Linear Glucan Research Articles

A short linear glucan nanocomposite hydrogel formed by in situ self-assembly with highly elastic, fatigue-resistant and self-recovery

Polyacrylamide (PAM) hydrogels are widely used in wide-ranging applications in biology, medicine, pharmaceuticals and environmental sectors. However, achieving the requisite mechanical properties, fatigue resistance, self-recovery, biocompatibility, and biodegradability remains a challenge. Herein, we present a facile method to construct a nanocomposite hydrogel by integrating short linear glucan (SLG), obtained by debranching waxy corn starch, into a PAM network through self-assembly. The resulting composite hydrogel with 10 % SLG content exhibited satisfactory stretchability (withstanding over 1200 % strain), along with maximum compressive and shear strengths of about 490 kPa and 39 kPa at 90 % deformation, respectively. The hydrogel demonstrated remarkable resilience and could endure repeated compression and stretching. Notably, the nanocomposite hydrogel with 10 % SLG content exhibited full stress recovery at 90 % compression deformation after 20 s, without requiring specific environmental conditions, achieving an energy dissipation recovery rate of 98 %. Meanwhile, these hydrogels exhibited strong adhesion to various soft and hard substrates, including skin, glasses and metals. Furthermore, they maintain solid integrity at both 37 °C and 50 °C after swelling equilibrium, unlike traditional PAM hydrogels, which exhibited softening under similar conditions. We hope that this PAM-SLG hydrogel will open up new avenues for the development of multifunctional electronic devices, offering enhanced performance and versatility.

Enzymatic Synthesis of α-Glucan Microparticles Using Amylosucrases from Bifidobacterium Species and Its Physicochemical Properties.

α-Glucan microparticles (GMPs) have significant potential as high-value biomaterials in various industries. This study proposes a bottom-up approach for producing GMPs using four amylosucrases from Bifidobacterium sp. (BASs). The physicochemical characteristics of these GMPs were analyzed, and the results showed that the properties of the GMPs varied depending on the type of enzymes used in their synthesis. As common properties, all GMPs exhibited typical B-type crystal patterns and poor colloidal dispersion stability. Interestingly, differences in the physicochemical properties of GMPs were generated depending on the synthesis rate of linear α-glucan by the enzymes and the degree of polymerization (DP) distribution. Consequently, we found differences in the properties of GMPs depending on the DP distribution of linear glucans prepared with four BASs. Furthermore, we suggest that precise control of the type and characteristics of the enzymes provides the possibility of producing GMPs with tailored physicochemical properties for various industrial applications.

Mechanistic enhancement of emulsification function in zein/pectin complex nanoparticles by short linear glucan

The objective of this research was to create stable Pickering emulsions (PEs) by manipulating the structure of the interface through the control of interfacial properties, self-assembly, and packing characteristics of zein particles, utilizing the interaction between short linear glucan (SLG) and pectin. The scientists successfully synthesized ternary complex particles designated as SZPPs (abbreviated for SLG/Zein/Pectin), displaying three-phase contact angles ranging from 83.14° to 87.32°. SZPPs were effectively anchored at the oil-water interface, resulting in a robust and well-organized arrangement, as confirmed by the combination of Langmuir−Blodgett and Scanning Electron Microscope (LB-SEM) and confocal laser scanning microscopy techniques. This arrangement led to the formation of a permeating three-dimensional network of oil droplets, which greatly contributed to the creation of stable PEs. These PEs exhibited unique properties such as viscoelasticity, remarkable thixotropy, and excellent long-term storage stability. One noteworthy advantage of these engineered PEs was their ability to shield curcumin from degradation caused by UV radiation, thereby enhancing the oxidative stability of the emulsions. This protective effect ensures that the curcumin remains intact and active over time. Considering these advantageous properties, these engineered PEs hold great promise for various applications in the food and pharmaceutical industries.

Optimization and analysis of dual-enzymatic synthesis for the production of linear glucan

Linear α-glucan (LG), a linear polymer linked by α-1,4 bonds, has received increasing attention for its potential applications in synthetic polymer production. Notably, the functionality of LG is strongly influenced by its degree of polymerization (DP). In this study, SP and GP were successfully constructed and expressed. The reaction of enzymatic co-polymerization into LG was investigated. The preferred reaction was carried out at 37 °C and pH 7.4 for 72 h, with a maximum conversion rate of 25 %. Afterwards, two approaches were used to modulate the molecular structures of LGs. Firstly, LGs with distinct molecular weights ranging from 1062.33 ± 16.04 g/mol to 5679 ± 80.29 g/mol were obtained by adjusting the substrate/primer ratio during the LG synthesis process. Secondly, two distinct products could be produced by altering the enzyme addition method: short-chain LG with a DP < 10 (64.34 ± 0.54 %) or long-chain LG with a DP > 45 (45.57 ± 2.18 %). Additionally, theoretical synthesis model was constructed which subdivided the reaction into three stages to evaluate this dual-enzyme cooperative system. These findings have significant implications in promoting the application of LG in the fields of biomedicine and material science.

Structural Characteristics, Physicochemical Properties, and Digestibility Analysis of Resistant Starch Type-V Prepared from Debranched Corn Starch and Fatty Acid Complexation.

Corn starch was gelatinized and treated with a metagenomic type 1 pullulanase (PulM), increasing the proportion of linear glucan chains. The debranched corn starch (DCS), containing amylose helices, was subjected to complexation with fatty acid molecules at moderate temperatures (50-60 °C). The amylose-lipid complexes prepared using saturated fatty acids, e.g., capric acid (CA) and lauric acid (LA), displayed higher CI values as compared to that of unsaturated fatty acid compounds, e.g., undecylenic acids (UAs) and oleic acid (OA). The DCS-fatty acid complex was estimated to contain about 14% of rapidly digested starch (RDS), 26% of slowly digested starch (SDS), and 60% of resistant starch V (RS-5). RS-5 samples exhibited high resistance toward digestive enzymatic hydrolysis. The surface microdetails of RS-5 were examined by scanning electron microscopy (SEM), depicting small spherulite-like structural aggregates. X-ray diffraction pattern analysis estimated about 46% of the crystallinity of RS-5. Thermal attributes of RS-5 were examined by thermogravimetric analysis (TGA) and differential scanning calorimetry (DSC) analysis, depicting the increase in melting enthalpies after the complexation of fatty acid molecules with debranched corn starch. Comparative DSC thermograms divulged a relatively higher stability of RS-5 as compared to that of RS-3. The findings advocated the potentiality of RS-5 (nondigestible DCS-LA complex) as a functional, valuable ingredient in the food industry.

Structural characterization of different starch-fatty acid complexes and their effects on human intestinal microflora.

Resistant starch type 5 (RS5), a starch-lipid complex, exhibited potential health benefits in blood glucose and insulin control due to the low digestibility. The effects of the crystalline structure of starch and chain length of fatty acid on the structure, in vitro digestibility, and fermentation ability in RS5 were investigated by compounding (maize, rice, wheat, potato, cassava, lotus, and ginkgo) of different debranched starches with 12-18C fatty acid (lauric, myristic, palmitic, and stearic acids), respectively. The complex showed a V-type structure, formed by lotus and ginkgo debranched starches, and fatty acid exhibited a higher short-range order and crystallinity, and lower in vitro digestibility than others due to the neat interior structure of more linear glucan chains. Furthermore, a fatty acid with 12C (lauric acid)-debranched starches complexes had the highest complex index among all complexes, which might be attributed to the activation energy required for complex formation increased with the lengthening of the lipid carbon chain. Therefore, the lotus starch-lauric acid complex (LS12) exhibited remarkable ability in intestinal flora fermentation to produce short-chain fatty acid (SCFAs), reducing intestinal pH, and creating a favorable environment for beneficial bacteria.

Effects of polymerization degree of short linear glucan on the properties of Pickering emulsion stabilized by short linear glucan /zein nanoparticles

Here we fabricated short linear glucan/zein (SLG/zein) complex nanoparticles using the reverse dropwise method and studied how the average SLG polymerization degree ( DPn̅) effects the complex nanoparticles’ physicochemical properties and emulsion stability. Using fourier transform infrared and fluorescence spectroscopies, we found that electrostatic interactions and hydrogen bonding were the main driving forces for complex nanoparticle formation. Furthermore, our wettability results showed that SLG with a low DPn̅ can neutralize zein’s hydrophobic properties, making the complex nanoparticles more suitable for stabilizing a Pickering emulsion. We then loaded the nanoparticles with curcumin and achieved a maximum encapsulation efficiency of 70.45%. After subjecting the emulsion to a three-step in vitro digestion process, we found that the complex nanoparticles with lower DPn̅values provided better protection against the biochemical conditions of gastric fluid, resulting in greater curcumin bioaccessibility. These results suggest that the SLG/zein complex nanoparticles have great potential to encapsulate bioactive ingredients as a Pickering emulsifier.

Physicochemical and In Vitro Starch Residual Digestion Structures of Extruded Maize and Sorghum Starches Added with Sodium Stearoyl Lactylate.

This research aimed to characterize the physicochemical, in vitro digestion, and structural features of digestion residues of maize and sorghum starches subjected to thermoplastic extrusion, along with the influence of Sodium Stearoyl Lactylate (SSL), to obtain improved starches for food applications and to understand their behavior when consumed as a food ingredient. The morphology of the extruded materials showed remanent starch granules when SSL was used. A higher amount of medium and large linear glucan chains were found in these particles, influencing higher thermal stability (ΔH ≈ 4 J/g) and a residual crystallinity arrangement varying from 7 to 17% in the extrudates. Such structural features were correlated with their digestibility, where slowly digestible starch (SDS) and resistant starch (RS) fractions ranged widely (from 18.28 to 27.88% and from 0.13 to 21.41%, respectively). By analyzing the data with a Principal component analysis (PCA), we found strong influences of B2 and B3 type chains on the thermal stability of the extrudates. The amylose and smaller glucan chains (A and B1) also significantly affected the emulsifying and foam stability properties. This research contributes to the molecular knowledge of starch in extruded products with broad food applications.

Structure and effect onBifidobacterium longumof exopolysaccharide produced byWeissella cibariausing cost-effective substrates

AbstractMaize straw, an abundant agricultural residue, has potential as an eco-friendly resource. The aim of this study was to investigate the structure and effect on Bifidobacterium longum of exopolysaccharide produced by Weissella cibaria using maize straw as a carbon source. In this work, an exopolysaccharide-producing strain was isolated from vegetable juice, which was identified as W. cibaria. Our results demonstrated that exopolysaccharide produced by W. cibaria is a linear glucan containing an α-1,6 glycosidic bond with a molecular weight of 9462 Da. In addition, scanning electron microscopy of exopolysaccharide from W. cibaria revealed the flake structures. In vitro assays showed that the exopolysaccharide from W. cibaria has the ability to stimulate the proliferation of B. longum strain and its metabolism to produce acetic acid. This work provides a scientific reference for the potential application of maize straw and a new insight into an eco-friendly carbon source to produce exopolysaccharide.

An exopolysaccharide from Lactobacillus pentosus YY-112: structure and effect on the human intestinal microbiota.

The development of novel prebiotics, which could regulate the intestinal microbiota, may help prevent and treat intestinal diseases. Here, we studied a homogeneous polysaccharide, LPE-2, produced by Lactobacillus pentosus YY-112 during fermentation. Methylation and gas chromatography-mass spectrometry analysis, combined with nuclear magnetic resonance results, suggested that the structural unit of LPE-2 comprises a branched mannan moiety and a linear glucan moiety. In vitro simulated intestinal fermentation showed that LPE-2 reduced harmful intestinal gas production and promoted short-chain fatty acid production (especially propionic acid). Moreover, it reduced the relative abundance of Escherichia-Shigella, increased that of Bifidobacterium and Lactobacillus, and had a stronger regulatory effect on intestinal flora in women than in men. The potential sex-specific prebiotic effects of LPE-2 on human intestinal health, were possibly related to its mannan branch with (1→2) and (1→3) linkages and backbones with flexible α configurations, which are sheared and degraded/utilized easier by Bifidobacterium and Lactobacillus.

Biobased ionic liquids as solvents of paramylon

Paramylon is a linear glucan produced by Euglena gracilis microalgae as water insoluble intracellular granules. Its macromolecular structure contains only β(1–3) linkages, resulting in a stable triple helix conformation and a high crystallinity. It is soluble in very few solvents such as DMSO, harsh alkaline or acid solutions and imidazolium ionic liquids. In this paper we prepared biosourced more ecofriendly ionic liquids which can tolerate high amounts of water as a co-solvent and which are recyclable. The solubility of the paramylon was then evaluated depending of the nature of the ionic liquid and the structure and thermal properties of the biopolymer before and after solubilisation and regeneration were also discussed.

Fabrication and characterization of self-assembled whey protein isolate/short linear glucan core–shell nanoparticles for sustained release of curcumin

The aim of this research was to prepare a bistratal nanocomplex with a high loading capacity (LC) and harsh environment stability for controlled release of curcumin (Cur) in gastrointestinal conditions. Whey protein isolate (WPI)/short linear glucan (SLG) core–shell nanoparticles were fabricated by self-assembly for the delivery of Cur. The results showed that Cur@WPI@SLG nanoparticles had a relatively high LC (12.89 %) and small particle size (89.4 nm). The nanocomplex remained relatively stable in extreme pH conditions (2–4 and 8–10), high temperatures (60–70 °C), and ionic strength (<400 mM). Core–shell nanostructures facilitated the sustained release of Cur in simulated gastrointestinal conditions. In addition, the nanocomplex had little cytotoxicity at high concentrations, yet significantly enhanced the DPPH scavenging activity and reducing power of Cur. This delivery system will significantly improve the sustained release effect of Cur and broaden the application of hydrophobic nutrients in foods.

Effect of Debranching and Differential Ethanol Precipitation on the Formation and Fermentation Properties of Maize Starch-Lipid Complexes.

The objective of this study was to investigate the effect of starch debranching followed by differential ethanol precipitation on the formation and in vitro fermentation of starch-lipid complexes. Three groups of linear glucan chains, with a degree of polymerization (DP) of 383∼2950, 37∼75, and 3∼8, were obtained after debranched maize starch (DMS) was fractionated by differential ethanol precipitation. The glucan fraction with DP 383∼2950 formed only type IIb complexes with lauric acid (LA), whereas the fraction with DP 37∼75 formed predominantly type Ia complexes. The glucan faction with DP 8∼32 did not form V-complexes with LA. In vitro fermentation of the type IIb complexes with human fecal samples promoted the proliferation of butyrate-producing bacteria Megamonas, Blautia, and Megasphaera and resulted in a larger amount of butyrate and total short-chain fatty acids being produced than in similar fermentations of the maize starch-LA complex, DMS-LA complex, and fructo-oligosaccharides. This study showed that starch-lipid complexes with a more stable type IIb crystallite resulted in a greater production of butyrate.

Encapsulation of Curcumin in a Ternary Nanocomplex Prepared with Carboxymethyl Short Linear Glucan-Sodium-Caseinate-Pectin Via Electrostatic Interactions.

This work chemically modified short linear glucan (SLG) by introducing a surface carboxymethyl group to obtain carboxymethylated SLG (CMSLG), then prepared CMSLG-based ternary nanocomplex particles based on electrostatic interactions with sodium-caseinate (NaCas) and pectin. These nanocomplex particles are homogeneous, generally exhibiting sizes of<200nm with spherical shape and negative surface charge. In addition, the results showed the increase in both the mass ratio of CMSLG and NaCas and the synthesis temperature can improve the colloidal stability of nanocomplex particles when they are exposed to simulated gastrointestinal fluids containing digestive enzymes. Moreover, nanocomplex particles have an exceptional capability to encapsulate curcumin, and this encapsulation efficiency increased as the mass ratios of CMSLG and NaCas were increased. The study also investigated the antioxidant activity and in vitro release properties of curcumin encapsulated by nanocomplex particles and found that CMSLG/NaCas/pectin had improved higher ABTS radical scavenging capacity and allowed for the controlled, sustained release of curcumin in simulated gastrointestinal fluid within 6hours. Thus, this study provides new insights into the design of a CMSLG-based ternary nanocomplex and its use as a potential oral delivery system for lipophilic bioactive compounds. PRACTICAL APPLICATION: Curcumin, as a sort of natural polyphenolic compound, has many physiologic functions such as anti-oxidation, anticancer, and prevention of Alzheimer's disease. However, the application of the curcumin has been limited by its poor water solubility and unstable physicochemical property. To solve this problem, the nanotechnology has been used to prepare the nano-delivery carriers for curcumin. This work prepared a ternary nanoparticle based on the carboxymethyl short linear glucan, sodium-caseinate, and pectin. The ternary nanoparticle can achieve a higher encapsulation efficiency for curcumin. In addition, the ternary nanoparticle can enhance the ABTS radical scavenging capacity and provided control and sustained release of curcumin in the simulated gastrointestinal fluid.

The structures of two polysaccharides from Angelica sinensis and their effects on hepatic insulin resistance through blocking RAGE

This study found two novel homogeneous polysaccharides from Angelica sinensis, APS-1I and APS-2II, binding to RAGE with a dissociation constant of 2.02 ± 0.2 and 85.92 ± 0.2 μM, respectively. APS-1I is a 17.0 kDa heteropolysaccharide, whose backbone is composed of α-1,6-Glcp, α-1,3,6-Glcp, α-1,2-Glcp, α-1,4-Galp, and α-1,3-Rhap, and whose two branches contain α-1,3,5-Araf, α-1,3-Araf, α-1,4-Galp, β-1,3-Galp, and β-1,4-Glcp. APS-2II is a 10.0 kDa linear glucan, that contains α-1,6-Glcp, α-1,3-Glcp, α-1,2-Glcp, and α-T-Glcp. In vitro, APS-1I demonstrated better promotion on glucose absorption and stronger repression on p-IRS-1 (Ser307), p-IRS-2 (Ser731), p-JNK, and p-P38 than APS-2II in insulin resistance (IR)-HepG2 cells. Furthermore, APS-1I treatment couldn't further decrease the inhibition on the phosphorylation of JNK and P38 produced by RAGE siRNA in IR-HepG2 cells. In vivo, APS-1I markedly improved IR and reversed the livers RAGE-JNK/p38-IRS signaling in high-fat-diet and streptozotocin-induced diabetic rats, suggesting that APS-1I could be a potential agent for improving IR in type 2 diabetes.

Fabrication, Structure and Functional Characterizations of pH-Responsive Hydrogels Derived from Phytoglycogen.

The pH-responsive hydrogels were obtained through successive carboxymethylation and phosphorylase elongatation of phytoglycogen and their structure and functional characterizations were investigated. Phytoglycogen (PG) was first carboxymethylated to obtain carboxymethyl phytoglycogen (CM-PG) with degree of substitution (DS) at 0.15, 0.25, 0.30, and 0.40, respectively. Iodine staining and X-ray diffraction analysis suggested that the linear glucan chains were successfully phosphorylase-elongated from the non-reducing ends at the CM-PG surface and assembled into the double helical segments, leading to formation of the hydrogel. The DS of CM-PG significantly influenced elongation of glucan chains. Specifically, fewer glucan chains were elongated for CM-PG with higher DS and the final glucan chains were shorter, resulting in lower gelation rate of chain-elongated CM-PG and lower firmness of the corresponding hydrogels. Scanning electron microscope observed that the hydrogels exhibited a porous and interconnected morphology. The swelling ratio and volume of hydrogels was low at pH 3–5 and then became larger at pH 6–8 due to electrostatic repulsion resulting from deprotonated carboxymethyl groups. Particularly, the hydrogel prepared from chain-elongated CM-PG (DS = 0.25) showed the highest sensitivity to pH. These results suggested that phosphorylase-treated CM-PG formed the pH-responsive hydrogel and that the elongation degree and the properties of hydrogels depended on the carboxymethylation degree. Thus, it was inferred that these hydrogels was a potential carrier system of bioactive substances for their targeted releasing in small intestine.

Characterization of Cationic Modified Short Linear Glucan and Fabrication of Complex Nanoparticles with Low and High Methoxy Pectin.

In this study, we chemically modified the short linear glucan (SLG) using the 3-chloro-2-hydroxypropyl trimethylammonium chloride to introduce a positive surface charge via cationization (CSLG). We then prepared CSLG-based binary nanocomplex particles through electrostatic interactions with low and high methoxyl pectin. The two new types of binary nanocomplex were comprehensively characterized. It was found that the nanocomplex particles showed a spherical shape with the particle size of <700 nm, smooth surface, homogeneous distribution, and negative surface charge. Fourier transform infrared spectroscopy (FTIR) revealed that the driving forces to form nanocomplex were primarily electrostatic interactions and hydrogen bonding. In addition, increasing the CSLG concentration in the nanocomplex significantly enhanced both thermal stability and digestive stability. By comparing the two complex nanoparticles, the HMP-CSLG has a larger particle size and better stability under the GI condition due to the high content of the methoxy group. Additionally, the HMP-CSLG nanoparticle has a higher encapsulation efficiency and slower release rate under simulated gastrointestinal fluid for tangeretin compared with the LMP-CSLG. These results provide new insights into designing the CSLG-based nanocomplex as a potential oral delivery system for nutraceuticals or active ingredients.

Structures and anti-melanoma activities of two polysaccharides from Angelica sinensis (Oliv.) Diels

Polysaccharide is one of the necessary macromolecules in life activities, and it is also a very promising natural product for tumor prevention and treatment. In this study, two homogeneous polysaccharides (APS-4I and APS-4II) were isolated from Angelica sinensis (Oliv.) Diels. APS-4I was a linear glucan with molecular weight of 16.1 kDa, which was composed of 88.4% α-1,6-Glcp, 4.1% α-1,2-Glcp, 3.9% α-1,3-Glcp, and 2.8% α-T-Glcp. APS-4II was a novel polysaccharide with molecular weight of 11.1 kDa, which consisted of 55.4% α-1,6-Glcp, 10.4% α-1,3,5-Araf, 8.7% α-T-Araf, 9.2% α-1,5-Araf, 4.0% α-1,3-Araf, 3.6% α-1,4-Galp, and 9.1% β-1,3-Galp. NMR results demonstrated that APS-4II has a backbone composed of →6)-α-Glcp-(1 → 6)-α-Glcp-(1 → 5)-α-Araf-.(1 → 5)-α-Araf-(1 → 3,5)-α-Araf-(1 → 3)-β-Galp-(1 → 3)-β-Galp-(1 → 4)-α-Galp-(1 → 3)-α-Araf-(1 → 3,5)-α-Araf-(1→. Both APS-4I and APS-4II inhibited the tumor growth of B16-bearing mice, and the suppressive effect of APS-4II reached 64.7 ± 7.3%. Meanwhile, there were higher lymphocyte numbers and the levels of IL-2, IFN-γ, and TNF-α in peripheral blood of APS-4II-treated mice than those in APS-4I-treated mice. Furthermore, APS-4II showed a higher inhibitory effect on the proliferation of B16 cells and stronger promoting effects on the proliferation of splenocytes, the phagocytosis of peritoneal macrophages, and the cytotoxicity of NK cells. These results demonstrated that APS-4II could be a promising therapeutic agent for melanoma.

Production, Characterization, and Antioxidant Activities of an Exopolysaccharide Extracted from Spent Media Wastewater after Leuconostoc mesenteroides WiKim32 Fermentation.

Bacterial exopolysaccharides (EPSs) are important alternatives to plant polysaccharides in fermented products and exhibit antioxidant activity, which is particularly desirable for functional foods. This study evaluated the use of spent media wastewater (SMW) derived from kimchi fermentation for the production of an EPS and analyzed the characterization and antioxidant activity of the resulting EPS. The EPS concentration and conversion yields of sequential purification were 7.7–9.0 g/L and 38.6–45.1%, respectively. Fourier transform infrared spectra and NMR spectra indicated that the EPS was a linear glucan with α-(1 → 6) linkages. The EPS also exhibited thermal tolerance to high temperatures. In vitro antioxidant activity analyses indicated the scavenging activity on 1,1-diphenyl-2-picrylhydrazyl (DPPH) radicals, thiobarbituric acid reactance (TBAR), and ferric ion reducing antioxidant power (FRAP) values of 71.6–79.1, 28.2–33.0%, and 0.04–0.05 mM FeCl3, respectively. These results reveal that the EPS extracted from SMW has potential as a thermally tolerant, nontoxic, and natural antioxidant for industrial applications.

Characterization, prebiotic and immune-enhancing activities of rhamnogalacturonan-I-rich polysaccharide fraction from molokhia leaves

Plant-derived polysaccharides possess potential health benefits that improve intestinal health and the immune system. Molokhia leaves have a large amount of mucilage polysaccharide; in the present study, crude polysaccharide extract was prepared from molokhia leaves. The molecular weight of molokhia leaf polysaccharide fraction (MPF) was estimated to be 51.2 × 103 Da. Polysaccharide was methylated and the structure of MPF was mainly composed of rhamnogalacturonan-I structure with side chains, such as galactans and linear glucan (starch), as shown by GC–MS analysis. To study the biofunctional effects of MPF, its prebiotic and intestinal immune-enhancing activities were assayed in vitro. MPF exhibited good prebiotic activity, as shown by its high prebiotic scores, and increased contents of total short-chain fatty acids on five probiotic strains. In addition, MPF showed immune-enhancing activity on Peyer's patches, as revealed by the high bone marrow cell proliferating activity and production of immunoglobulin A and cytokines. These results demonstrate that MPF may be a potential beneficial prebiotic and intestinal immune-enhancer, which may have wide implications in the food industry.