Synergistic effects of tilapia head protein hydrolysate and walnut protein hydrolysate on the amelioration of cognitive impairment in mice.

BACKGROUND: Cognitive impairment (CI) is a significant public health concern, and bioactive peptides have shown potential as therapeutic agents. However, information about their synergistic effects on cognitive function is still limited. Here, we investigated the synergistic effects of tilapia head protein hydrolysate (THPH) and walnut protein hydrolysate (WPH) in mitigating CI induced by scopolamine in mice. RESULTS: The results showed that the combined supplementation of THPH and WPH (mass ratio, 1:1) was superior to either individual supplement in enhancing spatial memory and object recognition abilities in CI mice, and significantly lessened brain injury in CI mice by alleviating neuronal damage, reducing oxidative stress and stabilizing the cholinergic system. In addition, the combined supplementation was found to be more conducive to remodeling the gut microbiota structure in CI mice by not only remarkably reducing the ratio of Firmicutes to Bacteroidota, but also specifically enriching the genus Roseburia. On the other hand, the combined supplementation regulated the disorders of sphingolipid and amino acid metabolism in CI mice, particularly upregulating glutathione and histidine metabolism, and displayed a stronger ability to increase the expression of genes and proteins related to the brain-derived neurotrophic factor (BDNF)/TrkB/CrEB signaling pathway in the brain. CONCLUSION: These findings demonstrate that tilapia head and walnut-derived protein hydrolysates exerted synergistic effects in ameliorating CI, which was achieved through modulation of gut microbiota, serum metabolic pathways and BDNF signaling pathways. © 2024 Society of Chemical Industry.

A physicochemical double-cross-linked gelatin hydrogel with enhanced antibacterial and anti-inflammatory capabilities for improving wound healing.

BACKGROUND: Skin tissue is vital in protecting the body from injuries and bacterial infections. Wound infection caused by bacterial colonization is one of the main factors hindering wound healing. Wound infection caused by colonization of a large number of bacteria can cause the wound to enter a continuous stage of inflammation, which delays wound healing. Hydrogel wound dressing is composed of natural and synthetic polymers, which can absorb tissue fluid, improve the local microenvironment of wound, and promote wound healing. However, in the preparation process of hydrogel, the complex preparation process and poor biological efficacy limit the application of hydrogel wound dressing in complex wound environment. Therefore, it is particularly important to develop and prepare hydrogel dressings with simple technology, good physical properties and biological effects by using natural polymers. RESULTS: In this study, a gelatin-based (Tsg-THA&Fe) hydrogel was created by mixing trivalent iron (Fe3+) and 2,3,4-trihydroxybenzaldehyde (THA) to form a complex (THA&Fe), followed by a simple Schiff base reaction with tilapia skin gelatin (Tsg). The gel time and rheological properties of the hydrogels were adjusted by controlling the number of complexes. The dynamic cross-linking of the coordination bonds (o-phthalmictriol-Fe3+) and Schiff base bonds allows hydrogels to have good self-healing and injectable properties. In vitro experiments confirmed that the hydrogel had good biocompatibility and biodegradability as well as adhesion, hemostasis, and antibacterial properties. The feasibility of Tsg-THA&Fe hydrogel was studied by treating rat skin trauma model. The results showed that compared with Comfeel® Plus Transparent dressing, the Tsg-THA&Fe hydrogel could obvious reduce the number of microorganisms, prevent bacterial colonization, reduce inflammation and accelerate wound healing. Local distribution of the Tsg-THA&Fe hydrogel in the skin tissue did not cause organ toxicity. CONCLUSIONS: In summary, the preparation process of Tsg-THA&Fe hydrogel is simple, with excellent performance in physical properties and biological efficacy. It can effectively relieve inflammation and control the colonization of wound microbes, and can be used as a multi-functional dressing to improve wound healing.

Comparative Study on Curcumin Loaded in Golden Pompano (Trachinotus blochii) Head Phospholipid and Soybean Lecithin Liposomes: Preparation, Characteristics and Anti-Inflammatory Properties.

In this study, we compared the characteristics and in vitro anti-inflammatory effects of two curcumin liposomes, prepared with golden pompano head phospholipids (GPL) and soybean lecithin (SPC). GPL liposomes (GPL-lipo) and SPC liposomes (SPC-lipo) loaded with curcumin (CUR) were prepared by thin film extrusion, and the differences in particle size, ζ-potential, morphology, and storage stability were investigated. The results show that GPL-lipo and SPC-lipo were monolayer liposomes with a relatively small particle size and excellent encapsulation rates. However, GPL-lipo displayed a larger negative ζ-potential and better storage stability compared to SPC-lipo. Subsequently, the effects of phospholipids in regulating the inflammatory response of macrophages were evaluated in vitro, based on the synergistic effect with CUR. The results showed that both GPL and SPC exerted excellent synergistic effect with CUR in inhibiting the lipopolysaccharide (LPS)-induced secretion of nitric oxide (NO), reactive oxygen species (ROS), and pro-inflammatory genes (tumor necrosis factor (TNF)-α, interleukin 1ß (IL-ß), and interleukin 6 (IL-6)) in RAW264.7 cells. Interestingly, GPL-lipo displayed superior inhibitory effects, compared to SPC-lipo. The findings provide a new innovative bioactive carrier for development of stable CUR liposomes with good functional properties.

Effect of citric acid on physicochemical properties and protein structure of low-salt restructured tilapia (Oreochromis mossambicus) meat products.

BACKGROUND: The growing consumer demand for healthy products has encouraged the development of low-salt meat products. In this study, to develop low-salt restructured tilapia (Oreochromis mossambicus) meat products, citric acid was used to improve the properties of restructured tilapia products. RESULTS: In comparison with control restructured fish products (RP) and surimi products (SP), 0.2% citric acid-treated restructured fish products (RPC) and surimi products (SPC) showed a significant decrease in expressible water and water activity and a remarkable increase in whiteness, dry matter, hardness, chewiness, gumminess, and acceptability. Mechanistic studies suggested that citric acid significantly changed the content of total protein and myofibrillar proteins and promoted degradation of heavy myosin chains. Fourier-transform infrared and Raman spectra revealed the citric acid-mediated alteration in the peak intensities of amide I and amide II bands, which changed the secondary structures of RPC and SPC. CONCLUSION: It is feasible to prepare low-salt restructured tilapia meat products using citric acid, which offers a means of using muscle by-products and exploiting new functional products with an added commercial value. © 2020 Society of Chemical Industry.

Holothuria Leucospilota Polysaccharides Ameliorate Hyperlipidemia in High-Fat Diet-Induced Rats via Short-Chain Fatty Acids Production and Lipid Metabolism Regulation.

Holothuria leucospilota polysaccharides (HLP) are expected to become potential resources for the treatment of hyperlipidemia because of their various bioactivities. In the study, the treatment of HLP on improving hyperlipidemia in rats was explored. Oral administration of HLP at 100 or 200 mg/kg body weight effectively alleviated serum lipid levels and liver histological abnormalities in high-fat-diet rats. HLP regulated abnormal mRNA, lipogenesis-related hormones and inflammatory cytokines (tumor necrosis factor-α, interleukin-6 and interleukin-12) levels. HLP improved the ability of gut microbiota to produce short-chain fatty acids (SCFAs). SCFAs have been found to ameliorate liver lesions. Therefore, HLP alleviated hyperlipidemia by improving the levels of SCFAs to regulate lipid metabolism. These results indicated that HLP could be used as beneficial polysaccharides to alleviate hyperlipidemia.

Isolation and Characterization of a Novel Sialoglycopeptide Promoting Osteogenesis from Gadus morhua Eggs.

Gadus morhua eggs contain several nutrients, including polyunsaturated fatty acids, lecithin and glycoproteins. A novel sialoglycopeptide from the eggs of G. morhua (Gm-SGPP) was extracted with 90% phenol and purified by Q Sepharose Fast Flow (QFF) ion exchange chromatography, followed by S-300 gel filtration chromatography. Gm-SGPP contained 63.7% carbohydrate, 16.2% protein and 18.6% N-acetylneuraminic acid. High-performance size exclusion chromatography and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) demonstrated that Gm-SGPP is a 7000-Da pure sialoglycopeptide. ß-elimination reaction suggested that Gm-SGPP contained N-glycan units. Amino acid N-terminal sequence analysis indicated the presence of Ala-Ser-Asn-Gly-Thr-Gln-Ala-Pro amino acid sequence. Moreover, N-glycan was connected at the third Asn location of the peptide chain through GlcNAc. Gm-SGPP was composed of D-mannose, D-glucuronic acid and D-galactose. Fourier transform-infrared spectroscopy (FT-IR), 1H-nuclear magnetic resonance spectroscopy (1H-NMR) and methylation analysis were performed to reveal the structure profile of Gm-SGPP. In vitro results showed that the proliferation activity of MC3T3-E1 cells was significantly promoted by Gm-SGPP. In vivo data revealed that Gm-SGPP increased the calcium and phosphorus content of tibias and promoted longitudinal bone growth in adolescent rats.

Multi-targeting inulin-based nanoparticles with cannabidiol for effective prevention of ulcerative colitis.

The pathogenesis of ulcerative colitis (UC) is closely related to severe inflammation, damaged colonic mucosal barrier, increased oxidative stress and intestinal ecological imbalance. However, due to the nonspecific distribution and poor bioavailability of drugs, UC treatment is still a serious challenge. Here, a mitochondria/colon dual targeted nanoparticles based on redox response was developed to effectively alleviate UC. Cannabidiol nanoparticles (CBD NPs) with a particle size of 143.2 ± 3.11 nm were prepared by self-assembly using polymers (TPP-IN-LA) obtained by modifying inulin with (5-carboxypentyl) triphenyl phosphonium bromide (TPP) and α-lipoic acid (α-LA). Excitingly, the constructed CBD NPs showed excellent mitochondrial targeting, with a Pearson correlation coefficient of 0.76 at 12 h. The results of animal imaging in vivo showed that CBD NPs could be effectively accumulated in colon tissue. Not only that, CBD showed significant glutathione stimulated release in the presence of 10 mM glutathione at pH 7.4. The results of in vivo animal experiments showed that CBD NPs significantly ameliorated DSS-induced colonic inflammation by modulating the TLR4-NF-κB signaling pathway. Moreover, CBD NPs significantly improved the histological damage of colon in UC mice, increased the expression level of tight junction protein ZO-1, and effectively restored the intestinal mucosal barrier function and intestinal mucosal permeability. More importantly, CBD NPs significantly improved the species composition, abundance and amount of short chain fatty acids of intestinal flora in UC mice, thus effectively maintaining the balance of intestinal flora. The dual-targeted and glutathione-responsive nanoparticles prepared in this study provide a promising idea for achieving targeted delivery of CBD for effective treatment of UC.

Effects of Mactra chinenesis Peptides on Alcohol-Induced Acute Liver Injury and Intestinal Flora in Mice.

Food-borne bioactive peptides have shown promise in preventing and mitigating alcohol-induced liver injury. This study was the first to assess the novel properties of Mactra chinenesis peptides (MCPs) in mitigating acute alcoholic liver injury in mice, and further elucidated the underlying mechanisms associated with this effect. The results showed that MCPs can improve lipid metabolism by modulating the AMPK signaling pathway, decreasing fatty acid synthase activity, and increasing carnitine palmitoyltransferase 1a activity. Meanwhile, MCPs ameliorate inflammation by inhibiting the NF-κB activation, leading to reduced levels of pro-inflammatory cytokines (tumor necrosis factor-α and interleukin-1ß). Additionally, a 16S rDNA sequencing analysis revealed that MCPs can restore the balance of gut microbiota and increase the relative abundance of beneficial bacteria. These findings suggest that supplementation of MCPs could attenuate alcohol intake-induced acute liver injury, and, thus, may be utilized as a functional dietary supplement for the successful treatment and prevention of acute liver injury.

Bovine serum albumin-liposome stabilized high oil-phase emulsion: Effect of liposome ratio on interface properties and stability.

This study aimed to solve the issue of poor lipophilicity of natural bovine serum albumin (BSA) by combining with liposomes (Lips) to stabilize high oil-phase emulsions (HOPEs). The interaction between BSA and Lips was mainly driven by hydrophobic forces, followed by hydrogen bonding. The secondary structure and tertiary structure of BSA were characterized and indicated that the addition of Lips promoted the structural expansion of BSA exposing the hydrophobic groups inside. Interfacial adsorption behaviours were assessed through dynamic interfacial tension, three-phase contact angle, and quartz crystal microbalance with dissipation. These results indicated that BSA-Lips crosslinking improved wettability, promoting adsorption and rearrangement at the oil-water interface, thereby resulting in a dense interfacial layer. The emulsifying efficacy of BSA-stabilized HOPEs also displayed a distinct Lips dependency. Varying the BSA-to-Lips ratio transformed their consistency from flowing to semi-solid, reinforcing the gel network. Under optimal conditions (BSA: Lips = 1:1), the droplet size of BSA-Lips stabilized HOPEs reached a minimum with a highly uniform distribution. Moreover, a 1:1 BSA to Lips ensured outstanding storage, thermal, and centrifugal stability for the HOPEs. This work provides valuable references for the interaction between protein and Lips, guiding the development of highly stable HOPEs stabilizers.

Flavor formation of tilapia byproduct hydrolysates in Maillard reaction.

The Maillard reaction (MR) of tilapia byproduct protein hydrolysates was investigated for the use of byproduct protein as a food ingredient and to mask its fishy odor and bitter flavor. The flavor differences in tilapia byproduct hydrolysates before and after the MR were analyzed to explore the key flavor precursor peptides and amino acids involved in MR. The results suggested that eight key volatile substances, including 2,5-dimethylpyrazine, 2-pentylfuran, hexanal, octanal, nonanal, (E)-2-decenal, decanal, and 1-octen-3-ol contributed most to the MR products group (ROAV > 1). Ten volatile compounds, including 1-octen-3-ol, hexanal, 2-pentylfuran, 2,5-dimethylpyrazine, methyl decanoate, and 2-octylfuran, were the flavor markers that distinguished the different samples (VIP > 1). The four most consumed peptides were VAPEEHPTL, GPIGPRGPAG, KSADDIKKAF, and VWEGQNIVK. Umami peptides and bitter free amino acids (FAAs) were the key flavor precursor peptide and FAAs, respectively. Overall, the hydrolysates of tilapia byproducts with flavor improved by MR are a promising strategy for the production of flavorings.

Novel hydrocolloids synthesized by polyphenols grafted onto chitosan: A promising coating to inhibit PhIP formation during pan-frying of golden pompano fillets.

Hydrocolloids synthesized by gallic acid (GA) and ferulic acid (FA) grafting onto chitosan (CS) were characterized, and their effects on PhIP formation in pan-fried golden pompano were investigated. Spectrograms including nuclear magnetic resonance, Fourier transform infrared spectroscopy and ultraviolet-visible confirmed that GA and FA were successfully grafted onto CS via covalent bonds, with grafting degree of 97.06 ± 2.56 mg GA/g and 93.56 ± 2.76 mg FA/g, respectively. The CS-g-GA and CS-g-FA exerted better solubility and antioxidant activities than CS. For the 8-min pan-fried golden pompano fillets, CS-g-GA and CS-g-FA (0.5 %, m/v) significantly reduced the PhIP formation by 61.71 % and 81.64 %, respectively. Chemical models revealed that CS-g-GA and CS-g-FA inhibited PhIP formation mainly by decreasing the phenylacetaldehyde contents from Maillard reaction and competing with creatinine to react with phenylacetaldehyde. Therefore, it was suggested that CS-g-phenolic acids emerge as novel coating for aquatic products during processing and inhibit heterocyclic amines generation.

Preparation of liposomes by glycolipids/phospholipids as wall materials: Studies on stability and digestibility.

Glycolipids may be potential materials to improve the instability of liposomes during storage and consumption. Curcumin-loaded liposomes with high stability were successfully prepared by glycolipids and phospholipids extracted from tilapia. The physicochemical properties analysed showed that glycolipids enhanced the surface charge of liposomes and the encapsulation ability of curcumin. The enhanced affinity of liposomes for curcumin was attributed to the stronger interaction between the head group of glycolipids and curcumin through hydrogen bonding. As predicted, glycolipids improved the storage stability of liposomes, and the thermal stability of curcumin increased from 35.95% to 54.13%. Moreover, glycolipids could resist the degradation of liposomes in the gastrointestinal tract, reducing the encapsulation efficiency changes of curcumin from 60.67% to 43.63%. Simultaneously, the liposomes formed by glycolipids could more effectively protect nerve cells from oxidative damage. Therefore, the substitution of phospholipids with glycolipids is an effective strategy to improve the stability and bioactivity of liposomes.

Interaction between liposome and myofibrillar protein in surimi: Effect on gel structure and digestive characteristics.

This study investigated the effects of the interaction between liposomes and myofibrillar protein (MP) on tilapia surimi. The strong interaction between liposomes and MP was primarily mediated through hydrogen bonding and hydrophobic interaction. Liposomes caused the unfolding of MP structure, resulting in the decrease of α-helix content and transformation of spatial structure. Notably, the appropriate ratio of liposomes improved the gel properties of tilapia surimi. The water distribution, microstructure, and texture characteristics further confirmed that liposomes strengthened the structure of surimi gel through non-covalent bonds. However, excessive liposomes (1.0 %) weakened gel characteristics and texture. Moreover, the proper ratio of liposomes enhanced the stability of surimi gels during digestion, reducing protein digestibility from 66.0 % to 54.8 %. Curcumin-loaded liposomes in gel matrix notably delayed digestion and improved bioavailability. This delay in digestion was attributed to the ability of liposomes to decrease the interaction between MP and digestive enzymes. This study provides new insight into the application of liposomes in protein-rich food matrixes.

Effects of different drying temperatures on the profile and sources of flavor in semi-dried golden pompano (Trachinotus ovatus).

Drying is an important process that can impart a different flavor to dried fish. The differences and sources of flavor of semi-dried golden pompano at different drying temperatures (40, 48, and 56 °C) were investigated, and the sensory quality, flavor substances, nonenzymatic reactions, and protein degradation were analyzed. Significant flavor differences were observed among different drying temperatures (P < 0.05), with several sensory properties showing superior results in the 40 °C group. Thirteen volatile compounds that contributed to the overall aroma were screened according to the relative odor activity value. Glu (umami taste) and Ala (sweet taste) were identified as key flavor substances based on the taste activity value. Nonanal, hexanal, heptanal, acetoin, pentadecane, and octanal represented the flavor markers. The flavor sources at higher drying temperatures included the joint action of lipid oxidation and the Maillard reaction, while those at lower temperature were lipid oxidation and protein degradation, which increased the aldehyde and free amino acid levels in the product, thus leading to the best flavor.

Emulsion gel stabilized by tilapia myofibrillar protein: Application in lipid-enhanced surimi preparation.

In this study, surimi products rich in lipids were prepared by using myofibril protein (MP) emulsion gel as carriers. The MP emulsion gel (MP concentration, c = 1.5%, oil fraction, ø = 0.68) was prepared by one-step homogenization. The emulsion gel maintained a high elastic modulus (G') after heating and freezing treatment. Confocal laser scanning microscopy revealed that the structure of the emulsion gel was a hybrid network consisting of polymers of cross-linked MP and aggregated protein-stabilized emulsion (W/O/W multiple structures) droplets. The double emulsification of the emulsion gel and MP stabilized the oil droplets in the surimi product, preventing water and oil from leaching out. The microstructure also showed smaller gaps between MPs with increased porosity, while oil droplets were stably embedded in the surimi gel matrix. Moreover, adding MP emulsion gel significantly reduced the surimi gel strength compared to adding oil directly (p < 0.05).

Identification and Characterization of Novel ACE Inhibitory and Antioxidant Peptides from Sardina pilchardus Hydrolysate.

Sardina pilchardus is a valuable source of bioactive peptides with potential applications in functional foods. In this study, we investigated the angiotensin-converting enzyme (ACE) inhibitory activity of Sardina pilchardus protein hydrolysate (SPH) produced using dispase and alkaline protease. Our results showed that the low molecular mass fractions (<3 kDa) obtained through ultrafiltration exhibited more effective ACE inhibition, as indicated by screening with ACE inhibitory activity. We further identified the low molecular mass fractions (<3 kDa) using an LC-MS/MS rapid screening strategy. A total of 37 peptides with potential ACE inhibitory activity were identified based on high biological activity scores, non-toxicity, good solubility, and novelty. Molecular docking was used to screen for peptides with ACE inhibitory activity, resulting in the identification of 11 peptides with higher -CDOCKER ENERGY and -CDOCKER INTERACTION ENERGY scores than lisinopril. The sequences FIGR, FILR, FQRL, FRAL, KFL, and KLF were obtained by synthesizing and validating these 11 peptides in vitro, all of which had ACE inhibitory activity, as well as zinc-chelating capacity. All six peptides were found to bind to the three active pockets (S1, S2, and S1') of ACE during molecular docking, indicating that their inhibition patterns were competitive. Further analysis of the structural characteristics of these peptides indicated that all six peptides contain phenylalanine, which suggests that they may possess antioxidant activities. After experimental verification, it was found that all six of these peptides have antioxidant activities, and we also found that the SPH and ultrafiltration fractions of SPH had antioxidant activities. These findings suggest that Sardina pilchardus may be a potential source of natural antioxidants and ACE inhibitors for the development of functional foods, and using LC-MS/MS in combination with an online database and molecular docking represents a promising, effective, and accurate approach for the discovery of novel ACE inhibitory peptides.

Glycolipids improve the stability of liposomes: The perspective of bilayer membrane structure.

The storage and thermal stability of liposomes, which are amphiphilic carriers, cause very large challenges. However, glycolipid modification may be a potential method to improve the stability of liposomes. In this study, the mechanism by which tilapia head glycolipids improve the stability of liposomes was studied. The head groups of glycolipids and liposomes have a strong interaction (Ka = 633.650 M-1), mainly due to hydrogen bonds, which promote the formation of microstructure domains between glycolipids and liposomes. In addition, glycolipids caused the bilayer structure of liposomes to rearrange, resulting in an increase in the phase transition temperature, tight arrangement of membrane molecules, and increase in membrane thickness (from 2.4 nm to 3.5 nm). Novelty, the formation of microstructure domains helped prevent the liposomes membrane structure from being disrupted during storage and heat. Therefore, glycolipid modification improved the stability of liposomes. This study can provide new insights into the development of high-stability liposomes.

Improving the flavor of tilapia fish head soup by adding lipid oxidation products and cysteine.

Deodorization and umami enhancement are important challenges in promoting and consuming fish products. The aim of this study was to establish whether exogenous addition of oxidized lipids and cysteine can improve the fishy, umami and create a characteristic flavor in tilapia fish head soup. The results revealed that adding oxidized lipids and cysteine enhanced the sensory attributes of fish head soup and promoted the production of pleasant-tasting amino acids and fewer bitter amino acids in the Maillard reaction. Additionally, the combination increased the levels of well-flavored aldehydes, esters, heterocyclic compounds and less hydrocarbons in the fish head soup. Among the 13 volatile compounds screened, nine were identified as characteristic aromas of fish head soup, including nonanal, (E,E)-2,4-decadienal, 1-octen-3-ol, (E)-2-decenal, acetic acid, hexanal, heptanal, 2-octenal, and decanal. Exogenous lipid oxidation products, fatty acid oxidation, and Maillard reaction can improve the aroma and umami of tilapia fish head soup.

Holothuria leucospilota Polysaccharides Improve Immunity and the Gut Microbiota in Cyclophosphamide-Treated Immunosuppressed Mice.

SCOPE: Immunosuppression is one of the major risk factors for a series of diseases, such as tumor, rheumatoid arthritis, and microbial infection. Various natural products have attracted wide attention due to their immunomodulatory activities. Herein, the study investigates the regulation of Holothuria leucospilota polysaccharides (HLP) in immunosuppressed mice. METHODS AND RESULTS: Eight-week-old female BALB/c mice are injected intraperitoneally with cyclophosphamide (80 mg kg-1 body weight day-1 ) to establish the immunosuppressive model. After 12 days of HLP treatment, the immune organ indexes, serum cytokines, and immunoglobulin levels are significantly increased in immunosuppressed mice (p < 0.05). Real-time fluorescent quantitative polymerase chain reaction and western blotting analysis find that HLP improves the immune factors, T-cell markers, and Toll-like receptors (TLR) pathway-related proteins expression. Simultaneously, HLP significantly increases the short-chain fatty acids concentration and regulates the gut microbiota composition (p < 0.05). Furthermore, the metagenomics analysis shows that HLP increases the levels of functional genes involved in amino acid metabolism, carbohydrate metabolism, and growth activity of the gut microbiota. CONCLUSION: HLP intervention improves the mice's immune function, and the beneficial effects are closely associated with intestinal homeostasis regulation and TLR pathway activation. This study suggests the potential for HLP as prebiotics in novel immunopotentiators development.