•High solid concentrations (28%, w/w) facilitate the enzymatic yeast cells hydrolysis.•Higher protein recovery, DH, reducing sugar contents and 5′-nucleotides contents can be achieved at 28% high solid.•The micromorphology of hydrolysed yeast cells can confirm the chemical analysis results.•The hydrolysis efficiency of alcalase for yeast cells is higher than that of papain.To investigate the effect of high solid concentrations (w/w) on the enzymatic hydrolysis of yeast cells, yeast cells were hydrolyzed by papain and Alcalase at 18% and 28% solid concentrations (w/w), respectively. High solid concentrations resulted in high protein recovery, degree of hydrolysis (DH) and contents of reducing sugar and 5′-nucleotides (5′-GMP, 5′-IMP), which indicated a positive effect on the release of yeast intracellular substances. High-performance liquid chromatography (HPLC) analysis showed negligible difference in the molecular mass distribution of peptides. The micromorphology of the hydrolysed yeast cells was studied by transmission electron microscopy (TEM). Result demonstrated that high solid concentrations accelerated the plasmolysis, deformation and lysis of yeast cells. Similarly, addition of NaCl or commercial yeast extract to the yeast cell suspension (18% w/w) improved the DH. Therefore, high concentrations of NaCl or yeast cell hydrolysates in solutions with high solid concentrations could damage the yeast cell membrane, leading to effective contact between the enzyme and the substrate and consequent increase in the hydrolysis efficiency. Nevertheless, the hydrolysis mechanism remains unknown and must be further investigated.Download high-res image (161KB)Download full-size image

With lower cell toxicity and higher specificity, novel vaccines have been greatly developed and applied to emerging infectious and chronic diseases. However, due to problems associated with low immunogenicity and complicated processing steps, the development of novel vaccines has been limited. With the rapid development of bio-technologies and material sciences, nanomaterials are playing essential roles in novel vaccine design. Incorporation of nanomaterials is expected to improve delivery efficiency, to increase immunogenicity, and to reduce the administration dosage. The purpose of this review is to discuss the employment of nanomaterials, including polymeric nanoparticles, liposomes, virus-like particles, peptide amphiphiles micelles, peptide nanofibers and microneedle arrays, in vaccine design. Compared to traditional methods, vaccines made from nanomaterials display many appealing benefits, including precise stimulation of immune responses, effective targeting to certain tissue or cells, and desirable biocompatibility. Current research suggests that nanomaterials may improve our approach to the design and delivery of novel vaccines.

The walnut peptides and zinc ions were combined to generate a walnut peptides-zinc complex (WP1-Zn) with enhanced antiproliferative ability as well as reduced toxicity. The result indicated that Zn ions were successfully combined with WP1 through Zn–N and Zn–O covalent bonds. WP1-Zn compounds exhibited strong antiproliferative ability against the selected human cell lines, especially MCF-7 cells, whose survival rate reduced to 20.02% after exposure to 300 μg/mL of WP1-Zn for 48 h. WP1-Zn inhibited MCF-7 cell proliferation through inducing cell apoptosis and cell cycle arrest. The results indicated that WP1-Zn induced MCF-7 cell apoptosis via the ROS triggered mitochondrial-mediated pathway and cell surface receptor-mediated pathway. Our work is the first attempt to elucidate the synergic effect of novel walnut peptides and Zn and with the hope of better understanding the antiproliferative action of bioactive peptides and a zinc complex and support the potential application of WP1-Zn as a functional food ingredient or complementary medicine.

A polysaccharide fraction, here called POP1, was purified from the leaves of Platycladus orientalis (L.) Franco by water extraction and alcohol precipitation. Physicochemical characterization indicated that POP1 had a relative molecular weight of 8.10 × 103 Da and consisted of rhamnose (5.74%), arabinose (12.58%), mannose (10.97%), glucose (64.96%), and galactose (6.55%). The main linkage types of POP1 consisted of (1→5)-linked α-l-Ara, (1→3)-linked α-l-Man, (1→6)-linked β-l-Rha, (1→4)-linked α-d-Glc, (1→6)-linked α-d-Glc, (1→6)-linked β-d-Gal, (1→3,6)-linked β-d-Gal, and termination with α-l-Man and α-d-Glc residues based on periodate oxidation, Smith degradation, methylation, and NMR analysis. POP1 exhibited excellent immunostimulating activity by enhancing macrophage NO, TNF-α, IL-6, and IL-12 secretion and activating related mRNA expression. Besides, POP1 showed significant anti-HBV activity through inhibiting the expression of HBsAg (IC50 = 1.33 ± 0.12 mg/mL) and HBeAg (IC50 = 1.67 ± 0.13 mg/mL) and interfering with the HBV DNA replication (IC50 = 0.80 ± 0.03 mg/mL). The present study suggested that POP1 could be used as immunoregulatory agent in functional foods for the prevention of HBV infection.

Dictyophora indusiata polysaccharide (DP1) was successfully chelated with zinc chloride to achieve its enhanced antiproliferative activity. The obtained DP1–Zn complex showed significant antiproliferative activity (18.1 ± 2.84% viability of MCF-7 cells at 250 μg/mL) on a group of human cancer cell lines through induction of apoptosis. The pro-apoptotic action of DP1–Zn was confirmed by morphological changes including chromatin condensation, DNA breakage, and S phase cell cycle arrest in human breast adenocarcinoma cells (MCF-7). The DP1–Zn-induced apoptotic pathways were characterized by the activation of caspases-3, -8, and -9, mitochondrial dysfunction, and reactive oxygen species (ROS) overproduction (305 ± 7.06% production of control at 250 μg/mL). This study suggested that DP1–Zn can be developed as a candidate for cancer treatment and prevention, especially human breast adenocarcinoma.

A novel polysaccharide, here named DP1, was isolated from the fruiting body of Dictyophora indusiata using a water extraction method. Structure characterization revealed that DP1 had an average molecular weight of 1132 kDa and consisted of glucose (56.2%), galactose (14.1%), and mannose (29.7%). The main linkage type of DP1 were proven to be (1 → 3)-linked α-l-Man, (1 → 2,6)-linked α-d-Glc, (1 → 6)-linked β-d-Glc, (1 → 6)-linked β-d-Gal, and (1 → 6)-linked β-d-Man by periodate oxidation–Smith degradation and nuclear magnetic resonance analysis. The immunostimulating assay indicated that DP1 could significantly promote macrophage NO, TNF-α, and IL-6 secretion in murine RAW 264.7 cells involving complement receptor 3 (CR3). The immune activities of DP1 were quite stable under thermal processing (100, 121, and 145 °C). Besides, DP1 retained stability after acidic/alkline treatment (pH 4.0–10.0), which enabled it to be an ideal complementary medicine or functional food for therapeutics of hypoimmunity and immunodeficiency diseases.

Phytohemagglutin (PHA), purified from red kidney beans (Phaseolus vulgaris) by Affi-Gel blue affinity chromatography, was subjected to ultrahigh-pressure (UHP) treatment (150, 250, 350, and 450 MPa). The purified PHA lost its hemagglutination activity after 450 MPa treatment and showed less pressure tolerance than crude PHA. However, the saccharide specificity and α-glucosidase inhibition activity of the purified PHA did not change much after UHP treatment. Electrophoresis staining by periodic acid–Schiff (PAS) manifested that the glycone structure of purified PHA remained stable even after 450 MPa pressure treatment. However, electrophoresis staining by Coomassie Blue as well as circular dichroism (CD) and differential scanning calorimetry (DSC) assay proved that the protein unit structure of purified PHA unfolded when treated at 0–250 MPa but reaggregates at 250–450 MPa. Therefore, the hemagglutination activity tends to be affected by the protein unit structure, while the stability of the glycone structure contributed to the remaining α-glucosidase inhibition activity.

The intracellular antioxidant activities of diosmetin were evaluated by cellular antioxidant activity (CAA) assay, 2,2-azobis(2-amidinopropane) dihydrochloride (AAPH)-induced erythrocyte hemolysis assay and cupric chloride (CuCl2)-induced plasma oxidation assay. The results showed that diosmetin exhibits strong cellular antioxidant activity (EC50 = 7.98 μmol, CAA value = 58 μmol QE/100 μmol). It was also found that diosmetin treatment could effectively attenuate AAPH-induced erythrocyte hemolysis (91.0% inhibition at 100 μg/mL) and CuCl2-induced plasma oxidation through inhibition of intracellular reactive oxygen species (ROS) generation. Diosmetin could significantly restore AAPH-induced increase of intracelluar antioxidant enzyme (SOD, GPx, and CAT) activities to normal levels, as well as inhibit intracellular malondialdehyde (MDA) formation. Thus, the intracellular antioxidant detoxifying mechanism of diosmetin is associated with both nonenzymatic and enzymatic defense systems.

Red kidney beans were subjected to high hydrostatic pressure (HHP) treatment (50, 150, 250, 350, 450 MPa) and phytohaemagglutinin (PHA) was then extracted by affinity chromatography. It appeared that HHP treatment could increase crude extract yield and decrease its haemagglutination activity. For purified samples, PHA yield was not affected at pressures <450 MPa while the haemagglutination activity was noticeably reduced at 450 MPa. The structural changes were investigated using electrophoresis, size exclusion chromatography (SEC), Fourier transform infrared (FTIR) and differencial scanning calorimetry (DSC). Electrophoresis and SEC profiles revealed a new high molecular weight polymer after 450 MPa treatment. At pressures <450 MPa, FTIR showed an increase in β-sheet structure and a decrease in α-helix. At 450 MPa, the bands at 1688 cm−1, representing aggregate strands and random coils, increased. The conclusions are that pressures <450 MPa can cause PHA unfolding and induce PHA aggregation at 450 MPa.Highlights► HHP treatment could decrease the haemagglutination activity of PHA. ► HHP could interfere with the secondary structures of PHA. ► HHP changed the thermal behaviour of PHA. ► FTIR and DSC proved PHA unfolding caused by low pressure <450 MPa. ► FTIR and DSC proved PHA aggregation induced by high pressure >450 MPa.

Peanut protein isolate (PPI) was treated by high-pressure microfluidization (40, 80, 120, and 160 MPa) and/or transglutaminase (TGase) cross-linking. It was found that individual microfluidization at 120 MPa was more effective in improving the solubility, emulsifying properties, and surface hydrophobicity of PPI than at other pressures (e.g., 40, 80, or 160 MPa). Individual TGase cross-linking also effectively changed the physicochemical and functional properties of PPI. Microfluidization (120 MPa) or TGase cross-linking caused the unfolding of PPI structure, resulting in the decrease of α-helix and β-turns levels and the increase of β-sheet and random coil levels, as proved by Fourier transform infrared (FTIR) and circular dichroism (CD) spectra. Compared with individual treatments, microfluidization followed by TGase cross-linking significantly (p < 0.05) improved the emulsion stability during long-term storage (20 days). Moreover, the combined treatments led to looser structure of PPI and resulted in more obvious changes in physicochemical properties.

With lower cell toxicity and higher specificity, novel vaccines have been greatly developed and applied to emerging infectious and chronic diseases. However, due to problems associated with low immunogenicity and complicated processing steps, the development of novel vaccines has been limited. With the rapid development of bio-technologies and material sciences, nanomaterials are playing essential roles in novel vaccine design. Incorporation of nanomaterials is expected to improve delivery efficiency, to increase immunogenicity, and to reduce the administration dosage. The purpose of this review is to discuss the employment of nanomaterials, including polymeric nanoparticles, liposomes, virus-like particles, peptide amphiphiles micelles, peptide nanofibers and microneedle arrays, in vaccine design. Compared to traditional methods, vaccines made from nanomaterials display many appealing benefits, including precise stimulation of immune responses, effective targeting to certain tissue or cells, and desirable biocompatibility. Current research suggests that nanomaterials may improve our approach to the design and delivery of novel vaccines.