Functional Food Components Research Articles

Ex vivo and in vitro assessment of anti-inflammatory activity of seed β-conglutin proteins from Lupinus angustifolius

The development of functional food ingredients using legume seed proteins has potential for nutraceutical use. We purified recombinant β-conglutin proteins (rβ1 to rβ4, and rβ6) from narrow-leafed lupin using affinity–chromatography, and evaluated their anti-inflammatory activity using ex vivo and in vitro systems. rβ1, rβ3, and rβ6 produced lower levels of pro-inflammatory mediators such as nitric oxide (about –34.0–fold in all cases), iNOS mRNA (−7.15, −7.97, −7.41–fold), interleukin 1β (−12.05, −11.64, −12.16–fold), chemokine CCL5 (−16.0, −18.0, −19.0–fold), and cytokines including TNF-α, INF-γ, IL-1β, IL-2, IL-6, IL-8, and IL-12 (β1: −28, −100, −8, −2, −49, −45, −127–fold; β3: −22, −400, −9, −3, −33, −10, −2.54–fold; β6: −72, −122, −11, −3, −2000, −13, −338–fold).These results suggest that the β1, β3, and β6 conglutins have potential as functional food components in nutraceuticals and that can provide alternative therapies for the prevention and treatment of inflammatory-related diseases.

Effect of sucrose concentration and pH onto the physical stability of β-carotene nanocapsules

The incorporation of β-carotene into foods has a great impact due to its important biological properties and characteristic orange color that is of great interest for applications that require a natural colorant. However, it has limitations because of its lipophilic nature. Given this problem, an effective alternative is incorporating it as a functional ingredient in nanoencapsulation form, since this presents a significant benefit compared to emulsions/nanoemulsions; namely, increased solubility in aqueous media. In this study, the physical stability of dispersions was evaluated in relation to three factors: pH, sucrose concentration and different concentrations of β-carotene nanocapsules. Samples were characterized in terms of morphology, particle size, zeta potential, colorimetry, turbidity, and diffuse reflectance. Results show that no significant aggregation was detected, as revealed by turbidity (τ∼cte) and diffuse reflectance (p > 0.05), but that significant changes in chromaticity did occur during the storage period (p < 0.05) in all samples analyzed. The particle size obtained during monitoring was always below 300 nm. Minimal changes in zeta potential in the samples was evaluated. Given that these β-carotene nanocapsules showed only minimal changes in physical stability during storage, they may represent an effective option as a component of functional beverages and foods, as well as nutraceutical products.

Preparation and characterisation of a novel hydrogel based on Auricularia polytricha β-glucan and its bio-release property for vitamin B12 delivery.

This study investigates a novel hydrogel synthesis method and its bio-release property. This hydrogel, with a three-dimensional network structure based on Auricularia polytricha β-glucan, was characterised by means of Fourier transform infrared spectroscopy, 1 H NMR and scanning electron microscopy. Vitamin B12 (VB12 , cobalamin) as a hydrophilic functional food component was entrapped into these hydrogels. The in vitro release profile of VB12 was established in simulated gastric fluid (SGF) and simulated intestinal fluid (SIF). The results showed that the hydrogel had medium pore size from 30 to 300 µm, and the swelling ratio increased with the degree of substitution. The hydrogel demonstrated good stability in SGF and bio-release capability in SIF for VB12 . The accumulated release rate is about 80% in SIF and below 20% in SGF, which indicated the significant different release property in stomach and intestine. The Auricularia polytricha β-glucan-based hydrogel has a good swelling ratio, pepsin stability and pancrelipase-catalysed biodegradation property. The bio-release rate is significantly different in SIF and SGF, which indicated that this hydrogel could be a good intestinal target carrier of VB12 . © 2017 Society of Chemical Industry.

Biological evaluation and quantitative analysis of antioxidant compounds in pulps of the Amazonian fruits bacuri (Platonia insignisMart.), ingá (Inga edulisMart.), and uchi (Sacoglottis uchiHuber) by UHPLC-ESI-MS/MS

Bacuri, inga, and uchi are Amazon fruits consumed specially in the North region of Brazil. Due to their large consumption and the lack of knowledge regarding its chemical composition, these fruits were studied in relation to their Antioxidant chemical constitution. The total phenolic content ranged from 3.86 ± 0.47 to 33.38 ± 1.51 mg GAE/100 g, and the total flavonoid content ranged from 1.75 ± 0.22 to 19.44 ± 0.87 mg QUERE/100 g, where the contents showed a significant correlation with DPPH and ABTS antioxidant assays. Thus, UHPLC-MS/MS was applied do quantify selected compounds, been citric acid the most abundant for all fruits. Furthermore, samples were screened for their α-glycosidase and lipase inhibitory effects, in addition to their antimicrobial potentials. Bacuri showed the highest antioxidant and α-glycosidase inhibitory capacity (IC50 15.20 μg/mL), whereas uchi and its main metabolite bergenin displayed moderate antimicrobial activities. The results shed light into the potentials of Amazonian fruit sources. Practical applications Plant phenolics are essential components of functional foods, due to their antioxidant and enzyme inhibition activities, which are directly linked to several diseases prevention. This is the first study about the quantification of antioxidant compounds in the Amazonian fruits: bacuri, inga, and uchi. Although they are quite consumed in the North region of Brazil, there are no bio-products made from them. This study aimed to elucidate the knowledge about the chemical composition and potentialities within these fruits with the practical purpose of highlighting them to future commercial applications. In addition, once we indicate their capabilities, we contribute with local populations in respect to the production of such fruits, which still is a family activity.

Potential health effects of the popular compound of artichoke: Cynarin (CYN)

Functional food components are usually related with disease prevention. Cynarin is one of the biologically active functional food components which is the major dicaffeoylquinic acid derivative found in artichoke. Although artichoke contains very low amount of cynarin, it still duplicates the effects of artichoke. Cynarin is one of the biologically active functional food components that have potential effects on choleretic and cholesterol lowering, hepatoprotective, anti-atherosclerotic, anti-HIV, antioxidative, anti-diabetic, anti-carcinogenic and immune modulator activity. To sum up, more molecular, clinical and epidemiological studies on cynarin is necessary to be able to use it as a disease preventive agent.

Polysaccharides from Kochia scoparia fruits protect mice from lipopolysaccharide-mediated acute lung injury by inhibiting neutrophil elastase

Acute lung injury (ALI) is a respiratory disease that is associated with neutrophil activation. Neutrophil elastase released from activated neutrophils has been reported to cause lung tissue damage and contribute to the inflammatory response in ALI. In this study, aqueous extracts of Kochia scoparia fruits, a traditional botanical medicine and food garnish in Asia, showed a selective inhibitory effect on human neutrophil elastase (IC50, 79.53±4.26μgmL−1). Using bioactivity-guided fractionation, the polysaccharide fraction (KSWP, molecular weight>300kDa) was identified, and its protective effects against ALI were investigated. The results showed that KSWP protected mice from lipopolysaccharide-induced ALI by reducing elastase activity, neutrophil infiltration, pro-inflammatory cytokines, and neutrophil NET formation. In addition, KSWP showed high safety parameters in toxicity tests. These results conclusively demonstrate the high potency of KSWP as a component of functional foods or botanical products in the prevention of ALI.

Molecular Mechanisms and Pathways as Targets for Cancer Prevention and Progression with Dietary Compounds.

A unique feature of bioactive food ingredients is their broad antioxidant function. Antioxidants having a wide spectrum of chemical structure and activity beyond basic nutrition; display different health benefits by the prevention and progression of chronic diseases. Functional food components are capable of enhancing the natural antioxidant defense system by scavenging reactive oxygen and nitrogen species, protecting and repairing DNA damage, as well as modulating the signal transduction pathways and gene expression. Major pathways affected by bioactive food ingredients include the pro-inflammatory pathways regulated by nuclear factor kappa B (NF-κB), as well as those associated with cytokines and chemokines. The present review summarizes the importance of plant bioactives and their roles in the regulation of inflammatory pathways. Bioactives influence several physiological processes such as gene expression, cell cycle regulation, cell proliferation, cell migration, etc., resulting in cancer prevention. Cancer initiation is associated with changes in metabolic pathways such as glucose metabolism, and the effect of bioactives in normalizing this process has been provided. Initiation and progression of inflammatory bowel diseases (IBD) which increase the chances of developing of colorectal cancers can be downregulated by plant bioactives. Several aspects of the potential roles of microRNAs and epigenetic modifications in the development of cancers have also been presented.

Prospects in the use of aptamers for characterizing the structure and stability of bioactive proteins and peptides in food.

Food-derived bioactive proteins and peptides have gained acceptance among researchers, food manufacturers and consumers as health-enhancing functional food components that also serve as natural alternatives for disease prevention and/or management. Bioactivity in food proteins and peptides is determined by their conformations and binding characteristics, which in turn depend on their primary and secondary structures. To maintain their bioactivities, the molecular integrity of bioactive peptides must remain intact, and this warrants the study of peptide form and structure, ideally with robust, highly specific and sensitive techniques. Short single-stranded nucleic acids (i.e. aptamers) are known to have high affinity for cognate targets such as proteins and peptides. Aptamers can be produced cost-effectively and chemically derivatized to increase their stability and shelf life. Their improved binding characteristics and minimal modification of the target molecular signature suggests their suitability for real-time detection of conformational changes in both proteins and peptides. This review discusses the developmental progress of systematic evolution of ligands by exponential enrichment (SELEX), an iterative technology for generating cost-effective aptamers with low dissociation constants (K d) for monitoring the form and structure of bioactive proteins and peptides. The review also presents case studies of this technique in monitoring the structural stability of bioactive peptide formulations to encourage applications in functional foods. The challenges and potential of aptamers in this research field are also discussed. Graphical abstract Advancing bioactive proteins and peptide functionality via aptameric ligands.

Epigallocatechin gallate induces extracellular degradation of amyloid β-protein by increasing neprilysin secretion from astrocytes through activation of ERK and PI3K pathways

Amyloid-β (Aβ) production and clearance in the brain is a crucial focus of investigations into the pathogenesis of Alzheimer disease. Imbalance between production and clearance leads to accumulation of Aβ. The important Aβ-degrading enzymes in the brain are neprilysin (NEP) and insulin-degrading enzyme (IDE), and defective enzyme expression may facilitate Aβ deposition in sporadic late-onset AD patients. It has been suggested that epigallocatechin gallate (EGCG), a member of the catechin family, might be an effective treatment for AD, because it has been shown to elevate NEP expression. Therefore, we examined whether catechins, which are functional components of common foods, could regulate the degradation of Aβ by inducing NEP and IDE expression. We also investigated the role of catechins in activating intracellular signal transduction in astrocytes. Treatment of cultured rat astrocytes with EGCG significantly reduced the expression of NEP, but not IDE, in a concentration- and time-dependent manner. NEP expression in cultured astrocytes was suppressed by activation of extracellular signal-regulated kinase (ERK) and phosphoinositide 3-kinase (PI3K), and reduced NEP expression was accompanied by an increase of NEP release into the extracellular space (culture medium). Moreover, culture medium from EGCG-treated astrocytes facilitated the degradation of exogenous Aβ. These results suggest that EGCG may have a beneficial effect on AD by activating ERK-and PI3K-mediated pathways in astrocytes, thus increasing astrocyte secretion of NEP and facilitating degradation of Aβ.

Water properties in p\xe2t\xe9s enriched with potato juice

Freshly squeezed potato juice was used in the traditional European folk medicine. Potato juice can be introduced into food products both in its fresh as well as in thermally preserved form as a component of functional food. An attempt was made to analyse the impact of potato juice supplementation on qualitative and quantitative parameters characterising water bonds in poultry meat batters and pâtés. On the basis of temperature studies of water activity changes and spin–lattice and spin–spin relaxation times, the authors determined energy values of water transport in meat batters. It was found that supplementation with fresh potato juice limited significantly water transport. The performed analyses of temperature value changes of T2 spin–spin relaxation times revealed significant differences in the molecular dynamics of both water fractions in temperatures exceeding 50 °C. Measurement results of relaxation times in pâtés demonstrated that the amount of free water in relation to the amount of bound water in the examined systems depended on the addition of the probiotic factor and on the form in which it was added.

Transgenic silkworms secrete the recombinant glycosylated MRJP1 protein of Chinese honeybee, Apis cerana cerana.

Major royal jelly protein-1 (MRJP1) is the most abundant glycoprotein of royal jelly (RJ) and is considered a potential component of functional foods. In this study, we used silkworm transgenic technology to obtain five transgenic silkworm lineages expressing the exogenous recombinant Chinese honeybee, Apis cerana cerana, protein-1 (rAccMRJP1) under the control of a fibroin light chain (Fib-L) promoter in the posterior silk glands. The protein was successfully secreted into cocoons; specifically, the highest rAccMRJP1 protein content was 0.78% of the dried cocoons. Our results confirmed that the protein band of the exogenous rAccMRJP1 protein expressed in the transgenic silkworm lineages was a glycosylated protein. Therefore, this rAccMRJP1 protein could be used as an alternative standard protein sample to measure the freshness of RJ. Moreover, we also found that the overall trend between the expression of the endogenous and exogenous genes was that the expression level of the endogenous Fib-L gene declined as the expression of the exogenous rAccMRJP1 gene increased in the transgenic silkworm lineages. Thus, by employing genome editing technology to reduce silk protein expression levels, a silkworm bioreactor expression system could be developed as a highly successful system for producing various valuable heterologous proteins, potentially broadening the applications of the silkworm.

腸管上皮トランスポーターと機能性食品成分の相互作用( 第66回大会シンポジウム「ビタミン・バイオファクタートランスポーター研究の最前線」)

腸管上皮トランスポーターと機能性食品成分の相互作用( 第66回大会シンポジウム「ビタミン・バイオファクタートランスポーター研究の最前線」)

Anti-proliferative effect and cell cycle arrest induced by saponins extracted from tea (Camellia sinensis) flower in human ovarian cancer cells

Tea (Camellia sinensis) flower saponins (TFS) have various biological properties. However, the anti-cancer effects of TFS have not been investigated in any detail. Here, we evaluated the anti-cancer effects of TFS using human ovarian cancer cell lines. TFS (1.5μg/ml) produced significant antiproliferative effects against A2780/CP70 and OVCAR-3 cells by inducing p53-dependent apoptosis and S phase arrest. Further study showed that TFS decreased mitochondrial membrane potential, activated Caspase-3/7, Caspase-8 and Caspase-9 activities, and that the p53 inhibitor PFT-α reversed the TFS-induced cell growth inhibition and apoptosis. In addition, TFS inhibited the expression of Cdc25A, Cdk2, and CyclinD1 and upregulated Cyclin E and Cyclin A, suggesting that the Cdc25A-Cdk2-Cyclin E/A pathway was involved in TFS-induced S phase arrest. Furthermore, the S phase arrest was associated with a Chk2-Cdc25A DNA damage response. These results demonstrated that TFS has promising potential serving as functional food components for prevention of ovarian cancer.

Effects of melanin from Sepiella Maindroni ink (MSMI) on the intestinal Microbiome of mice

BackgroundBy the search for new natural compounds with beneficial health effects, cephalopod ink has been considered as an attempt to develop new drugs and functional foods, which is an especially active field in Asia, where cephalopods are a major fishery catch, for which ink sacs are a bi-product and where homeopathic medicine has deep roots. There is a demand to evaluate the safety and influence to the organism. The specific composition and relative abundance of the gut microbiota, which is potentially a major modulator of host metabolism, drives the interaction between functional foods and host health. We explore the effects of melanin from Sepiella Maindroni, most common cuttlefish in China, on the intestinal microbiome of mice.ResultsICR mice were randomly divided four groups, which were normal group (S), low melanin dose group (D; 120 mg/kg), medium melanin dose group (Z; 240 mg/kg), and high melanin dose group (G; 480 mg/kg). Melanin was delivered for 28 consecutive days. Fecal samples were used to generate 7715 operational taxonomic units (OTUs) via high-throughput sequencing. There were significant shifts in relative abundance of the dominant taxa at the phylum, class, order, family, and genus levels following melanin treatment.ConclusionsMSMI had no significant effect on the structure of intestinal flora in mice. The main effect was in the proportion of dominant bacterial communities. The effect positively correlated with the dose. From a health point of view, the use of melanin does not cause intestinal flora disorder. Our results may have important implications for MSMI as functional food component and potential therapeutic for manipulating gut microbiota.

Characterization and Immunomodulatory Activity of a Novel Peptide, ECFSTA, from Wheat Germ Globulin.

A novel peptide was extracted from wheat germ globulin and purified using ion-exchange chromatography, gel filtration chromatography, and semi-preparative reversed-phase high-performance liquid chromatography (RP-HPLC). The sequence of the peptide was found to be Glu-Cys-Phe-Ser-Thr-Ala (ECFSTA). Its immunomodulatory effects were evaluated, and the results showed that ECFSTA could enhance phagocytosis of RAW 264.7 cells and significantly increase their secretion of nitric oxide (NO), interleukin 6 (IL-6), tumor necrosis factor α (TNF-α), and reactive oxygen species (ROS). ECFSTA activated macrophages mainly through the pattern recognition receptors (PRRs) of toll-like receptor 2 (TLR2) and toll-like receptor 4 (TLR4), and the production of ROS simultaneously stimulated macrophages to produce TNF-α. Thus, ECFSTA could be used as an immunomodulator and might be a promising component of functional foods.

GABA promotes elastin synthesis and elastin fiber formation in normal human dermal fibroblasts (HDFs).

The multiple physiological effects of γ-aminobutyric acid (GABA) as a functional food component have been recently reported. We previously reported that GABA upregulated the expression of type I collagen in human dermal fibroblasts (HDFs), and that oral administration of GABA significantly increased skin elasticity. However, details of the regulatory mechanism still remain unknown. In this study, we further examined the effects of GABA on elastin synthesis and elastin fiber formation in HDFs. Real-time PCR indicated that GABA significantly increased the expression of tropoelastin transcript in a dose-dependent manner. Additionally, the expression of fibrillin-1, fibrillin-2, and fibulin-5/DANCE, but not lysyl oxidase and latent transforming factor-β-binding protein 4, were also significantly increased in HDFs. Finally, immunohistochemical analysis confirmed that treatment with GABA dramatically increased the formation of elastic fibers in HDFs. Taken together, our results showed that GABA improves skin elasticity in HDFs by upregulating elastin synthesis and elastin fiber formation.

Enhancing fructooligosaccharides production by genetic improvement of the industrial fungus Aspergillus niger ATCC 20611

Aspergillus niger ATCC20611 is one of the most potent filamentous fungi used commercially for production of fructooligosaccharides (FOS), which are prospective components of functional food by stimulating probiotic bacteria in the human gut. However, current strategies for improving FOS yield still rely on production process development. The genetic engineering approach hasn’t been applied in industrial strains to increase FOS production level. Here, an optimized polyethylene glycol (PEG)-mediated protoplast transformation system was established in A. niger ATCC 20611 and used for further strain improvement. The pyrithiamine resistance gene (ptrA) was selected as a dominant marker and protoplasts were prepared with high concentration (up to 108g−1 wet weight mycelium) by using mixed cell wall-lysing enzymes. The transformation frequency with ptrA can reach 30–50 transformants per μg of DNA. In addition, the efficiency of co-transformation with the EGFP reporter gene (egfp) was high (approx. 82%). Furthermore, an activity-improved variant of β-fructofuranosidase, FopA(A178P), was successfully overexpressed in A. niger ATCC 20611 by using the transformation system. The transformant, CM6, exhibited a 58% increase in specific β-fructofuranosidase activity (up to 507U/g), compared to the parental strain (320U/g), and effectively reduced the time needed for completion of FOS synthesis. These results illustrate the feasibility of strain improvement through genetic engineering for further enhancement of FOS production level.

Production of Manooligomannan from Palm Kernel Cake by Mannanase Produced from Streptomyces Cyaenus

&lt;p&gt;The increase of public attention to health has prompted researchers to look for new sources of functional food. Palm Cake Kernel (PKC) waste was abundant in Indonesia, Oligosaccharide has an important benefit for human health. Recently oligosaccharide is not only important as an artificial sweetener, but also as a functional food component. This study was aimed to produce oligo-mannan enzymatically from PKC waste using mannanase derived from of Streptomyces cyaenus isolates of indigenous Indonesia. The enzyme concentration was determined by enzyme activity assay while oligo-mannan content in the PKC was analyzed using TLC and HPLC. Mannanase enzyme activity of 1706 U/ml on the second day of agitation 200 rpm at a temperature of 30°C Hydrolysis of mannooligomannan by using mannanase produced by streptomyces cyaenus. The optimum mannanase enzyme activity obtained on day 2 with the value of the activity as much of 0.702 U/mL. The protein content of the 2nd day at an agitation speed of 150 rpm, 200 rpm, and 250 rpm, respectively, were 1783, 1950 and 2283 ppm. &lt;em&gt;Streptomyces cyaenus&lt;/em&gt; is Indonesian original isolates potentially producing mannanase that can produce mannooligomannan.&lt;/p&gt;

In vitro evaluation of whole faba bean and its seed coat as a potential source of functional food components.

In vitro studies were conducted to evaluate the particular nutritional benefits of whole faba bean seed (WFB) and fava bean seed coat (FBSC). Total dietary fiber contents of WFB and FBSC were 27.5% and 82.3%, respectively. FBSC were contained much higher total phenolic substances, condensed tannins, and total antioxidant activity than WFB. Bile acid (BA)-binding capacities of in vitro digested samples and nutritionally important products produced by in vitro fermentation of digestion residues were also studied. The BA-binding capacities of WFB and FBSC were 1.94 and 37.50μmol/100mg, respectively. Total BA bound by FBSC was even higher than the positive standard cholestyramine. Lignin and other constituents of the Klason residue were found to influence BA-binding properties. Moreover, the extent of the in vitro fermentation process showed that, fermentability of FBSC residue was significantly lower than that of WFB residue. Overall, faba bean, especially its seed coat, has great potential as a functional food.

Effect of luteolin on inflammatory responses in RAW264.7 macrophages activated with LPS and IFN-γ

Luteolin (Lut), a natural flavonoid extracted from plants, possesses several beneficial bioactivities. However, little is known about its anti-inflammatory properties. This study was designed to investigate the anti-inflammatory effects and underlying molecular mechanisms of luteolin in LPS plus IFN-γ activated RAW264.7 macrophages (M1 macrophages). RAW264.7 cells were induced to M1 polarization by LPS plus IFN-γ, accompanied with luteolin at non-cytotoxic doses. The results showed that the M1 macrophages presented dendrites characteristic withsynapsis. In addition, luteolin can inhibit inflammation by modulating the expression of mediators, such as iNOS, IL-1β, IL-6, TNF-α, and CD86, whose expression was increased remarkably in the activated RAW264.7 cells. More importantly, the proteins p-STAT3-ser727 and p-STAT3-tyr705, which involved in inflammatory pathways, exhibited a dose-dependent degradation in M1 macrophages after treatment with luteolin. In conclusion, luteolin have potential applications as a functional food component in regulating inflammatory responses.