Exploring community succession and metabolic changes in corn gluten meal-bran mixed wastes during fermentation.

Addressing the challenge of sustainable agricultural processing waste management is crucial. Protein sources are essential for livestock farming, and one viable solution is the microbial fermentation of agricultural by-products. In this study, the microorganisms utilized for fermentation were Pichia fermentans PFZS and Limmosilactobacillus fermentum LFZS. The results demonstrated that the fermented corn gluten meal-bran mixture (FCBM) effectively degraded high molecular weight proteins, resulting in increases of approximately 23.3%, 367.6%, and 159.3% in crude protein (CP), trichloroacetic acid-soluble protein (TCA-SP), and free amino acid (FAA), respectively. Additionally, there was a significant enhancement in the content of beneficial metabolites, including total phenols, carotenoids, and microorganisms. FCBM also effectively reduced anti-nutritional factors while boosting antioxidant and anti-inflammatory substances, such as dipeptides and tripeptides. The fermentation process was marked by an increase in beneficial endophytes, which was closely correlated with the enhancement of beneficial metabolites. Overall, FCBM provides a theoretical basis for substituting traditional protein resources in animal husbandry.

Silencing circular RNA hsa_circABCC1 inhibits osteosarcoma progression through down-regulating HDAC4 via sponging miR-591.

BACKGROUND: Circular RNA (circRNA) has been shown to play an important regulatory role in the development of various cancers, including osteosarcoma (OS). However, the role of circRNA ABCC1 (circABCC1) in OS was still poorly understood. The aim of our study was to investigate the role of circABCC1 in OS progression and its potential molecular mechanisms. METHODS: The expression of circABCC1, microRNA-591 (miR-591) and histone deacetylase 4 (HDAC4) in OS tissues or cells was detected by quantitative real-time polymerase chain reaction (qRT-PCR) and western blot (WB) analyses. In vitro experiments, the viability, proliferation, apoptosis, migration, invasion and autophagy of U2OS and HOS cells were assessed in vitro using cell counting kit-8 (CCK-8) assay, 5-ethynyl-29-deoxyuridine (EdU) assay, flow cytometry (FCM) assay, transwell migration and invasion assays (transwell) and WB assay, respectively. Interactions between circABCC1 and miR-591, miR-591 and HDAC4 were confirmed using a dual luciferase reporter gene assay system. The oncogenic role of circABCC1 in OS in vivo was examined by establishing a tumor xenograft model. RESULTS: CircABCC1 was significantly elevated in OS tissues (about 3.1-folds) and cells (U2OS (about 2.1-folds) and HOS (about 2.8-folds)) compared with the control (p < .05). Silencing of circABCC1 significantly reduced the viability and proliferation, promoted apoptosis, impaired migration and invasion, and increased autophagy of U2OS and HOS cells (p < .05). In addition, miR-591 was confirmed to be a target of circABCC1, exerting an opposite effect to circABCC1 (p < .05). MiR-591 attenuation in U2OS and HOS cells was able to reply to the inhibition of cell proliferation, migration and invasion as well as promotion of cell apoptosis and autophagy mediated by silencing circABCC1 (p < .05). HDAC4 was verified to be the target gene of miR-591 in U2OS and HOS cells and was regulated by the circABCC1/miR-591 axis (p < .05), and restoration of HDAC4 levels in U2OS and HOS cells was able to restore the altered cellular function caused by silencing circABCC1 (p < .05). In addition, knockdown of circABCC1 attenuated tumor growth in vivo (p < .05). CONCLUSION: Silencing of circABCC1 inhibits osteosarcoma progression by attenuating HDAC4 expression through sponging miR-591.

Metabolite analysis of soybean oil on promoting astaxanthin production of Phaffia rhodozyma.

BACKGROUND: Astaxanthin is a carotenoid with strong antioxidant property. In addition, it has anti-cancer, anti-tumor, anti-inflammatory and many other functions. The micro-organisms that mainly produce astaxanthin are Haematococcus pluvialis and Phaffia rhodozyma. Compared with H. pluvialis, P. rhodozyma has shorter fermentation cycle and easier to control culture conditions, but the yield of astaxanthin in P. rhodozyma is low. This article studied how to improve the astaxanthin production of P. rhodozyma. RESULTS: The results showed that when 10 mL L-1 soybean oil was added to the culture medium, astaxanthin production increased significantly, reaching 7.35 mg L-1 , which was 1.4 times that of the control group, and lycopene and ß-carotene contents also increased significantly. Through targeted metabolite analysis, the fatty acids in P. rhodozyma significantly increased under the soybean oil stimulation, especially the fatty acids closely related to the formation of astaxanthin esters, included palmitic acid (C16:0), stearic acid (C18:0), oleic acid (C18:1n9), linoleic acid (C18:2n6), α-linolenic acid (C18:3n3) and γ-linolenic acid (C18:3n6), thereby increasing the astaxanthin esters content. CONCLUSION: It showed that the addition of soybean oil can promote the accumulation of astaxanthin by promoting the increase of astaxanthin ester content. © 2022 Society of Chemical Industry.

The ameliorative effect of probiotics on diet-induced lipid metabolism disorders: A review.

High-fat diet induces lipid metabolism disorders that has become one of the grievous public health problems and imposes a serious economic and social burden worldwide. Safety probiotics isolated from nature are regarded as a novel supplementary strategy for preventing and improving diet-induced lipid metabolism disorders and related chronic diseases. The present review summarized the latest researches of probiotics in high fat diet induced lipid metabolism disorders to provide a critical perspective on the regulatory function of probiotics for future research. Furthermore, the screening criteria and general sources of probiotics with lipid-lowering ability also outlined to enlarge microbial species resource bank instantly, which promoted the development of functional foods with lipid-lowering strains from nature. After critically reviewing the lipid-lowering potential of probiotics both in vitro and in vivo and even in clinical data of humans, we provided a perspective that probiotics activated AMPK signaling pathway to regulate fat synthesis and decomposition, as well as affected positively the gut microbiota structure, intestinal barrier function and systemic inflammatory response, then these beneficial effects are amplified along Gut-liver axis, which regulated intestinal flora metabolites such as SCFAs and BAs by HMGCR/FXR/SHP signaling pathway to improve high fat diet induced lipid metabolism disorders effectively.

Effects of Ultra-High Pressure on Endogenous Enzyme Activities, Protein Properties, and Quality Characteristics of Shrimp (Litopenaeus vannamei) during Iced Storage.

The present study aimed to explore the effects of ultra-high pressure (UHP) on the cathepsin (B, D, H, and L) activities, protein oxidation, and degradation properties as well as quality characteristics of iced shrimp (Litopenaeus vannamei). Fresh shrimps were vacuum-packed, treated with UHP (100-500 MPa for 5 min), and stored at 0 °C for 15 days. The results showed that the L* (luminance), b* (yellowness), W (whiteness), ΔE (color difference), hardness, shear force, gumminess, chewiness, and resilience of shrimp were significantly improved by UHP treatment. Moreover, the contents of surface hydrophobicity, myofibril fragmentation index (MFI), trichloroacetic acid (TCA)-soluble peptides, carbonyl, dityrosine, and free sulfhydryl of myofibrillar protein (MP) were significantly promoted by UHP treatment. In addition, UHP (above 300 MPa) treatment enhanced the mitochondrial membrane permeability but inhibited the lysosomal membrane stability, and the cathepsin (B, D, H, and L) activities. UHP treatment notably inhibited the activities of cathepsins, delayed protein oxidation and degradation, as well as texture softening of shrimp during storage. Generally, UHP treatment at 300 MPa for 5 min effectively delayed the protein and quality deterioration caused by endogenous enzymes and prolonged the shelf life of shrimp by 8 days.

Effect of pullulan concentration and pH on the interactions between whey protein concentrate and pullulan during gelation.

BACKGROUND: Whey protein concentrate (WPC)/pullulan (PUL) hydrogel is applied as a microencapsulation wall material to protect probiotics. However, the interactions between WPC and PUL during gelation have not been clarified. In the present study, the effects of PUL concentration and pH on the interactions between WPC and PUL during gelation were evaluated with respect to appearance, zeta-potential, sulfhydryl group amount, surface hydrophobicity and infrared spectroscopy measurements. The rheological properties of WPC/PUL gels were also determined. RESULTS: The results obtained showed that a proper concentration (0.40 g mL-1 ) of PUL could improve the gel by enhancing the strength of hydrogen bonding, electrostatic interactions and exposure of hydrophobic groups, whereas too much PUL inhibited the formation of disulfide bonds. Furthermore, hydrophobic interactions, disulfide bonds and hydrogen bonds were destroyed in varying degrees under an alkaline environment. The rheological results also demonstrated a similar effect of PUL concentration and pH on the storage modulus (G') of WPC/PUL gels. CONCLUSION: When the WPC/PUL gel was formed at PUL concentration of 0.40 g mL-1 and pH 7.0, the interaction between WPC and PUL could be enhanced, which is beneficial for the future application of WPC/PUL gels in the food industry. © 2020 Society of Chemical Industry.

Enzymatic transformation of ginsenosides Re, Rg1, and Rf to ginsenosides Rg2 and aglycon PPT by using ß-glucosidase from Thermotoga neapolitana.

OBJECTIVES: To enzymatically transform protopanaxatriol by using ß-glucosidase from Thermotoga neapolitana (T. neapolitana) DSM 4359. RESULTS: Recombinant ß-glucosidase was purified, which molecular weight was about 79.5 kDa. High levels of ginsenoside were obtained using the follow reaction conditions: 2 mg ml-1 ginsenoside, 25 U ml-1 enzyme, 85 °C, and pH 5.0. ß-glucosidase converted ginsenoside Re to Rg2, Rf and Rg1 to APPT completely after 3 h under the given conditions, respectively. The enzyme created 1.66 mg ml-1 Rg2 from Re with 553 mg l-1 h-1, 0.85 mg ml-1, and 1.01 mg ml-1 APPT from Rg1 and Rf with 283 and 316 mg l-1 h-1 APPT. CONCLUSIONS: ß-glucosidase could be useful for the high-yield, rapid, and low-cost preparation of ginsenoside Rg2 from Re, and APPT from the ginsenosides Rg1 and Rf.

Comparative Transcriptome Analyses Provide Potential Insights into the Molecular Mechanisms of Astaxanthin in the Protection against Alcoholic Liver Disease in Mice.

Alcoholic liver disease (ALD) is a major cause of chronic liver disease worldwide. It is a complex process, including a broad spectrum of hepatic lesions from fibrosis to cirrhosis. Our previous study suggested that astaxanthin (AST) could alleviate the hepatic inflammation and lipid dysmetabolism induced by ethanol administration. In this study, a total of 48 male C57BL/6J mice were divided into 4 groups: a Con group (fed with a Lieber⁻DeCarli liquid diet), an AST group (fed with a Lieber⁻DeCarli liquid diet and AST), an Et group (fed with an ethanol-containing Lieber⁻DeCarli liquid diet), and a EtAST group (fed with an ethanol-containing Lieber⁻DeCarli liquid diet and AST). Then, comparative hepatic transcriptome analysis among the groups was performed by Illumina RNA sequencing. Gene enrichment analysis was conducted to identify pathways affected by the differentially expressed genes. Changes of the top genes were verified by quantitative real-time PCR (qRT-PCR) and Western blot. A total of 514.95 ± 6.89, 546.02 ± 15.93, 576.06 ± 21.01, and 690.85 ± 54.14 million clean reads were obtained for the Con, AST, Et, and EtAST groups, respectively. Compared with the Et group, 1892 differentially expressed genes (DEGs) (including 351 upregulated and 1541 downregulated genes) were identified in the AST group, 1724 differentially expressed genes (including 233 upregulated and 1491 downregulated genes) were identified in the Con group, and 1718 DEGs (including 1380 upregulated and 338 downregulated genes) were identified in the EtAST group. The enrichment analyses revealed that the chemokine signaling, the antigen processing and presentation, the nucleotide-binding and oligomerization domain (NOD)-like receptor signaling, and the Toll-like receptor signaling pathways enriched the most differentially expressed genes. The findings of this study provide insights for the development of nutrition-related therapeutics for ALD.

Anterior versus posterior instrumentation for treatment of thoracolumbar tuberculosis : A meta-analysis.

BACKGROUND: To compare clinical and functional outcomes of anterior versus posterior debridement and spinal fixation for surgical treatment of thoracolumbar tuberculosis. METHODS: A computer-based online search of the Cochrane Library, PubMed, EMBase, Wanfang, VIP, and the CNKI database was performed. The methodological quality of included studies was evaluated, and data analyses were performed using RevMan 5.0 software (The Nordic Cochrane Centre, The Cochrane Collaboration Copenhagen, Denmark). RESULTS: Eleven trials were studied, with eight performed in China, two in Egypt, and one in India. The results showed significant differences between the two operative approaches in terms of correction of kyphotic angle and intraoperative blood loss, but not in terms of operation time, hospital stay, fusion time, and loss of correction at the final follow-up. CONCLUSION: The anterior and posterior approaches are equally good methods for treatment of thoracolumbar tuberculosis. The anterior approach results in less blood loss, whereas posterior instrumentation is better suited for correction of kyphotic angle.

Effect of fermentation on water mobility and distribution in fermented cornmeal using LF-NMR and its correlation with substrate.

Cordyceps militaris (C. militaris) was utilized to ferment cornmeal by solid state fermentation. The main objective of this study was to investigate effect of fermentation on the dynamic state of water and microstructure distribution of water within cornmeal with Low-field nuclear magnetic resonance, as well as the effect on composition and microstructure properties. The spin-spin relaxation time (T2) showed significant changes in solid-state fermented cornmeal. Principal component analysis further revealed that the variations within different fermentation stage could be discriminated by the T2 parameters. Bulk water (T22) was the main form of water present and lost in substrates. The weights of different indicators, as assessed by multiple regression analysis, demonstrated that there was a strong correlation between starch and T2 relaxation. Scanning electron microscopy demonstrated that fermentation can cause the appearance of micropores. The longer relaxation time of T22 during logarithmic period can be interpreted as a loosening of the structure at the starch hydrolysis, introducing more water into the structure. Thus, the differences in composition and structure of the substrate at different fermentation time produce different T2 values.

Determining electrospun morphology from the properties of protein-polymer solutions.

Integrating natural macromolecules, e.g. proteins, is a progressive trend in the fabrication of biocompatible sub-micrometer fibers with tunable diameters using the electrospinning technique. The correlation between solution properties and electrospun morphology is critical; it is quite clear for synthetic linear polymer solutions but remains uncertain for solutions with protein. Here, we report the determination of electrospun morphology in protein-polymer solutions of poly(ethylene oxide) (PEO) and zein, a storage protein from corn. The viscosity of the zein/PEO mixed solutions can be well described using the Lederer-Roegiers equation and decreases with the increase of the fraction of zein. The surface tension sharply decreases above a critical concentration at the saturation of the interfacial monolayer. Correspondingly, the different electrospun morphologies-from bead, coexisting bead and fiber, to fiber and ribbon-were mapped onto a ternary phase diagram and a viscosity contour plot. Such coupling provides a clear way to determine the electrospun morphology from solution properties. The occurrence of electrospun fibers partially follows two empirical rules, while the critical point revealed from surface tension has the best approximation. The diameters of electrospun fibers were found to have a scaling relationship against concentration, zero-shear viscosity and surface tension of solutions. These scaling exponents were compared with those from typical polymer solutions. The analysis suggests that aqueous ethanol gives different solvent qualities to zein and PEO solutions, resulting in the irregular shape in the phase diagram that correlates solution properties and electrospun morphologies.

Preparation of Octadecylamine-Graphene/MgCl2-Supported Ziegler-Natta Catalyst and Its Application in Ethylene Polymerization.

A facile coagglomeration method for preparing a long alkyl chain modified graphene oxide (MGO)/MgCl2-supported Ti-based Ziegler-Natta catalyst was reported. The effects of MGO on the catalyst morphology and activity for ethylene polymerization were examined. The resultant polyethylene (PE)/MGO nanocomposites exhibited a layered morphology, with the MGO fillers being well dispersed and exhibiting strong interfacial adhesion to the PE matrix. The thermal stability and mechanical properties of the PE were significantly enhanced with the introduction of a small amount of the MGO filler. Thus, this work provides a facile approach to the production of high-performance PE.

Antioxidant hydrolyzed peptides from Manchurian walnut (Juglans mandshurica Maxim.) attenuate scopolamine-induced memory impairment in mice.

BACKGROUND: Walnut protein, which is obtained as a by-product of oil expression, has not been used efficiently. Although walnuts are beneficial for cognitive functioning, the potential of their protein composition in strengthening learning and memory functions remains unknown. In this study, the inhibition of memory impairment by the Manchurian walnut hydrolyzed peptide (MWHP) was evaluated. RESULTS: Small-molecular-weight MWHP (<3 kDa) achieved the optimal antioxidative activity. Therefore, MWHP (<3 kDa) was subjected to the following mice trials to evaluate its attenuation effect on memory impairment. In the Morris water maze test, MWHP shortened the total path for searching the platform, reduced the escape latency, and increased the dwelling distance and time in the coverage zone. MWHP also prolonged the latency and diminished errors in the passive avoidance response tests. These behavioral tests demonstrated that MWHP could inhibit scopolamine-induced memory impairment. MWHP improved memory by reducing oxidative stress, inhibiting apoptosis, regulating neurotransmitter functions, maintaining hippocampal CA3 pyramidal neurons, and increasing calmodulin-dependent protein kinase II levels in brain tissues. CONCLUSION: Experimental results proved that MWHP exhibits potential in improving memory and should be used to develop novel functional food. © 2018 Society of Chemical Industry.

Synthesis and Characterization of Polyethylene/Starch Nanocomposites: A Spherical Starch-Supported Catalyst and In Situ Ethylene Polymerization.

In the present article, a novel spherical starch-supported vanadium (V)-based Ziegler-Natta catalyst was synthesized. The active centers of the obtained catalyst well dispersed in the starch through the SEM-EDX analysis. The effects of reaction conditions on ethylene polymerization were studied. The synthesized catalyst exhibited high activity toward ethylene polymerization in the presence of ethylaluminium sesquichloride (EASC) cocatalyst. Interestingly, the fiber shape PE was obtained directly during the polymerization process.

Gastrodin Attenuates Cognitive Deficits Induced by 3,3'-Iminodipropionitrile.

3,3'-Iminodipropionitrile (IDPN), one of the nitrile derivatives, can induce persistent neurotoxicity, and therefore cause dyskinesia and cognitive impairments. Gastrodin, a main bioactive ingredient of Gastrodia elata Blume, is shown to greatly improve cognitive function. The aim of this study was to further determine whether administration of gastrodin can ameliorate IDPN-induced cognitive deficits in the Morris water maze (MWM) and novel object recognition (NOR) task, and to explore the underlying mechanisms. Results showed that exposure to IDPN (100 mg/kg/day, for 8 days) significantly impaired spatial and object recognition memory and that repeated treatment with gastrodin (150 mg/kg/day, for 6 weeks) could effectively alleviate the IDPN-induced cognitive impairments as indicated by increased spatial memory and discrimination ratio in the MWM and NOR tests. Gastrodin treatment also reverted IDPN-induced decreases of γ-aminobutyric acid (GABA) levels and increases of a2 GABAA receptor protein expression in the prefrontal cortex and hippocampus of IDPN-treated rats. These results suggest that gastrodin treatment may provide a novel pharmacological strategy for IDPN-induced cognitive deficits, which was mediated, at least in part, by normalizing the GABAergic system.

In vivo assessment of immunomodulatory activity of hydrolysed peptides from Corylus heterophylla Fisch.

BACKGROUND: Hazelnut dregs are by-products of hazelnut oil expression, which have not been fully exploited. This research aims to assess the immunomodulatory function of hazelnut hydrolysed peptides (HHPs). RESULTS: HHPs with a hydrolysis degree of 38.08% were divided into three fractions by ultra-filtration: the high molecular weight peptide (>10 kDa), medium molecular weight peptide (3 kDa to 10 kDa), and low molecular weight peptide (<3 kDa). Mice were fed daily with HHPs of different molecular weights at doses of 200, 400, and 800 mg kg(-1) body weight. On the 10th, 20th and 30th day of feeding, representative immune indexes were measured. Results showed that HHPs can regulate the immune system of mice, which is affected by the molecular weight of HHP and the feeding time. Generally, short-term feeding (10 d to 20 d) with HHPs of different molecular weights can improve most immune indexes (organ index, spleen lymphocyte proliferation, macrophage activity, secretory immunoglobulin A content, and number of CD4(+) and CD8(+) T cells), whereas during long-term feeding (30 d), low molecular weight HHP can better sustain immune regulation. CONCLUSION: HHPs exhibit potential immunomodulatory properties, which has promising implications for the development of new functional foods. © 2015 Society of Chemical Industry.

Characterization of Aspartate Kinase from Corynebacterium pekinense and the Critical Site of Arg169.

Aspartate kinase (AK) is the key enzyme in the biosynthesis of aspartate-derived amino acids. Recombinant AK was efficiently purified and systematically characterized through analysis under optimal conditions combined with steady-state kinetics study. Homogeneous AK was predicted as a decamer with a molecular weight of ~48 kDa and a half-life of 4.5 h. The enzymatic activity was enhanced by ethanol and Ni(2+). Moreover, steady-state kinetic study confirmed that AK is an allosteric enzyme, and its activity was inhibited by allosteric inhibitors, such as Lys, Met, and Thr. Theoretical results indicated the binding mode of AK and showed that Arg169 is an important residue in substrate binding, catalytic domain, and inhibitor binding. The values of the kinetic parameter Vmax of R169 mutants, namely, R169Y, R169P, R169D, and R169H AK, with l-aspartate as the substrate, were 4.71-, 2.25-, 2.57-, and 2.13-fold higher, respectively, than that of the wild-type AK. Furthermore, experimental and theoretical data showed that Arg169 formed a hydrogen bond with Glu92, which functions as the entrance gate. This study provides a basis to develop new enzymes and elucidate the corresponding amino acid production.

Computational study on substrate specificity of a novel cysteine protease 1 precursor from Zea mays.

Cysteine protease 1 precursor from Zea mays (zmCP1) is classified as a member of the C1A family of peptidases (papain-like cysteine protease) in MEROPS (the Peptidase Database). The 3D structure and substrate specificity of the zmCP1 is still unknown. This study is the first one to build the 3D structure of zmCP1 by computer-assisted homology modeling. In order to determine the substrate specificity of zmCP1, docking study is used for rapid and convenient analysis of large populations of ligand-enzyme complexes. Docking results show that zmCP1 has preference for P1 position and P2 position for Arg and a large hydrophobic residue (such as Phe). Gly147, Gly191, Cys189, and Asp190 are predicted to function as active residues at the S1 subsite, and the S2 subsite contains Leu283, Leu193, Ala259, Met194, and Ala286. SIFt results indicate that Gly144, Arg268, Trp308, and Ser311 play important roles in substrate binding. Then Molecular Mechanics-Poisson-Boltzmann Surface Area (MM-PBSA) method was used to explain the substrate specificity for P1 position of zmCp1. This study provides insights into the molecular basis of zmCP1 activity and substrate specificity.

Research progress of protein complex systems and their application in food: A review.

Among the common natural biomolecules, the excellent properties of proteins have attracted extensive attention from researchers for functional applications, however, in native form proteins have many limitations in the performance of their functional attribute. However, with the deepening of research, it has been found that the combination of natural active substances such as polyphenols, polysaccharides, etc. with protein molecules will make the composite system have stronger functional properties, while the utilization of pH-driven method, ultrasonic treatment, heat treatment, etc. not only provides a guarantee for the overall protein-based composite system, but also gives more possibilities to the protein-composite system. Protein composite systems are emerging in the fields of novel active packaging, functional factor delivery systems and gel systems with high medical value. The products of these protein composite systems usually have high functional properties, mainly due to the interaction of the remaining natural active substances with protein molecules, which can be broadly categorized into covalent interactions and non-covalent interactions, and which, despite the differences in these interactions, together constitute the cornerstone for the stability of protein composite systems and for in-depth research.

Research on corn starch and black bean protein isolate interactions during gelatinization and their effects on physicochemical properties of the blends.

The interaction between starch and protein during food processing is crucial for controlling food quality. This study aims to understand the interactions between corn starch and black bean protein isolate (BBPI) at various gelatinization phases and their effects on the physicochemical properties of the blends. BBPI reduced the rheological properties of the corn starch/BBPI mixed system during gelatinization, increasing light transmittance and gelatinization temperature, while decreasing total viscosity and enthalpy change. The changes in starch and protein microstructure during gelatinization indicated that BBPI adhered to the starch particle surface or partially penetrated the swollen starch particles. Fourier transform infrared spectroscopy (FT-IR) revealed that BBPI decreased the number of hydrogen bonds within starch, with no newly formed functional groups in the mixed system. Furthermore, BBPI reduced the composite relative crystallinity (RC). The effect of protein addition on water migration in the mixed system demonstrates that protein and starch compete for water during gelatinization, preventing water molecules from diffusing into starch particles.