Effects of Oral Supplementation with Paprika Xanthophylls on Human Skin Moisture.

Carotenoids, classified into carotenes and xanthophylls, are natural lipophilic pigments that are widely distributed in plants. Red paprika is unique in its high levels of various xanthophylls. Dietary paprika xanthophylls have been shown to reduce UV-induced photo damage by the strong antioxidant activity in the skin. However, the precise effects of paprika xanthophylls on skin condition are still unknown. Here we show that skin moisture is enhanced by the intake of red paprika supplements including seven xanthophylls. We conducted a 4-week randomized, single-blind, parallel-group controlled trial to clarify the effects of dietary paprika xanthophylls on facial skin. The results showed that the moisture was significantly higher in the paprika intake group than in the control (21.0±8.9 vs 13.4±6.0 (A.U.)). There was no significant difference between the paprika and control groups in other parameters such as viscoelasticity, the number of wrinkles, and the amount of water evaporation. On the other hand, the number of brown stains in the paprika group increased significantly, to 190±26 from 173±30 (p < 0.05). In vitro experiments, quantitative real-time PCR showed that paprika extract led to increases in the gene expression of Aquaporin 3 (AQP3) and hyaluronic acid synthase 3 (HAS3) in cultured keratinocytes. Western blotting showed that the paprika extract enhanced AQP3 expression. Taken together, taking supplements containing paprika xanthophylls may provide beneficial effects of moisture on facial skin. The study provides new insights into understanding the role of paprika xanthophylls in the skin.

Analysis of Fatty Acid Esters of Hydroxyl Fatty Acid in Nut Oils and Other Plant Oils.

Oils and lipids are common food components and efficient sources of energy. Both the quantity and the quality of oils and lipids are important with regard to health and disease. Fatty acid ester of hydroxy fatty acid (FAHFA) is a novel lipid class that was discovered as an endogenous lipid; FAHFAs have shown anti-diabetic effects in a mammalian system. We analyzed the overall FAHFA composition in nut oils and other common oils: almond (raw, roasted), walnut, peanut, olive, palm, soybean, and rapeseed oils. We developed a method of liquid chromatography coupled with electrospray ionization triple quadrupole mass spectrometry (LC-ESI/MS/MS) for a comprehensive target analysis of FAHFAs. The analysis revealed wide variation in the FAHFA profiles (15 compounds and 62 peaks). For 7-11 compounds of FAHFA, a total level of 8-29 pmol/mg oil was detected in nuts oils; for 11 compounds, 4.9 pmol/mg oil was detected in olive oil, and for 4-9 compounds, < 2 pmol/mg oil was detected in palm, soy, and rapeseed oils. The major FAHFAs were FAHFA 36:3, FAHFA 36:2, and FAHFA 36:4 in nut oil, FAHFA 36:2, FAHFA 34:1, and FAHFA 36:1 in olive oil, and FAHFA 32:1, FAHFA 34:0, FAHFA 36:0, and FAHFA 36:1 in all of the common oils. The composition of FAHFAs in nut oils is mainly unsaturated fatty acids, whereas those in olive oil are unsaturated fatty acids and saturated fatty acids. The composition of FAHFAs in common oils was mainly saturated fats. This is the first report to demonstrate the quality and quantity of the FAHFAs in the nut oils. Nuts have been described to be a great source of many nutrients and to be beneficial for our health. Our present findings comprise additional evidence that the intake of nuts in daily diets may prevent metabolic and inflammatory-based diseases.

Effects of Glycogen on Ceramide Production in Cultured Human Keratinocytes via Acid Sphingomyelinase Activation.

Glycogen is a highly branched storage polysaccharide found mainly in the liver and the muscles. Glycogen is also present in the skin, but its functional role is poorly understood. Recently, it has been reported that glycogen plays an important role in intracellular signal transduction. In the epidermis of the skin, keratinocytes are the predominant cells that produce ceramide. Ceramides are lipids composed of sphingosine, and prevent water loss, as well as protecting the skin against environmental stressors. In this study, we investigated the effects of glycogen on ceramide production in cultured keratinocytes. Thin-layer chromatography revealed that incubation of keratinocytes with 2 % glycogen enhanced the cellular amount of ceramide NS (ceramide 2) by 3.4-fold compared to the control. We also found that glycogen regulated the mRNA expression levels of signaling molecules of the sphingomyelin-ceramide pathway by quantitative real-time PCR. The activity of sphingomyelinase was also significantly enhanced by 2.5-fold in cultures with 1 % glycogen compared to the control. Moreover, glycogen increased the ATP production by 1.5-fold compared to the control, while glucose did not affect the production. Western blotting showed that phosphorylation of Akt, a cellular signaling molecule, was inhibited in the presence of glycogen in cultured keratinocytes. This study shows that glycogen upregulates the ceramide production pathway from sphingomyelin in epidermal keratinocytes, and provides new insights into the role of glycogen in cellular signal transduction.

Oiling-out effect improves the efficiency of extracting aroma compounds from edible oil.

Volatile compounds in foods are a significant factor that affects food intake and preference. However, volatile components in edible oils are poorly understood due to a strong matrix effect. In this study, we developed a method of extracting volatile compounds from extra virgin coconut oil (EVCO) by means of oiling-out assisted liquid-liquid extraction (OA-LLE). Consequently, 44 aroma compounds were isolated and identified from only 5 g of EVCO. Various aroma compounds were detected in addition to δ-lactones. The ratio of the natural abundance of the enantiomers of δ-lactones in EVCO was also revealed. Compared with the conventional methods of solvent assisted flavor evaporation (SAFE) and head-space solid-phase micro extraction (HS-SPME), OA-LLE was able to isolate a wide range and large number of volatile compounds from EVCO without leaving oil residues. Therefore, isolating aroma compounds from edible oil based on the oiling-out effect should provide an innovative extraction method.

Effects of Phosphoryl Oligosaccharides of Calcium (POs-Ca) on Mycelial Growth and Fruiting Body Development of the Edible Mushroom, Pleurotus ostreatus.

Phosphoryl oligosaccharides of calcium (POs-Ca) is a calcium salt of phosphoryl maltooligosaccharides made from potato starch. POs-Ca is highly water-soluble and can supply both the calcium ion and acidic oligosaccharides in an aqueous solution. In this study, we investigated the effects of POs-Ca on the mycelial growth and fruiting body yield of Pleurotus ostreatus , which is one of the most widely cultivated edible mushrooms in the world. We cultivated the mushroom using both potato dextrose agar (PDA) medium and sawdust-based medium, with added calcium salts. The addition of POs-Ca into the PDA medium with a calcium concentration of 10 mg increased mycelial growth significantly ( p < 0.05, vs . control). POs-Ca addition to the sawdust-based medium at concentrations of 1.0 to 3.0 g/100 g medium increased the amount of calcium in the fruiting bodies but did not affect the length of the cultivation period or the weight of the fruiting body. The calcium content in the fruiting body increased 12-fold when compared to the control. On the other hand, neither the CaHPO 4 ï½¥2H 2 O group nor the CaHPO 4 ï½¥2H 2 O with oligosaccharides group showed changes in the calcium content of the fruiting bodies. Our results indicate that the use of POs-Ca in mushroom cultivation allows for the possibility of developing new functional foods like calcium-enriched edible mushrooms. This is the first report describing the effects of POs-Ca on mushroom cultivation.

Co-ingestion of whole almonds and almond oil with carbohydrate suppresses postprandial glycaemia in mice in an insulin-dependent and insulin-independent manner.

Co-ingestion of almonds with carbohydrate prevents excessive increase in plasma glucose level (PGL), but information about the functional fraction is limited. Identifying the functional fraction is necessary to use almonds more efficiently in terms of controlling postprandial glycaemia after a high-carbohydrate meal. In the present study, we evaluated the effects of almond skin, oil, water-soluble fraction and water-insoluble fraction on both postprandial glycaemia and insulinaemia. The effect of almond skin was tested by comparing the effect of whole almonds with the effect of skinless almonds. Male ICR mice were administered dextrin and 4 g/kg body weight test samples. After the administration, 2-h postprandial changes in glycaemia and insulinaemia were measured. Oil was the only fraction being able to blunt postprandial glycaemia. Interestingly, when co-ingesting with dextrin, almond oil did not change the insulin level compared with the control but whole almonds or skinless almonds triggered a 4-fold increase in insulin level. The co-ingestion of whole almonds or skinless almonds similarly suppressed the PGL at 15 and 30 min (P < 0·05), which means almond skin has no effect on postprandial glycaemia. Neither soluble nor insoluble fractions lead to any significant changes in postprandial glycaemia and insulinaemia. In conclusion, oil is the main functional component accounting for the glycaemia-lowering effect without altering insulin level.

Effects of synbiotics containing Bifidobacterium animalis subsp. lactis GCL2505 and inulin on intestinal bifidobacteria: A randomized, placebo-controlled, crossover study.

A number of studies have shown the bifidogenic effects of either probiotic bifidobacteria or inulin, and this bifidogenic shift in the composition of the colonic microbiota is likely the basis for their positive impact on human health. This study aimed to evaluate the effects of synbiotics containing the probiotic bacterium Bifidobacterium animalis subsp. lactis (B. lactis) GCL2505 and inulin on the levels of intestinal bifidobacteria compared with B. lactis GCL2505 alone. A randomized, double-blind, placebo-controlled, crossover trial was carried out involving 60 healthy subjects with a tendency for constipation using fermented milk containing B. lactis GCL2505 and inulin (synbiotic), only B. lactis GCL2505 (probiotic), and placebo. Fecal samples were collected at the end of each 2-week intervention period, and the bifidobacterial count was analyzed by quantitative real-time PCR. The numbers of total bifidobacteria and B. lactis in feces were significantly increased during the probiotic and synbiotic intake periods compared with the placebo intake period. Furthermore, the numbers of total bifidobacteria and endogenous bifidobacteria were significantly higher in the synbiotic intake period compared with the probiotic intake period, while there was no difference in the number of B. lactis. These results suggested that the synbiotics containing B. lactis GCL2505 and inulin had a greater effect on the number of bifidobacteria than a drink containing probiotics alone and could be useful for the improvement of the intestinal environment.

Phosphoryl oligosaccharides of calcium enhance mineral availability and fluorapatite formation.

OBJECTIVES: Phosphoryl oligosaccharides of calcium (POs-Ca) are a highly soluble calcium source and can keep the solubility of calcium and fluoride ions. The aim of this study was to investigate the effect of calcium (from POs-Ca) and fluoride ions penetrate into subsurface enamel lesions in vitro. DESIGN: Demineralized bovine enamel slabs were remineralizedin vitro for 24 h at 37 °C with artificial saliva (AS) containing POs-Ca and various fluoride concentrations (0-100 ppm), or AS containing different levels of POs-Ca adjusted to a Ca/P ratio of 0.4-3.0 and fluoride, then were analyzed using Transversal microradiography. From those results, remineralization effects with optimal conditions were compared between POs-Ca and calcium chloride (CaCl2). To determine the form of incorporated fluoride, we analyzed the chemical state and local structure of fluorine atoms integrated into enamel subsurface lesions using micro X-ray absorption near-edge structure (µ-XANES) spectroscopy. RESULTS: A significant mineral recovery rate was observed with POs-Ca and fluoride at 0.5 or 1.0 ppm (n = 6, p < 0.05), as well as a Ca/P molar ratio of 1.67 (n = 5, p < 0.05). Under those conditions, the mineral recovery rate of AS containing POs-Ca (37.9 ± 7.3%) was significantly greater than that of CaCl2 (15.0 ± 9.6%) (n = 5, mean ± SD, p < 0.05). µ-XANES spectra analysis of the samples indicated that the dominant form of fluorine atoms in enamel subsurface lesions was fluorapatite. CONCLUSIONS: POs-Ca with fluoride-derived diffusion into subsurface enamel lesions facilitated formation of fluorapatite phases.

Predicting organ carotenoids levels from analysis of plasma could lead to errors: A study in cynomolgus monkeys.

Carotenoids are phytochemicals with strong antioxidant activity against reactive oxygen species that are widely distributed in fruits and vegetables. The beneficial effects of carotenoids on human health have attracted considerable attention. The plasma carotenoid profile in humans is generally recognized to reflect the dietary carotenoid composition. Although carotenoid profile in plasma is believed to correlate well with that in other tissues, the data for tissue accumulation of carotenoids in humans is very limited and poorly understood. In order to test the hypothesis that blood carotenoids reflect tissue accumulation of dietary carotenoids, the cynomolgus monkey was used as a model to determine it's suitable for extrapolation of data on tissue accumulation of carotenoids to humans. Herein, plasma carotenoids were measured in cynomolgus monkeys given a dietary mixture of carotenoids. The findings indicate that cynomolgus monkeys and humans are similar with regard to preferential accumulation of ß-cryptoxanthin in the blood and brain. These results suggested that cynomolgus monkeys could be used to collect data on tissue accumulation of carotenoids for extrapolation to humans. The tissue accumulation of carotenoids in other tissues of cynomolgus monkeys that have not yet been evaluated in humans were also investigated, revealing marked differences in carotenoid levels and composition among plasma and various monkey tissues. These results suggest that accumulation of carotenoids in plasma does not reflect necessarily that in tissues, so that predicting the tissue accumulation of carotenoids from plasma carotenoid levels and profiles alone could lead to errors.

Effect of Oral Paprika Xanthophyll Intake on Abdominal Fat in Healthy Overweight Humans: A Randomized, Double-blind, Placebo-controlled Study.

PURPOSE: Xanthophylls that exist in various vegetables and fruits have beneficial actions, such as antioxidant activity and an anti-metabolic syndrome effect, and daily intake of xanthophylls could play an important role in preventing lifestyle-related diseases. We investigated whether intake of xanthophylls from red paprika could decrease the abdominal fat area in the healthy overweight volunteers with a body mass index (BMI) ranging from 25 to < 30 kg/m2. METHODS: In a randomized, double-blind, placebo-controlled, parallel-group study, 100 healthy volunteers were assigned to oral administration of paprika xanthophyll capsules (containing 9.0 mg of paprika xanthophylls) or placebo capsules for 12 weeks. The primary endpoint was the effect of paprika xanthophyll intake on the abdominal visceral fat area (VFA) as determined by computed tomography. The secondary endpoints were as follows: 1) changes of the abdominal subcutaneous fat area (SFA), total fat area (TFA), and BMI; 2) changes of lipid metabolism parameters, glucose metabolism parameters, and other blood parameters. RESULTS: After 12 weeks, VFA was smaller in the paprika xanthophyll group than in the placebo group. In the paprika xanthophyll group, there was a significant decrease of SFA, TFA, and BMI after 12 weeks compared with baseline, and the reduction of SFA, TFA, and BMI was significantly greater in the paprika xanthophyll group than in the placebo group. Moreover, total cholesterol and low-density lipoprotein cholesterol decreased significantly in the paprika xanthophyll group, but not in the placebo group. No adverse effects were caused by intake of paprika xanthophyll capsules. CONCLUSIONS: Intake of paprika xanthophylls for 12 weeks significantly reduced the abdominal fat area and BMI in healthy overweight volunteers without causing any adverse effects. These findings suggest that paprika xanthophyll is a safe food ingredient that improves lipid metabolism and reduces abdominal fat. TRIAL REGISTRATION: UMIN-CTR UMIN000021529.

Effects of Dietary Paprika Xanthophylls on Ultraviolet Light-Induced Skin Damage: A Double-Blind Placebo-Controlled Study.

Generation of singlet oxygen by solar ultraviolet (UV) radiation causes acute inflammatory responses in the skin. Accumulation of singlet-oxygen-quenching antioxidants in the skin can suppress this photo-oxidative stress. This study evaluated the effect of dietary xanthophylls from red paprika fruit extract on UV-induced skin damage. A randomised double-blind placebo-controlled parallel group comparison study involving 46 healthy volunteers was performed. The minimal erythema dose (MED) of each individual was determined prior to the study. A capsule containing paprika xanthophylls (9 mg) or a placebo was administered daily for 5 weeks. The MED, minimal tanning dose (MTD), skin physiology parameters (skin color, hydration, and barrier function), and facial skin physiology parameters were evaluated at weeks 0, 2, and 4. The MED of the verum group at 2 and 4 weeks after administration was significantly higher than that of the placebo group. At 4 weeks, the suppression of UV-induced skin darkening by the verum diet was significantly greater than that of the placebo. There were no significant differences in facial skin parameters between the verum and placebo groups. Our results indicate the efficacy of dietary paprika xanthophylls in suppression of UV-induced skin damage.

Qualitative and Quantitative Analyses of Glycogen in Human Milk.

Identification as well as a detailed analysis of glycogen in human milk has not been shown yet. The present study confirmed that glycogen is contained in human milk by qualitative and quantitative analyses. High-performance anion exchange chromatography (HPAEC) and high-performance size exclusion chromatography with a multiangle laser light scattering detector (HPSEC-MALLS) were used for qualitative analysis of glycogen in human milk. Quantitative analysis was carried out by using samples obtained from the individual milks. The result revealed that the concentration of human milk glycogen varied depending on the mother's condition-such as the period postpartum and inflammation. The amounts of glycogen in human milk collected at 0 and 1-2 months postpartum were higher than in milk collected at 3-14 months postpartum. In the milk from mothers with severe mastitis, the concentration of glycogen was about 40 times higher than that in normal milk.

Accumulation of Paprika Carotenoids in Human Plasma and Erythrocytes.

The accumulation (incorporation) of paprika carotenoid in human plasma and erythrocytes was investigated. A paprika carotenoid supplement (14 mg/day) was ingested for 4 weeks by 5 young healthy volunteers (3 men and 2 women). After 2 weeks of carotenoid ingestion, the carotenoid levels in plasma and erythrocytes increased by 1.2-fold and 2.2-fold, respectively. Characteristic carotenoids found in paprika (capsanthin, cucurbitaxanthin A, and cryptocapsin) were detected in both plasma and erythrocytes. An oxidative metabolite of capsanthin (capsanthone) was also found in both plasma and erythrocytes.

Effects of ingesting highly branched cyclic dextrin during endurance exercise on rating of perceived exertion and blood components associated with energy metabolism.

We compared the effect of relatively low doses (15 g) of highly branched cyclic dextrin (HBCD) with that of maltodextrin during endurance exercise on the rating of perceived exertion (RPE) in a crossover, double-blind study of healthy volunteers. The RPE increased during exercise and its increase was significantly less at 30 and 60 min after ingesting HBCD than maltodextrin.

Enzymatically synthesized glycogen reduces lipid accumulation in diet-induced obese rats.

Based on a recent study indicating that enzymatically synthesized glycogen (ESG) possesses a dietary, fiber-like action, we hypothesized that ESG can reduce the risk of obesity. In this study, the antiobesity effects of ESG were investigated in a model of diet-induced obesity. Male Sprague-Dawley rats were divided into 4 groups and fed a normal or high-fat diet, with or without 20% ESG, for 4 weeks. Body weight, food intake, lipid deposition in the white adipose tissues and liver, fecal lipid excretion, and plasma lipid profiles were measured. At week 3, the body fat mass was measured using an x-ray computed tomography system, which showed that ESG significantly suppressed the high-fat diet-induced lipid accumulation. Similar results were observed in the weight of the adipose tissue after the experiment. Moreover, ESG significantly suppressed the lipid accumulation in the liver but increased fecal lipid excretion. The plasma concentrations of triacylglycerol and nonesterified fatty acid were lowered after a high-fat diet, whereas the total bile acid concentration was increased by ESG. However, the hepatic messenger RNA (mRNA) levels of enzymes related to lipid metabolism were not affected by ESG. Conversely, the mRNA levels of long-chain acyl-CoA dehydrogenase and medium-chain acyl-CoA dehydrogenase were up-regulated by ESG in the muscle. These results suggest that the combined effects of increased fecal lipid excretion, increased mRNA levels of enzymes that oxidize fatty acids in the muscle, and increased total bile acid concentration in the plasma mediate the inhibitory effect of ESG on lipid accumulation.

Optimization of calcium concentration of saliva with phosphoryl oligosaccharides of calcium (POs-Ca) for enamel remineralization in vitro.

OBJECTIVE: Phosphoryl oligosaccharides of calcium (POs-Ca) are highly soluble calcium source made from potato starch. The aim of this study was to investigate the optimal concentrations of POs-Ca for the remineralization of subsurface enamel lesions in vitro. DESIGN: Demineralized bovine enamel slabs (n=5) were remineralized in vitro for 24h at 37°C with artificial saliva (AS) containing 0-0.74% POs-Ca to adjust the Ca/P ratio to 0.4-3.0, then sectioned and analysed by transversal microradiography (TMR). The data were analysed by Scheffe's post hoc test. The Ca/P ratio with most remineralization was used to investigate the effect of calcium on enamel remineralization (n=11). The demineralized slabs were treated with AS with calcium-chloride- (CaCl2-) or POs-Ca with an identical calcium content, and sectioned for TMR and wide-angle X-ray diffraction (WAXRD) analyses to evaluate the local changes in hydroxyapatite (HAp) crystal content. The data were analysed using the Mann-Whitney U-test. RESULTS: The highest mineral recovery rate resulted from addition of POs-Ca to adjust the Ca/P to 1.67. At this ratio, the mineral recovery rate for AS containing POs-Ca (24.2±7.4%) was significantly higher than that for AS containing CaCl2 (12.5±11.3%) (mean±SD, p<0.05). The recovery rate of HAp crystallites for AS containing POs-Ca (35.7±10.9%) was also significantly higher than that for AS containing CaCl2 (23.1±13.5%) (p<0.05). The restored crystallites were oriented in the same directions as in sound enamel. CONCLUSIONS: POs-Ca effectively enhances enamel remineralization with ordered HAp at a Ca/P ratio of 1.67.

The effect of orally administered glycogen on anti-tumor activity and natural killer cell activity in mice.

Natural killer (NK) cells, innate immune effectors that mediate rapid responses to various antigens, play an important role in potentiating host defenses through the clearance of tumor cells and virally infected cells. By using enzymatically synthesized glycogen (ESG) with the same characteristics as natural glycogen, we examined whether orally administered glycogen enhances the innate defense of tumor-implanted mice and the cytotoxicity of NK cells. Oral administration of ESG led to the suppression of tumor proliferation and the prolongation of survival times of tumor-bearing mice. Splenic NK activities of BALB/c mice treated orally with ESG were significantly higher than those of water-treated mice, which were used as a negative control. In addition, intraduodenal injections of ESG gradually and markedly lowered splenic sympathetic nerve activity, which has an inverse correlation with NK activity. Furthermore, ESG activated Peyer's patch cells to induce the production of macrophage inflammatory protein-2 (MIP-2), interleukin-6 (IL-6), and immunoglobulin A (IgA) from these cells. These results demonstrated that orally administrated glycogen significantly enhanced the cytotoxicity of NK cells by acting on Peyer's patch cells and autonomic nerves, and eventually induced the potentiation of host defenses. We propose that glycogen functions not only as an energy source for life support but also as an oral adjuvant for immunopotentiation.

Essential role of Toll-like receptor 2 in macrophage activation by glycogen.

We prepared enzymatically synthesized glycogen (ESG) with the same characteristics as natural glycogen and investigated whether the macrophage-stimulating activity of glycogen was related to Toll-like receptors (TLRs), which are important receptors for innate immunity. ESG induced no nuclear factor-kappa B (NF-κB) activity in TLR4/MD-2/CD14-expressed human embryonic kidney 293 (HEK293) reporter cells, whereas this polysaccharide did activate peritoneal exude cells (PECs) derived from TLR4-deficient mice at the same level as those from wild-type (WT) mice. Similarly, ESG did not activate HEK293 cells expressing TLR3, 5, 7, 8 or 9, suggesting that these TLRs were irrelevant to the activity of ESG. In contrast, ESG enhanced the NF-κB activity of TLR2-expressed HEK293 reporter cells in a concentration-dependent manner. Furthermore, the cell-stimulating activity of ESG was remarkably lower for PECs from TLR2-deficient mice compared with those from WT mice. The activity of ESG completely disappeared after treatment with a glycogen-degrading enzyme, indicating that the activity derived from ESG itself and not from contamination with canonical TLR2 ligands such as bacterial lipopeptides. Moreover, it was clarified by ELISA that ESG was directly bound to TLR2. Taken together, these results demonstrated that TLR2 directly recognizes glycogen and that the recognition activates immunocytes such as macrophages to enhance the production of nitric oxide and inflammatory cytokines. In addition, it was suggested that TLR2 could be involved in the glycogen activity in vivo. We propose that glycogen act as an activator to potentiate the host defense through TLR2 on the macrophage.

Metabolic fate of orally administered enzymatically synthesized glycogen in rats.

We developed a new process for enzymatically synthesized glycogen (ESG), which is equivalent in physicochemical properties to natural-source glycogen (NSG) except its resistant property to degradation by α-amylase in vitro. In this study the metabolic fates of orally administered ESG in rats were investigated by a single oral administration test and a 2 week ingestion test. The glycemic index of ESG was 79. After the 2 week ingestion of ESG, the cecal content and production of short chain fatty acids were significantly increased, the pH value of cecal content was lowered, and the counts of Bifidobacterium and Lactobacillus in feces were significantly increased. Additionally, plasma levels of triacylglycerol and total cholesterol were significantly reduced by ESG. In contrast, NSG did not affect these parameters at all. The results collectively suggest that around 20% of orally administered ESG was transferred to the cecum in the form of polymer and assimilated into short chain fatty acids by microbiota and the polymer affected lipid metabolism.

Structural basis for branching-enzyme activity of glycoside hydrolase family 57: structure and stability studies of a novel branching enzyme from the hyperthermophilic archaeon Thermococcus kodakaraensis KOD1.

Branching enzymes (BEs) catalyze the formation of branch points in glycogen and amylopectin by cleavage of α-1,4 glycosidic bonds and subsequent transfer to a new α-1,6 position. BEs generally belong to glycoside hydrolase family 13 (GH13); however TK1436, isolated from the hyperthermophilic archaeon Thermococcus kodakaraensis KOD1, is the first GH57 member, which possesses BE activity. To date, the only BE structure that had been determined is a GH13-type from Escherichia coli. Herein, we have determined the crystal structure of TK1436 in the native state and in complex with glucose and substrate mimetics that permitted mapping of the substrate-binding channel and identification of key residues for glucanotransferase activity. Its structure encompasses a distorted (ß/α)(7)-barrel juxtaposed to a C-terminal α-helical domain, which also participates in the formation of the active-site cleft. The active site comprises two acidic catalytic residues (Glu183 and Asp354), the polarizer His10, aromatic gate-keepers (Trp28, Trp270, Trp407, and Trp416) and the residue Tyr233, which is fully conserved among GH13- and GH57-type BEs. Despite TK1436 displaying a completely different fold and domain organization when compared to E. coli BE, they share the same structural determinants for BE activity. Structural comparison with AmyC, a GH57 α-amylase devoid of BE activity, revealed that the catalytic loop involved in substrate recognition and binding, is shortened in AmyC structure and it has been addressed as a key feature for its inability for glucanotransferase activity. The oligomerization has also been pointed out as a possible determinant for functional differentiation among GH57 members.