipped with a P/36R cylindrical probe. Hardness, adhesiveness, cohesiveness, springiness, and chewiness were assessed. The measurement parameters were set as follows: the pre-test speed was 1.0 mm/s, the test speed was 0.6 mm/s, the post-test speed was 1.0 mm/s, the deformation degree was 30%, and the trigger force was 0.3 g. Each sample was tested in triplicate, and the average values are reported.\u003c/p\u003e\n \u003c/div\u003e\n \u003cdiv id=\"Sec13\" class=\"Section3\"\u003e\n \u003ch2\u003e2.2.9 Color difference\u003c/h2\u003e\n \u003cp\u003e10 mL of the freshly prepared mayonnaise and mayonnaise-like emulsions were placed into display bottles. The colorimeter was calibrated using a white calibration plate. The L* (lightness), a* (redness), and b* (yellowness) values were measured for each sample. Each sample was measured in triplicate. The average values were recorded.\u003c/p\u003e\n \u003c/div\u003e\n \u003cdiv id=\"Sec14\" class=\"Section3\"\u003e\n \u003ch2\u003e2.2.10 Sensory evaluation\u003c/h2\u003e\n \u003cp\u003eSensory evaluation of the mayonnaise products was performed by a semi-trained panel of 10 food science students. The panel assessed the color, consistency, aroma, flavor, and overall acceptability of the samples. The average score assigned by the panelists for each attribute served as the evaluation score for that attribute. The scoring criteria were as follows: each attribute could be given a maximum of 20 points. Scores ranging from 17 to 20 indicated high satisfaction with the product, whereas scores between 12 and 16 indicated moderate satisfaction, and scores below 12 indicated dissatisfaction with the product quality.\u003c/p\u003e\n \u003c/div\u003e\n \u003cdiv id=\"Sec15\" class=\"Section3\"\u003e\n \u003ch2\u003e2.2.11 Storage Quality Properties of the Mayonnaise-like emulsions at Room Temperature\u003c/h2\u003e\n \u003cp\u003eThe storage stability of mayonnaise and the mayonnaise-like product was evaluated using a room-temperature test. Freshly prepared mayonnaise and the mayonnaise-like product were stored at room temperature for 30 d. Measurements were taken every 3 d, with three samples drawn each time and testing performed in triplicate. Indicators tested included peroxide value, malondialdehyde (MDA) content, and total bacterial count.\u003c/p\u003e\n \u003c/div\u003e\n \u003cdiv id=\"Sec16\" class=\"Section3\"\u003e\n \u003ch2\u003e2.2.11.1 Peroxide Value\u003c/h2\u003e\n \u003cp\u003eThe peroxide value determination in mayonnaise was based on the standard GB 5009.227\u0026ndash;2023. The specific procedure was as follows: 50 \u0026micro;L freshly prepared mayonnaise and mayonnaise-like product prepared with PF were accurately dispensed into a clean 5mL centrifuge tube. 3 mL heated iso-octane/isopropanol (3:1, V/V) was added, and the samples were thoroughly mixed using a vortex mixer. The mixture was centrifuged at 8000 r/min for 5 min. 0.2 mL of the supernatant was mixed with 2.8 mL methanol/n-butanol (2:1, V/V). 0.15 mL freshly prepared FeCl\u003csub\u003e2\u003c/sub\u003e solution (0.25 mol/L) and 0.15 mL potassium thiocyanate solution (0.3 g/mL) were added. The mixture was allowed to react in the dark for 20 min. Measure the absorbance using an ultraviolet spectrophotometer (UV-1200S, Aowei Instrument, China) and the absorbance was measured at 510 nm. The measured absorbance values were used to calculate the peroxide value of the mayonnaise by referencing the standard curve established with Fe\u003csup\u003e3+\u003c/sup\u003e solutions. The standard curve was established using Fe\u003csup\u003e3+\u003c/sup\u003e solutions with concentrations of 0.02, 0.04, 0.08, 0.16, and 0.20 mmol/L using a methanol:n-butanol (2:1, V/V) mixture as the blank. The equation of the standard curve is y\u0026thinsp;=\u0026thinsp;4.50312x \u0026minus;\u0026thinsp;0.0081 (R\u0026sup2; = 0.9905). Each sample was measured in triplicate, and the average value was calculated.\u003c/p\u003e\n \u003c/div\u003e\n \u003cdiv id=\"Sec17\" class=\"Section3\"\u003e\n \u003ch2\u003e2.2.11.2 MDA content\u003c/h2\u003e\n \u003cp\u003eMDA content in mayonnaise was measured in accordance with GB 5009.181\u0026ndash;2016. The standard curve was generated using 1,1,3,3-tetraethoxypropane, according to the equation y\u0026thinsp;=\u0026thinsp;1.04561x\u0026thinsp;+\u0026thinsp;0.0035 (R\u003csup\u003e2\u003c/sup\u003e\u0026thinsp;=\u0026thinsp;0.9991).\u003c/p\u003e\n \u003c/div\u003e\n \u003cdiv id=\"Sec18\" class=\"Section3\"\u003e\n \u003ch2\u003e2.2.11.3 Total viable count\u003c/h2\u003e\n \u003cp\u003eThe determination of the total viable count (TVC) in mayonnaise during storage was performed in accordance with GB 4789.2\u0026ndash;2022, the National Food Safety Standard for Microbiological Examination of Food, specifically the method for total viable count determination.\u003c/p\u003e\n \u003c/div\u003e\n \u003cdiv id=\"Sec19\" class=\"Section3\"\u003e\n \u003ch2\u003e2.2.12 Statistical analysis\u003c/h2\u003e\n \u003cp\u003eAll measurement results were tested three times, from which the average value and standard deviation value were calculated. The statistical differences, mean values and standard deviations of the data were analyzed using the SPSS 24.0 software package. Furthermore, multiple comparisons were conducted using the Tukey test. Different letters indicate p\u0026thinsp;\u0026lt;\u0026thinsp;0.05. Origin 2022 is used to complete the drawings.\u003c/p\u003e\n \u003c/div\u003e\n\u003c/div\u003e"},{"header":"3. Results and Discussion","content":"\u003cdiv id=\"Sec21\" class=\"Section2\"\u003e\n \u003ch2\u003e3.1 AFM images of PFs\u003c/h2\u003e\n \u003cp\u003eAFM is a high-resolution microscopic technique enabling direct observation of the microscopic morphology of RBP fiber aggregates\u003csup\u003e\u003cspan class=\"CitationRef\"\u003e25\u003c/span\u003e\u003c/sup\u003e. AFM images were used to study the microscopic morphological changes of RBP heated for 0, 6, and 12 h at pH 2.0.\u003c/p\u003e\n \u003cp\u003eThe results are shown in Fig. \u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003e. At pH 2.0, PFs formed after heating for 0 h presented large spherical particles with a chain width of approximately 300 nm and a chain height of approximately 23 nm. The overly large particle structure was not suitable as an emulsifier at the oil-water interface. After 12 h of heating at pH 2.0, PFs formed, and multiple PFs with slender fiber structures could be seen in the field of view. Under other heating times, these structures were not clearly visible. Compared to the PFs formed after heating for 6 h, the chain width and chain height of these PFs increased by 25.0% and 36.5%, respectively. The larger chain width and chain height given the mayonnaise-like emulsions stabilized from PFs better stability\u003csup\u003e\u003cspan class=\"CitationRef\"\u003e26\u003c/span\u003e\u003c/sup\u003e. This result indicated that chose the appropriate heating time was crucial for the formation of RBP fibers. The heating time changed the secondary structure and surface charge state of the protein, thereby affected the intermolecular interactions and promoting the formation of fibers\u003csup\u003e27\u003c/sup\u003e.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec22\" class=\"Section2\"\u003e\n \u003ch2\u003e3.2 AP% analysis\u003c/h2\u003e\n \u003cp\u003eIn emulsion systems, the level of AP% has a significant impact on the stability and functional properties of the emulsion\u003csup\u003e\u003cspan class=\"CitationRef\"\u003e28\u003c/span\u003e\u003c/sup\u003e. The AP% of the mayonnaise-like emulsion made with RBP and the mayonnaise prepared with egg yolk are shown in Fig. \u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003e-a.\u003c/p\u003e\n \u003cp\u003eThe homemade mayonnaise made with egg yolk exhibited the poorest performance in terms of AP%. This is attributed to its larger droplet size and less dense distribution of the emulsion droplets in mayonnaise prepared by egg yolk homogenization. Conversely, the mayonnaise-like emulsion prepared with RBP aggregates heated for different times at pH 2.0 showed no significant difference in AP%. Only the mayonnaise-like emulsion prepared with the native protein without heating exhibited significantly lower AP% because the hydrophobic groups in the native protein are protected within the protein matrix and are not exposed\u003csup\u003e\u003cspan class=\"CitationRef\"\u003e29\u003c/span\u003e\u003c/sup\u003e. After heating for 12 h, RBP forms elongated fibrous structures. This process modifies the native RBP, altering its surface structure and chemical properties, thereby increasing its adsorption at the oil\u0026ndash;water interface in the mayonnaise-like emulsion\u003csup\u003e30\u003c/sup\u003e. Changes in the surface charge distribution of the protein accompany the formation of fibrous structures. If the surface carries more charge that can interact with the oil\u0026ndash;water interface, its adsorption capacity is enhanced\u003csup\u003e\u003cspan class=\"CitationRef\"\u003e31\u003c/span\u003e\u003c/sup\u003e. Additionally, studies have shown that protein fibrous structure formation reduces the molecular weight of the protein. Smaller molecular weight fragments may more easily diffuse to and adsorb at the oil\u0026ndash;water interface\u003csup\u003e\u003cspan class=\"CitationRef\"\u003e32\u003c/span\u003e\u003c/sup\u003e.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec23\" class=\"Section2\"\u003e\n \u003ch2\u003e3.3 WHC analysis\u003c/h2\u003e\n \u003cp\u003eWHC is one of the fundamental characteristics for evaluating the structural stability of emulsions. It reflects the ability of the emulsion system to retain water\u003csup\u003e\u003cspan class=\"CitationRef\"\u003e33\u003c/span\u003e\u003c/sup\u003e. The WHC of various mayonnaise samples is shown in Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003e-b. The mayonnaise prepared with egg yolk showed the poorest WHC. This is because the aqueous phase is relatively abundant during mayonnaise preparation, and the proteins and lecithin in the egg yolk alone cannot stabilize the mayonnaise emulsion. Among the mayonnaise-like emulsions prepared with RBP, the one with the highest WHC is the emulsion stabilized by PF obtained after heating for 12 h at pH 2.0, because the fibrillar proteins possess a highly ordered \u0026beta;-sheet structure. This tightly packed fibrous morphology provides a large surface area and a multipoint hydrogen-bonding network, facilitating water capture and retention. Additionally, the fibers form larger aggregates, which can adsorb more water, enhancing WHC\u003csup\u003e\u003cspan class=\"CitationRef\"\u003e34\u003c/span\u003e\u003c/sup\u003e. In contrast, the native structure of RBP restricts its interaction with water molecules, resulting in a poorer WHC.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec24\" class=\"Section2\"\u003e\n \u003ch2\u003e3.4 Rheological properties analysis\u003c/h2\u003e\n \u003cp\u003eThe droplet aggregation state in mayonnaise-like emulsions determines their rheological properties. As shown in Fig. \u003cspan class=\"InternalRef\"\u003e4\u003c/span\u003e, not all mayonnaise-like emulsions stabilized with PFs are supported viscoelastic emulsions that can maintain sharp edges. Only the PFs stabilized mayonnaise-like emulsions formed after heating at pH 2.0 and 90 ℃ for 8 h showed a viscoelastic emulsion with a supporting structure. Moreover, the longer the heating time, the better the edge retention. As shown in Fig. \u003cspan class=\"InternalRef\"\u003e3\u003c/span\u003e-a, all the mayonnaise-like emulsions samples exhibited G\u0026apos; \u0026gt;G\u0026quot;, indicating that the emulsion was mainly elastic\u003csup\u003e\u003cspan class=\"CitationRef\"\u003e35\u003c/span\u003e\u003c/sup\u003e. In the RBP heated to form PFs stabilized mayonnaise-like emulsions at pH 2.0, the sample group heated for 12 h exhibited the best rheological properties, with G\u0026apos; and G\u0026quot; higher than those of other samples. Combined with AFM image analysis, this is because the PFs can form a network structure, better adsorbing on the oil-water interface, making the mayonnaise-like emulsions exhibit more stable properties\u003csup\u003e\u003cspan class=\"CitationRef\"\u003e25\u003c/span\u003e\u003c/sup\u003e. Therefore, acid heating modification of RBP to obtain a performance suitable for industrial production of egg yolk sauce analogues has significant commercial value.\u003c/p\u003e\n \u003cp\u003eAs the scanning frequency increased, the G\u0026apos; and G\u0026quot; values of all sample groups except the PFs-0 h stabilized mayonnaise-like emulsions remain relatively stable. This once again indicates that the large particles of protein, as emulsifiers, have poor stability. When affected by external forces, they are prone to undergo undesirable changes in properties, which is not conducive to industrial production. The frequency-independent rheological behavior of PFs prepared under pH 2.0 conditions can be attributed to their high protein adsorption on the emulsion interfacial membrane, which enhanced the interaction between droplets.\u003c/p\u003e\n \u003cp\u003eThe curves showed the viscosity changes of mayonnaise and PFs stabilized mayonnaise-like emulsions with respect to shear rate are shown in Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e3\u003c/span\u003e-b. All samples exhibited shear thinning behavior, which was explained by the coagulation of emulsion droplets\u003csup\u003e36\u003c/sup\u003e. However, among these samples, the mayonnaise-like emulsions of PFs-12 h that was stable showed the highest viscosity at the beginning of shear. This was due to the fibrous structure of PFs-12 h, which allowed more protein adsorption at the emulsion interface. As the heating time increased, the RBP structure unfolded and exposed hydrophobic groups, thereby promoting the formation of smaller droplets during the emulsion preparation process and causing a more compact droplet arrangement\u003csup\u003e\u003cspan class=\"CitationRef\"\u003e37\u003c/span\u003e\u003c/sup\u003e.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec25\" class=\"Section2\"\u003e\n \u003ch2\u003e3.5 Texture properties analysis\u003c/h2\u003e\n \u003cp\u003eThe textural properties of mayonnaise are an essential factor influencing its sensory characteristics and consumer acceptance. Texture profile analysis, conducted using a texture analyzer, involves two compressions of the samples to measure parameters such as hardness, adhesiveness, springiness, and cohesiveness. The textural properties of the mayonnaise-like emulsions prepared from RBP aggregates and the homemade mayonnaise are shown in Table 1.\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003eTable.1 Texture properties of different PFs mayonnaise-like and mayonnaise\u003c/strong\u003e\u003c/p\u003e\n \u003ctable id=\"Taba\" border=\"1\"\u003e\n \u003cthead\u003e\n \u003ctr\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eCondition\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eHeating time/h\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eHardness/N\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eViscosity/mj\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eCohesion\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eElasticity/mm\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003c/thead\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" rowspan=\"7\"\u003e\n \u003cp\u003epH\u0026thinsp;=\u0026thinsp;2.0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e3.2\u0026thinsp;\u0026plusmn;\u0026thinsp;0.2\u003csup\u003ei\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.61\u0026thinsp;\u0026plusmn;\u0026thinsp;0.05\u003csup\u003ej\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.65\u0026thinsp;\u0026plusmn;\u0026thinsp;0.04\u003csup\u003ehi\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.72\u0026thinsp;\u0026plusmn;\u0026thinsp;0.03\u003csup\u003eij\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e3.45\u0026thinsp;\u0026plusmn;\u0026thinsp;0.09\u003csup\u003ehi\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.83\u0026thinsp;\u0026plusmn;\u0026thinsp;0.05\u003csup\u003egh\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.75\u0026thinsp;\u0026plusmn;\u0026thinsp;0.03\u003csup\u003efg\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.85\u0026thinsp;\u0026plusmn;\u0026thinsp;0.04\u003csup\u003ehi\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e3.79\u0026thinsp;\u0026plusmn;\u0026thinsp;0.03\u003csup\u003egh\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.93\u0026thinsp;\u0026plusmn;\u0026thinsp;0.03\u003csup\u003eg\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.81\u0026thinsp;\u0026plusmn;\u0026thinsp;0.02\u003csup\u003ef\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1\u0026thinsp;\u0026plusmn;\u0026thinsp;0.04\u003csup\u003eg\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e4.38\u0026thinsp;\u0026plusmn;\u0026thinsp;0.19\u003csup\u003eef\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.08\u0026thinsp;\u0026plusmn;\u0026thinsp;0.03\u003csup\u003ef\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.9\u0026thinsp;\u0026plusmn;\u0026thinsp;0.04\u003csup\u003ee\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.29\u0026thinsp;\u0026plusmn;\u0026thinsp;0.12\u003csup\u003ef\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e5.28\u0026thinsp;\u0026plusmn;\u0026thinsp;0.18\u003csup\u003ed\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.39\u0026thinsp;\u0026plusmn;\u0026thinsp;0.05\u003csup\u003ee\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.97\u0026thinsp;\u0026plusmn;\u0026thinsp;0.03\u003csup\u003ede\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.64\u0026thinsp;\u0026plusmn;\u0026thinsp;0.06\u003csup\u003ee\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e10\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e5.47\u0026thinsp;\u0026plusmn;\u0026thinsp;0.27\u003csup\u003ed\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.63\u0026thinsp;\u0026plusmn;\u0026thinsp;0.04\u003csup\u003ed\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.05\u0026thinsp;\u0026plusmn;\u0026thinsp;0.03\u003csup\u003ed\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.82\u0026thinsp;\u0026plusmn;\u0026thinsp;0.05\u003csup\u003ed\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e12\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e6.28\u0026thinsp;\u0026plusmn;\u0026thinsp;0.22\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.9\u0026thinsp;\u0026plusmn;\u0026thinsp;0.03\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.3\u0026thinsp;\u0026plusmn;\u0026thinsp;0.07\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e2.04\u0026thinsp;\u0026plusmn;\u0026thinsp;0.06\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" colspan=\"2\"\u003e\n \u003cp\u003eegg yolk\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e7.65\u0026thinsp;\u0026plusmn;\u0026thinsp;0.22\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e3.5\u0026thinsp;\u0026plusmn;\u0026thinsp;0.27\u003csup\u003ed\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.81\u0026thinsp;\u0026plusmn;\u0026thinsp;0.04\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e8.32\u0026thinsp;\u0026plusmn;\u0026thinsp;0.18\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003ctfoot\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"6\"\u003eNote: Different small letters in the same column indicate a significant difference (\u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.05).\u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tfoot\u003e\n \u003c/table\u003e\n\u003c/div\u003e\n\u003cp\u003eMayonnaise prepared from egg yolk demonstrated superior textural properties, attributed to the presence of emulsifying components in egg yolk. For the RBP-based mayonnaise-like emulsions, different heating durations at pH 2.0 significantly affected their textural properties. Specifically, the overall textural performance of samples prepared at pH 2.0 was higher than that of mayonnaise prepared with egg yolk. This is because the pH environment affects the protein structure in the solution, causing denaturation, thereby influencing the emulsifying properties of the proteins. Under pH 2.0, the longer the RBP solution was heated, the better the hardness and viscosity of the resulting mayonnaise-like emulsion, and the higher its cohesiveness and springiness\u003csup\u003e\u003cspan class=\"CitationRef\"\u003e23\u003c/span\u003e\u003c/sup\u003e. This trend is consistent with the changes in elasticity and viscosity observed in the rheological measurements.\u003c/p\u003e\n\u003cdiv id=\"Sec26\" class=\"Section2\"\u003e\n \u003ch2\u003e3.6 Color differences analysis\u003c/h2\u003e\n \u003cp\u003eThe color differences of mayonnaise-like emulsions stabilized from RBP heated under pH 2.0 conditions and other mayonnaise samples are shown in Fig. \u003cspan class=\"InternalRef\"\u003e4\u003c/span\u003e.\u003c/p\u003e\n \u003cp\u003eThe L* value, which describes the brightness of a color, indicates how light or dark an object is. The a* value represents how red or green it is, with positive values indicating higher redness and negative values indicating higher greenness. The b* value represents whether the color is more yellow or blue, with positive values indicating higher yellowness and negative values indicating higher blueness. For mayonnaise-like emulsions prepared with RBP, the L* values were all \u0026gt;\u0026thinsp;80, indicating that the emulsions were very bright and close to white. The a* values of all samples were very low; manufacturers typically use this value to help distinguish differences in hue between batches or brands. All mayonnaise samples had relatively high b* values, especially those prepared from egg yolk, due to the inherent color of the egg yolk itself. For RBP-based mayonnaise-like emulsions, there was significant variation in the b* values of emulsions prepared under pH 2.0 conditions. This is because the pH of the protein solution has a substantial impact on the RBP color, which in turn affects the color of the mayonnaise-like emulsion. Specifically, the color of the protein solution becomes darker or lighter at different pH levels, possibly due to protein denaturation or changes in specific amino acid residues. RBP is composed of various amino acids, whose side chains can ionize at different pH levels. For example, glutamic acid and aspartic acid protonate at low pH and deprotonate at high pH. In terms of color acceptability, the mayonnaise-like emulsion prepared from RBP after heating for 12 h at pH 2.0 was more generally accepted. This is particularly important for developing healthier mayonnaise products.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec27\" class=\"Section2\"\u003e\n \u003ch2\u003e3.7 Sensory evaluation analysis\u003c/h2\u003e\n \u003cp\u003eThe sensory evaluation of mayonnaise-like emulsions prepared from RBP aggregates and homemade mayonnaise is shown in Fig. \u003cspan class=\"InternalRef\"\u003e5\u003c/span\u003e.\u003c/p\u003e\n \u003cp\u003eThe sensory scores of the RBP-based mayonnaise-like emulsions were typically higher than those of homemade mayonnaise prepared with egg yolk. For the egg yolk mayonnaise, the lack of additives and the yolk\u0026rsquo;s inherent color and odor contributed to its lower sensory scores. Conversely, for RBP-based mayonnaise-like emulsions, those prepared after heating RBP for 12 h at pH 2.0 received the highest sensory scores for viscosity, because the emulsification process requires homogenization, which involves dispersing oil into fine droplets uniformly distributed in the aqueous phase. Heating RBP for 12 h at pH 2.0 results in the formation of elongated fibrous structures, contributing to this homogenization process.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec28\" class=\"Section2\"\u003e\n \u003ch2\u003e3.8 Determination of Quality and Performance of Mayonnaise Stored at Room Temperature\u003c/h2\u003e\n \u003cdiv id=\"Sec29\" class=\"Section3\"\u003e\n \u003ch2\u003e3.8.1 Peroxide values analysis\u003c/h2\u003e\n \u003cp\u003eThe oxidation of soybean oil in mayonnaise and mayonnaise-like emulsions is a complex process that produces both primary and secondary oxidation products. During the initial oxidation stage, the unsaturated fatty acids in soybean oil\u0026mdash;such as linoleic acid and linolenic acid\u0026mdash;react with oxygen to form peroxides. We measured the oxidation level of mayonnaise and mayonnaise-like emulsions by assessing the primary oxidation product, hydrogen peroxide. Figure \u003cspan class=\"InternalRef\"\u003e6\u003c/span\u003e-a illustrates changes in the hydrogen peroxide content in mayonnaise-like emulsions prepared with RBP over 30 days of storage. As shown in the figure, both freshly prepared egg yolk mayonnaise and mayonnaise-like emulsion samples exhibited high peroxide values (1.54 mmol/kg). This may be due to the high-speed homogenization (16,000 r/min) used in the emulsion preparation process, which increases the emulsion system\u0026rsquo;s temperature and enhances the contact between oxygen and soybean oil, leading to the formation of oxidative products.\u003c/p\u003e\n \u003cp\u003eFigure \u003cspan class=\"InternalRef\"\u003e6\u003c/span\u003e-a demonstrates that among the mayonnaise-like emulsions, the samples prepared with RBP after 12 h of heating had the lowest peroxide values. This is attributed to the high structural stability, strong antioxidant properties, dense network protection, and effective interfacial physicochemical characteristics of the fibrillar protein structure\u003csup\u003e38\u003c/sup\u003e. Therefore, using fibrillar RBP significantly enhances the mayonnaise-like emulsion\u0026rsquo;s stability during storage, extending the product\u0026rsquo;s shelf life, and inhibiting the oxidation of oils.\u003c/p\u003e\n \u003cp\u003eAs shown in Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e6\u003c/span\u003e-a, the peroxide values of mayonnaise-like emulsions made with RBP were significantly lower than those of mayonnaise prepared with egg yolk. The oxidation rate of soybean oil in mayonnaise-like emulsions is affected by several factors: fibrillar protein structure formation in RBP under acidic conditions; the dense droplet structure of mayonnaise-like emulsions prepared at pH 2.0, which reduces the contact between droplets and oxygen, lowering the oxidation rate; and the enhanced activity of certain antioxidants under acidic conditions, which contributes to the antioxidant properties of mayonnaise-like emulsions.\u003c/p\u003e\n \u003c/div\u003e\n \u003cdiv id=\"Sec30\" class=\"Section3\"\u003e\n \u003ch2\u003e3.8.2 MDA content analysis\u003c/h2\u003e\n \u003cp\u003eSoybean oil in mayonnaise oxidizes when exposed to oxygen, producing peroxides. The major oxidation products are mainly hydroperoxides and other primary peroxides, which break down into free radicals under the influence of heat, light, or metal ions, leading to further oxidation. These free radicals react with nearby molecules to form secondary oxidation products like aldehydes (e.g., MDA), carboxylic acids, and ketones, which are typically the sources of negatively perceived flavors. Figure\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e6\u003c/span\u003e-b shows the relationship between MDA production and storage time over 30 d. As shown in the figure, the rate of increase in MDA production was rapid during the first 15 d of storage but slowed over time. This is because MDA is a secondary product formed from primary oxidation products. As the oxidation process continues, the consumption of oxygen slows the primary oxidation process, reducing the primary metabolite hydroperoxide consumption, consequently decreasing secondary oxidation rate.\u003c/p\u003e\n \u003cp\u003eAs shown in Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e6\u003c/span\u003e-b, both RBP-based mayonnaise-like emulsions and homemade mayonnaise exhibited a similar trend in MDA production during storage. However, there were significant differences in the quantities produced. The mayonnaise-like emulsion stabilized by RBP heated for 12 h at pH 2.0 had a MDA content of 1.22 mg/kg, which was significantly lower than that of the other RBP-based emulsions. This trend is consistent with the previously observed changes in peroxide values. This can be attributed to the greater stability of its fibrous RBP structure, resulting in fewer hydroperoxides being generated during storage compared to other samples, reducing the rate of secondary oxidation and leading to lower MDA production.\u003c/p\u003e\n \u003c/div\u003e\n \u003cdiv id=\"Sec31\" class=\"Section3\"\u003e\n \u003ch2\u003e3.8.3 Changes in Total Bacterial Count analysis\u003c/h2\u003e\n \u003cp\u003eTVC determination is a key method for indicating the microbial contamination level in food. A high TVC can indicate the presence of pathogenic bacteria, which poses a threat to consumer health. Changes in the TVC of mayonnaise-like emulsions prepared with heated RBP at pH 2.0 over 30 days of storage are shown in Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e6\u003c/span\u003e-c. Since vinegar was added during mayonnaise preparation, the acetic acid in vinegar lowered the pH of the emulsion system. A low-pH environment is conducive to bacterial growth inhibition. Based on this, the TVC of RBP-based mayonnaise-like emulsions prepared at pH 2.0 should increase less during storage compared to homemade mayonnaise. As shown in Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e6\u003c/span\u003e-c, the measured results are consistent with this prediction. Even after 30 days, the TVC of the mayonnaise-like emulsion remained\u0026thinsp;\u0026lt;\u0026thinsp;900 CFU/ml. Therefore, RBP-based mayonnaise-like emulsions prepared under pH 2.0 conditions do not require preservatives in industrial production. Moreover, the TVC of mayonnaise-like emulsions prepared from fibrillar RBP after 12 h of heating at pH 2.0 is even lower, indicating that this type of emulsion has great potential for developing mayonnaise products.\u003c/p\u003e\n \u003c/div\u003e\n\u003c/div\u003e"},{"header":"Conclusions","content":"\u003cp\u003eThe mayonnaise-like emulsion stabilized by PFs exhibited superior rheological properties with both G\u0026prime; and G\u0026prime;\u0026prime; values higher than those observed under other treatment conditions. This significantly enhanced its shear-thinning behavior and viscoelasticity, as reflected in the high hardness, viscosity, and cohesiveness measured in the textural analysis. The RBP-based emulsions also showed improved color, and in sensory evaluation, the pH 2.0\u0026ndash;treated samples achieved higher scores for consistency and acceptability, attributable to the formation of fibrous structures. These findings are of practical significance for developing healthy, additive-free mayonnaise-like products. Furthermore, the PF-stabilized emulsion prepared under these conditions demonstrated lower MDA production and peroxide values during storage, indicating strong resistance to lipid oxidation. Thus, PF formation contributes to both the stability and desirable characteristics of mayonnaise-like products. Overall, this study provides a valuable reference for the development of plant protein\u0026ndash;based mayonnaise alternatives.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eData availability\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAll data generated or analyzed during this study are included in this published article.\u003c/p\u003e\n\u003cp\u003eReceived: XXX; Accepted: XXX;\u003c/p\u003e\n\u003cp\u003ePublished online:XXX\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAcknowledgments\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis research was supported by Liaoning Province Applied Basic Research Program, (1746603820076); 2025 Liaoning Provincial Department of Education Platform Construction Project (LJ232510166005); Special Plan for Science and Technology Special Envoy Action of Liaoning Provincial Department of Science and Technology (1741935941811).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAuthor contributions\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eJunqi Pang conducted\u0026nbsp;data curation, formal analysis, software, visualization and writing \u0026ndash; original draft.\u0026nbsp;Yuguang Zhang conducted\u0026nbsp;software and supervision. Keyang Sun analyzed texture properties.Wanyue Jiang and Xuesi Pan conducted data curation.\u003c/p\u003e\n\u003cp\u003eQingyu Yang and Yuzhe Gao aided in the conceptualization of the work, project administration, funding acquisition and project administration.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCompeting interests\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors declare no competing interests.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n\u003cli\u003eHuang, L., Wang, T., Han, Z., Meng, Y. \u0026amp; Lu, X. Effect of egg yolk freezing on properties of mayonnaise. \u003cem\u003eFood Hydrocolloids\u003c/em\u003e \u003cstrong\u003e56\u003c/strong\u003e, 311-317 (2016). \u003c/li\u003e\n\u003cli\u003eVieira, M. R., Sim\u0026otilde;es, S., Carrera-S\u0026aacute;nchez, C. \u0026amp; Raymundo, A. 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Droplet size distribution, rheological behavior and stability of corn oil emulsions stabilized by a novel hydrocolloid (Brea gum) compared with gum arabic. \u003cem\u003eFood Hydrocolloids\u003c/em\u003e \u003cstrong\u003e63\u003c/strong\u003e, 170-177 (2017). \u003c/li\u003e\n\u003cli\u003eJuanjuan, D.\u003cem\u003e et al.\u003c/em\u003e The effect of fibrin on rheological behavior, gelling properties and microstructure of myofibrillar proteins. \u003cem\u003eLWT\u003c/em\u003e \u003cstrong\u003e153\u003c/strong\u003e (2022). \u003c/li\u003e\n\u003cli\u003eHongxia, G.\u003cem\u003e et al.\u003c/em\u003e Review of recent advances in the preparation, properties, and applications of high internal phase emulsions. \u003cem\u003eTrends in Food Science \u0026amp; Technology\u003c/em\u003e\u003cstrong\u003e112\u003c/strong\u003e, 36-49 (2021).\u003c/li\u003e\n\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":true,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"npj-science-of-food","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"npjscifood","sideBox":"Learn more about [npj Science of Food](http://www.nature.com/npjscifood/)","snPcode":"41538","submissionUrl":"https://submission.springernature.com/new-submission/41538/3","title":"npj Science of Food","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"stoa","reportingPortfolio":"NPJ","inReviewEnabled":true,"inReviewRevisionsEnabled":true},"keywords":"Rice Bran Protein, Protein Fiber Aggregates, Mayonnaise-like Products, Rheological Properties","lastPublishedDoi":"10.21203/rs.3.rs-7678095/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-7678095/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eIt was investigated the impact of rice bran protein (RBP) fiber (PF) aggregates on the properties of mayonnaise-like emulsions. Using RBP with different structures during PF aggregate formation, plant-based emulsions were prepared and evaluated for interfacial protein adsorption (AP%), water-holding capacity (WHC), rheological and textural properties, color, sensory quality, and storage stability. PF aggregate\u0026ndash;based emulsions had 11.29% and 16.8% higher AP% values and 43.73% and 107.64% higher WHC values than those of RBP emulsions and homemade mayonnaise, respectively. Texture analysis showed significant improvements in hardness, viscosity, cohesiveness, and springiness. After 30 days, peroxide values in PF aggregate emulsions were 25.13% lower than those in RBP emulsions and 33.57% lower than those in homemade mayonnaise; malondialdehyde content was 15.57% and 21.25% lower, respectively. The total viable count of PF aggregate emulsions was 21.51% lower than that of RBP emulsions. These findings highlight the enhanced stability of PF aggregate\u0026ndash;based mayonnaise-like emulsions.\u003c/p\u003e","manuscriptTitle":"Edible mayonnaise-like emulsion stabilized by rice bran protein fibril aggregation: Effect of fibril aggregate structure","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-11-20 12:47:34","doi":"10.21203/rs.3.rs-7678095/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Revision requested","date":"2025-11-15T13:16:39+00:00","index":"","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2025-11-15T06:10:54+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"40399776072014771089406143053778659470","date":"2025-11-13T07:47:24+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"174832066931014187052972665631004762384","date":"2025-11-13T07:46:34+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2025-11-12T07:48:09+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"295174254629139682816015202248049433987","date":"2025-11-11T15:05:38+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"191549796781365702190930283228419067524","date":"2025-11-11T11:53:26+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"12892732747381921399797531945534555125","date":"2025-11-11T07:07:37+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"83765749979637295887022480691261260746","date":"2025-11-11T06:35:02+00:00","index":"hide","fulltext":""},{"type":"reviewersInvited","content":"","date":"2025-11-11T06:13:16+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2025-10-15T13:08:09+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2025-10-08T06:11:33+00:00","index":"","fulltext":""},{"type":"submitted","content":"npj Science of Food","date":"2025-09-22T10:34:15+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"npj-science-of-food","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"npjscifood","sideBox":"Learn more about [npj Science of Food](http://www.nature.com/npjscifood/)","snPcode":"41538","submissionUrl":"https://submission.springernature.com/new-submission/41538/3","title":"npj Science of Food","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"stoa","reportingPortfolio":"NPJ","inReviewEnabled":true,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"5ee8a415-dfae-4794-878b-51c0e5a9b6f1","owner":[],"postedDate":"November 20th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"published-in-journal","subjectAreas":[{"id":58044284,"name":"Physical sciences/Chemistry"},{"id":58044285,"name":"Physical sciences/Engineering"},{"id":58044286,"name":"Physical sciences/Materials science"}],"tags":[],"updatedAt":"2026-01-19T16:45:29+00:00","versionOfRecord":{"articleIdentity":"rs-7678095","link":"https://doi.org/10.1038/s41538-025-00689-0","journal":{"identity":"npj-science-of-food","isVorOnly":false,"title":"npj Science of Food"},"publishedOn":"2026-01-12 16:28:47","publishedOnDateReadable":"January 12th, 2026"},"versionCreatedAt":"2025-11-20 12:47:34","video":"","vorDoi":"10.1038/s41538-025-00689-0","vorDoiUrl":"https://doi.org/10.1038/s41538-025-00689-0","workflowStages":[]},"version":"v1","identity":"rs-7678095","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-7678095","identity":"rs-7678095","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}