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1.
Using a combination of High‐Performance Ion Chromatography analysis and kinetic studies the stereospedfidty of myo‐inositol hexakisphosphate dephosphorylation by the spelt phytase D21, the rye phytase, the barley phytases P1 and P2, and the oat phytase was established. The data demonstrate that all the cereal phytate‐degrading enzymes under investigation dephosphorylate myo‐inositol hexakisphosphate in a stereospecific way by sequential removal of phosphate groups via D‐Ins(l,2, 3,5, 6)P5, D‐Ins(l,2, 5,6)P4, D‐Ins(l,2, 6)P3, D‐Ins(l,2)P2 to finally Ins(2)P (notation 4/3/5/6/1).  相似文献   

2.
Abstract: Two assays were conducted to investigate the changes of faba bean (Vicia faba L.) and azuki bean (Vigna angularis L.) phosphatases (phytase [Phy] and acid phosphatase [AcPh]) and the degradation of its substrates (inositol phosphate esters) during seed germination. The 1st assay was to establish the optimal germination conditions of faba bean and azuki bean to improve the endogenous phosphatases and increase the hydrolysis of phytate and, in the second assay, to determine the different lower phosphate esters of myo‐inositol produced during the germination process. In the 1st assay, seeds were soaked for 12 and 24 h and germinated for 3 and 5 d with and without the addition of gibberellic acid (GA3). In the second assay, seeds were soaked for 12 h and germinated for 1, 3, and 5 d with GA3. Phy (up to 3625 and 1340 U/kg) and AcPh (up to 9456 and 2740 U/g) activities, and inositol hexaphosphate (IP6) (8.23 and 7.46 mg/g), inositol pentaphosphate (IP5) (0.55 and 0.82 mg/g), and inositol tetraphosphate (IP4) (0.26 and 0.01 mg/g) were detected in ungerminated faba bean and azuki bean, respectively. The germination process caused a significant increase of Phy and AcPh activities in faba bean (up to 147% and 210%) and azuki bean (up to 211% and 596%) and a reduction in the phytate phosphorus content (up to 81% and 63%, respectively). Phytate phosphorus content was affected only by soaking time in the case of faba bean. Finally, during the course of germination, IP6 and IP5 were rapidly degraded in faba bean (88% and 39%) and azuki bean (55% and 56%), and IP4 was only a short‐living intermediate, which was increased during hydrolysis and degraded to inositol triphosphate. In this manner we could obtain a low‐phytate, endogenous phosphatase‐rich ingredient for enhancing human nutrition.  相似文献   

3.
The degradation and formation of inositol phosphates as affected by microbial phytase and gastrointestinal enzyme activities during the passage of phytate through the stomach and small intestine were studied in two experiments with four barrows and three collection periods. The degradation and formation of inositol phosphates were measured at the duodenal and ileal sites using Cr‐NDR, TiO2 and Co‐EDTA as indigestible markers. In experiment 1, the effect of graded doses of Aspergillus niger phytase (0, 150 and 900 FTU Natuphos® kg?1), added to a maize–soybean meal‐based diet with very low intrinsic phytase activity on the degradation of phytate and the formation of inositol phosphates during digestion in the stomach and small intestine was investigated. In experiment 2, three different mixtures of inositol phosphates, produced by Aspergillus niger phytase, containing mainly high, intermediate and low phosphorylated inositol phosphates, were added to the same maize–soybean meal‐based diet as used in experiment I. The fate of the inositol phosphates during digestion in the stomach and small intestine was studied. Experiment 1 showed that the extent of phytate degradation was dependent of the graded dietary phytase activities. At high phytase activity (900 FTU kg?1 of diet), strong phytate degradation occurred and the once hydrolysed phytate was rapidly dephosphorylated to lower inositol phosphates (mainly inositol di‐ and triphosphates). Intermediate inositol phosphates, such as inositol tetraphosphates, were quantitatively unimportant in duodenal and ileal digesta. At a phytase activity of 150 FTU kg?1 of diet, a broader spectrum of intermediate inositol phosphates was determined, which was probably due to a slower breakdown of phytate. Experiment 2 showed as a predominant result that lower inositol phosphates such InsP4 and InsP3 were degraded, whereas InsP2 accumulated in the duodenal and ileal digesta. No substantial disappearance of phytate from the stomach and small intestine was found when high concentrations of soluble phytate were added to the diet, which indicates that no substantial phytate absorption occurs in the upper part of the pig gut. Copyright © 2005 Society of Chemical Industry  相似文献   

4.
Scope: The objective of this study is to investigate a vascular effect of N‐(p‐coumaroyl)serotonin (CS) and N‐feruloylserotonin (FS), major antioxidative indolic polyphenols in safflower seeds with anti‐atherogenic properties, with emphasis on effects on vascular smooth muscle cells (VSMCs). Methods and results: Both CS and FS (each 10 to 100 μM) relaxed rat femoral arteries, which were pre‐contracted by 10?5 M phenylephrine or 50 mM KCl, independently of their endothelium. Both CS and FS also concentration‐dependently inhibited the increase of cytosolic free Ca2+ concentration ([Ca2+]i) that was induced by KCl or 5‐hydroxytryptamine in cultured rat VSMCs. Next, we examined the effects of CS and FS on platelet‐derived growth factor (PDGF)‐BB‐evoked proliferation and migration of the VSMCs. Both CS and FS inhibited PDGF‐BB‐evoked proliferation and migration of the VSMCs in a concentration‐dependent manner. They also inhibited PDGF‐BB‐induced phosphorylation of PDGF receptor β and ERK1/2, and Ca2+ release from sarcoplasmic reticulum in the VSMCs in a concentration‐dependent fashion. Conclusion: These results indicated a possible vascular effect of CS/FS to inhibit the activation of VSMCs by blocking the increase of [Ca2+]i and/or blocking PDGF signaling. These may explain a part of anti‐atherogenic mechanism that underlies their ability to improve vascular distensibility and to inhibit aortic hyperplasia.  相似文献   

5.
Phytate (inositol hexa‐phosphate or IP6) possessing anticancer activity is hydrolyzed by phytase in intestinal microbes and the metabolites are distributed throughout the colon. Cellular circumferential F‐actin rings, which are involved in cell polarity and structure, are lost early during tumorigenesis. We investigated F‐actin ring formation by the phytate hydrolysate in colorectal cancer HT‐29 cells to explore the novel mechanisms underlying the phytate‐mediated anticancer function. The phytate hydrolysate, but not inositol or phytate, induced F‐actin ring formation with a peak at 10 min in the cells and was associated with phosphorylation of myosin regulatory light chain. F‐actin ring formation and myosin regulatory light chain phosphorylation by the phytate hydrolysate were suppressed by inhibitors of Rho‐associated kinase (ROCK), Janus kinase (JAK), c‐Jun N‐terminal kinase (JNK), and protein kinase Cδ (PKCδ). Activation of ROCK and JAK, but not JNK or PKCδ, was observed at 10 min and/or earlier after stimulation with the phytate hydrolysate. Altogether, the phytate hydrolysate induces circumferential F‐actin ring formation through a ROCK‐dependent myosin II activation in the HT‐29 cells, which requires JAK activation and basal activities of JNK and PKC. Hydrolysis products of phytate in the intestine may contribute to anticancer function of phytate.  相似文献   

6.
Changes in phytic acid content, activity of phytase and α-amylase in rye breads were determined during rye bread making. The activity of phytase is highest in grain and flour whereas the activity in the sourdoughs is almost the half of the activity in the flour. The activity was unchanged in the dough after mixing and proofing. Degradation of phytic acid (IP6) into lower inositol phosphates and free phosphate is almost completed during the production of rye bread with long fermentation time whereas the degradation is less completed when whole grains are included in the recipe. In rye bread made from milled rye (DB00), 99% of IP6 is degraded and IP3 becomes the dominating inositol phosphate in this bread type presumably resulting in a high level of bioavailable minerals. In rye bread made with 30% grains (SB30), 94% of the IP6 content was reduced with IP4 and IP3 being the dominating lower inositol phosphates. In rye bread made with 50% whole grains (KB50) the degradation of IP6 was 82%, and the three inositol phosphates IP5, IP4 and IP3 were found in equal amount in this bread type. Due to significant amounts of phytic acid and remaining IP5, some of the minerals might not be available for human absorption after consumption of this bread type.  相似文献   

7.
8.
Three in vitro experiments using a rumen fluid buffer system were performed to investigate the effect of addition of 4 experimental phytases (Phy1, Phy2, Phy3, and Phy4) compared with no addition of phytase on feed inositol phosphate hydrolysis in wheat and rapeseed cake to determine which of the 4 phytases was most suitable under rumen-like conditions. The feedstuffs were incubated with a mixture of physiological buffer, ruminal fluid, and exogenous phytase at pH 6.2, after which the samples were incubated for different periods. Incubations were stopped using HCl, and the samples were analyzed for inositol phosphates via high performance ion chromatography. Addition of phytase (Phy1) resulted in enhanced degradation of myo-inositol hexakisphosphate (InsP6) in rapeseed cake, whereas addition of exogenous phytase did not improve the degradation of InsP6 in wheat. Only rapeseed cake was therefore used subsequently. All 4 phytases increased degradation of InsP6 in rapeseed cake in the in vitro system, and degradability of InsP6 increased with higher incubation time and higher phytase dosages, independent of phytase. Addition of 2 units of phytase per gram of substrate of the phytases Phy1, Phy2, Phy3, and Phy4 led to an undegraded InsP6 content of 56, 49, 70, and 18%, respectively, when incubated with rapeseed cake for 6 h, indicating that Phy2 and Phy4 were the most effective phytases. However, Phy2 had a higher specific activity than Phy4, as 60% of the original InsP6 content was remaining after 3 h when 5 mg of enzyme protein per gram of substrate of Phy2 was added to rapeseed cake, whereas 150 mg of enzyme protein per gram of substrate of Phy4 was necessary to achieve a similar result. Therefore, Phy2 appeared to be most applicable under rumen-like conditions.  相似文献   

9.
This study was designed to investigate the effects of Ca2+ and Zn2+ ions injection on post‐mortem tenderness and caspase activation by examining the expression of caspase‐3 and cytochrome c in post‐mortem beef skeletal muscles treated with Ca2+ and Zn2+. Results showed that Ca2+ ions injection accelerated post‐mortem tenderisation of beef skeletal muscles, whereas Zn2+ retarded the process. In the Ca2+‐injected samples, the level of caspase‐3 precursor decreased markedly without the production of activated caspase‐3, whereas the level of cytochrome c was increased markedly. So, Ca2+ possibly promoted caspases activation upstream of cytochrome c release, but inactivated caspase activity by calpain and/or fast depletion of ATP; whereas Zn2+ blocked the activation of procaspase‐3 with no visible change in the level of cytochrome c, and the block possibly resulted from its direct inhibition on caspase‐3 enzyme.  相似文献   

10.
Asp‐Leu‐Glu‐Glu (DLEE) is one of the antioxidant peptides purified from Chinese dry‐cured Xuanwei ham in our previous study. In the current work, the stability in a simulated digestion system, the transportation pathway and the antioxidant ability of DLEE were further investigated in a Caco‐2 cell monolayer. In the simulated gastrointestinal digestion system, no oligopeptides were generated. In the transport trial, the inhibitors cytochalasin D increased the transport of DLEE across the Caco‐2 cell monolayer, with Papp values of 3.22 × 10?6 cm s?1. A decreased expression occludin was observed with the DLEE incubation in the cell monolayer, and the antioxidant activity showed to be increased gradually in basolateral side. This study indicates that the absorption of DLEE could mainly occur via paracellular transport and provides information about its antioxidant activity after being absorbed across a cell monolayer.  相似文献   

11.
The pH buffering capacity of cheese is an important determinant of cheese pH. However, the effects of different constituents of cheese on its pH buffering capacity have not been fully clarified. The objective of this study was to characterize the chemical species and chemical equilibria that are responsible for the pH buffering properties of cheese. Eight cheeses with 2 levels of Ca and P (0.67 and 0.47% vs. 0.53 and 0.39%, respectively), residual lactose (2.4 vs. 0.78%), and salt-to-moisture ratio (6.4 vs. 4.8%) were manufactured. The pH-titration curves for these cheeses were obtained by titrating cheese:water (1:39 wt/wt) dispersions with 1 N HCl, and backtitrating with 1 N NaOH. To understand the role of different chemical equilibria and the respective chemical species in controlling the pH of cheese, pH buffering was modeled mathematically. The 36 chemical species that were found to be relevant for modeling can be classified as cations (Na+, Ca2+, Mg2+), anions (phosphate, citrate, lactate), protein-bound amino acids with a side-chain pKa in the range of 3 to 9 (glutamate, histidine, serine phosphate, aspartate), metal ion complexes (phosphate, citrate, and lactate complexes of Na+, Ca2+, and Mg2+), and calcium phosphate precipitates. A set of 36 corresponding equations was solved to give the concentrations of all chemical species as a function of pH, allowing the prediction of buffering curves. Changes in the calculated species concentrations allowed the identification of the chemical species and chemical equilibria that dominate the pH buffering properties of cheese in different pH ranges. The model indicates that pH buffering in the pH range from 4.5 to 5.5 is predominantly due to a precipitate of Ca and phosphate, and the protonation equilibrium involving the side chains of protein-bound glutamate. In the literature, the precipitate is often referred to as amorphous colloidal calcium phosphate. A comparison of experimental data and model predictions shows that the buffering properties of the precipitate can be explained, assuming that it consists of hydroxyapatite [Ca5(OH)(PO4)3] or Ca3(PO4)2. The pH buffering in the region from pH 3.5 to 4.5 is due to protonation of side-chain carboxylates of protein-bound glutamate, aspartate, and lactate, in order of decreasing significance. In addition, pH buffering between pH 5 to 8 in the backtitration results from the reprecipitation of calcium and phosphate either as CaHPO4 or Ca4H(PO4)3.  相似文献   

12.
Evidence has accumulated concerning the medicinal application of Nelumbo nucifera in the treatment of various diseases. Neferine, an alkaloid from N. nucifera was found to exert cytotoxicity on liver cancer cells HepG2 in a dose-dependent manner. We evaluated its anticancer potential by studying its effect on mitochondrial membrane potential, intracellular calcium levels [Ca2+]i, cell membrane integrity, apoptotic body formation and DNA fragmentation in cultured HepG2 cells. The reactive oxygen species level has been increased upon neferine treatment with concomitant decrease in reduced glutathione. Our data further indicate reduction of ΔψM and increased [Ca2+]i during apoptosis induction by neferine with increased expression of apoptotic proteins such as Bax, Bad, cleaved forms of caspase 3, caspase 9 and PARP, with the downregulation of anti-apoptotic protein Bcl2 in HepG2 cells. Moreover, the expressions of tumour suppressor proteins p53 and PTEN were upregulated along with the downregulation of P-Akt. In addition, expression levels of TNF-α, p38 and ERK1/2 MAP kinases were increased upon neferine treatment. These results imply that mitochondrial-mediated ROS generation induced by neferine leads to caspase-dependent apoptosis in HepG2 cells.  相似文献   

13.
《Food chemistry》2003,81(2):233-239
The kinetics of inositol phosphate degradation during the action of naturally occurring endogenous phytase for up to 90 min in pea and lentil flours has been studied, and compared with the addition of commercial phytase enzyme. In raw lentils IP6, IP5, IP4 and IP3 were present, whilst in peas only the presence of IP6 and IP5 was observed. Endogenous phytases were activated when legume flour was suspended in acidified water at pH 5.5 and 37 °C, and significant differences between lentils and peas were found. IP6 suffered a sharp reduction in lentils and peas (81–91 and 73–93%, respectively), this reduction being slightly more pronounced after the addition of commercial phytase. The content of IP5 decreased in lentils (48–69%), and increased in peas, except when commercial phytase acted for the first 30 min, after which a reduction was found (23%). The content of IP4 generally decreased in lentils, except when endogenous phytase acted for 30 min when an increase was observed. However, in peas, IP4 appeared in high concentrations up to 60 min by the action of both endogenous and exogenous phytases. The content of IP3, on the other hand, did not change greatly in lentils. In peas it was not detected after the action of endogenous phytase enzyme and it appeared in a large amount after the action of commercial phytase. In order to obtain legume flour with low IP6 and IP5 contents and notable IP4 and IP3 contents, the action of naturally endogenous phytase for 30 min in lentils is recommendable, as well as the addition of commercial phytase enzyme for 60 min in peas.  相似文献   

14.
Bread is a staple food in many countries and an important source of iron and zinc. The bioavailability of these minerals is generally low because of the content of phytic acid. Traditional Iranian breads were prepared with flours of different extraction rates, Sangak at 93% and Barbari at 82%. Breads were dephytinised by addition of Aspergillus niger phytase during in vitro digestion. The effect upon iron and zinc bioaccessibility in the Caco‐2 cell model was investigated. The cellular uptake of iron and zinc was lower from Sangak, compared to Barbari, despite higher mineral content in Sangak. Dephytinisation of both breads increased iron uptake in the Caco‐2 cells (0.65 vs. 1.64 in Sangak and 0.77 vs. 1.97 ng mg?1 protein in Barbari). Zinc uptake increased from 0.98 to 2.8 in Sangak and from 1.4 to 2.9 ng mg?1 protein in Barbari. Thus, dephytinisation substantially improves iron and zinc bioaccessibility.  相似文献   

15.
Arabinoxylans (AXs) from wheat malts potentially affect beer quality and production. β‐ d ‐Xylosidase is a key enzyme that degrades the main chains of AXs to produce xylose. This study performed a partial characterization of β‐ d ‐xylosidase from wheat malts. The optimal temperature was 70 °C and the enzyme exhibited excellent thermostability, that is, residual activities were 92.6% at 60 °C for 1 h. The enzyme was stable over a pH range of 3.0–6.0 and showed optimum activity at pH 3.5 and 4.5. Kinetic parameters Km and Vmax of wheat malt β‐ d ‐xylosidase against p‐nitrophenyl‐xyloside were 1.74 mmol L−1 and 0.76 m m min−1, respectively. The enzyme activity was severely inhibited by Cu2+, moderately inhibited by Mn2+, Mg2+, Al3+, Ca2+, Ba2+ and Na+ and mildly inhibited by Fe3+ and Fe2+. The partial enzymatic characterization achieved in this study can be used as a theoretical basis for purifying β‐ d ‐xylosidase from wheat malts. Copyright © 2015 The Institute of Brewing & Distilling  相似文献   

16.
Changes in the phytic acid, inorganic phosphorus and ATP contents, and in the activity of phytase and α-amylase in rice (Oryza sativa L) grains were determined during 18 days of germination in a dark room. The effect of phytic acid on α-amylase activity was studied in vitro. Rice grains immersed in sterilised deionised water at 14°C germinated on the fifth day. Phytase activity, detected in the ripening rice grains, increased linearly until the eighth day and reached a maximum on the tenth day. There was a marked decrease in phytate and an increase in inorganic phosphorus accompanying germination. There was a good inverse correlation between the levels of both phytase activity and inorganic phosphorus, and phytate breakdown. α-Amylase activity was detected on the fourth day and increased markedly from the 12th to the 16th day of germination. ATP level increased from the second to the fourth day and slightly decreased from the fourth to the eighth day; it increased rapidly again from the eighth to the 18th day of germination. α-Amylase activity was influenced by both pH and phytic acid concentration in the assay system. At 75 mM phytic acid, α-amylase activity was lowered by 23%, 93% and 52% at pH 4–0, 5–0 and 6–0 respectively. When the enzyme, phytate and Ca2+ were incubated together at pH 5–0, the inhibition of α-amylase by phytic acid was markedly decreased by addition of Ca2+. The chemical affinity of Ca2+ for phytic acid was higher in the reaction at pH 5–0 than in those at pH 4–0 and pH 6–0, and over 98% of Ca2+ in the reaction system was precipitated as Ca-phytate.  相似文献   

17.
The objectives of the current study were to determine S‐methyl‐L‐methionine (SMM) from various Brassicaceae family vegetables by using validated analytical method and to characterize the intestinal transport mechanism of SMM by the Caco‐2 cells. The SMM is well known to provide therapeutic activity in peptic ulcers. The amount of SMM from various Brassicaceae family vegetables ranged from 89.08 ± 1.68 μg/g to 535.98 ± 4.85 μg/g of dry weight by using validated ultra‐performance liquid chromatography‐electrospray ionization‐mass spectrometry method. For elucidating intestinal transport mechanism, the cells were incubated with or without transport inhibitors, energy source, or a metabolic inhibitor. Phloridzin and verapamil as inhibitors of sodium glucose transport protein (SGLT1) and P‐glycoprotein, respectively, were not responsible for cellular uptake of SMM. Glucose and sodium azide were not affected by the cellular accumulation of SMM. The efflux ratio of SMM was 0.26, implying that it is not effluxed through Caco‐2 cells. The apparent coefficient permeability (P app) of SMM was 4.69 × 10?5 cm/s, indicating that it will show good oral absorption in in vivo .  相似文献   

18.
In ruminants, more than 50% of overall gastrointestinal Ca absorption can occur preintestinally, and the anions of orally applied Ca salts are thought to play an important role in stimulating ruminal Ca absorption. This assumption is based mainly on ion-exchange studies that have used gluconate as the control anion, which may bind Ca2+ ions and interfere with treatment effects. In the present study, we investigated the distinct effects of different anions on Ca absorption across the sheep rumen and on the concentration of free Ca2+ ions ([Ca2+]ion). We showed that gluconate, sulfate, and short-chain fatty acids (SCFA) remarkably reduced [Ca2+]ion in buffer solutions. Nevertheless, increasing the Cl or SCFA concentration by 60 mM stimulated net ruminal Ca absorption 5- to 7-fold, but these effects could be antagonized by gluconate. Therefore, ion-exchange experiments must be (re)evaluated very carefully, because changes in [Ca2+]ion in the presence of gluconate, sulfate, or SCFA not only might entail an underestimation of Ca flux rates, but also might have effects on other cellular pathways that are Ca2+ dependent. Concerning the optimal Ca supply for dairy cows, the present study suggests that CaCl2 formulations and Ca salts of the SCFA stimulate Ca absorption across the rumen wall and are beneficial in preventing or correcting a Ca deficiency.  相似文献   

19.
Co‐flocculation between cells of S. cerevisiae NCYC 234 and NCYC 1109, both of which were non‐flocculent when cultivated in YM medium for 20 h, was investigated by chemical modification. Ca2+ promoted co‐flocculation. Protein‐denaturants and several carbohydrates caused reversible inhibition of the co‐flocculation in the presence of Ca2+. The effect of treatment with proteolytic enzymes and chemical modification of cell surface protein and carbohydrate components suggest strongly that co‐flocculation between cells of NCYC 234 and cells of NCYC 1109 results from interaction between surface protein component of cells of NCYC 1109 and surface carbohydrate component of cells of NCYC 234.  相似文献   

20.
A rapid and simple approach for visible determination of mercury ions (Hg2+) in aqueous solutions was developed based on surface plasmon resonance phenomenon using L‐arginine‐functionalized gold nanorods (AuNRs). At pH greater than 9, the deprotonated amine group of L‐arginine on the AuNRs bound with Hg2+ leading to the side‐by‐side assembly of AuNRs, which was verified by transmission electron microscopy images. Thus, when Hg2+ was present in the test solution, a blue shift of the typical longitudinal plasmon band of the AuNRs was observed in the ultra violet‐visible‐near infrared (UV‐Vis‐NIR) spectra, along with a change in the color of the solution, which occurred within 5 min. After carefully optimizing the potential factors affecting the performance, the L‐arginine/AuNRs sensing system was found to be highly sensitive to Hg2+, with the limit of detection of 5 nM (S/N = 3); it is also very selective and free of interference from 10 other metal ions (Ba2+, Ca2+, Cd2+, Co2+, Cs+, Cu2+, K+, Li+, Ni2+, Pb2+). The result suggests that the L‐arginine‐functionalized AuNRs can potentially serve as a rapid, sensitive, and easy‐to‐use colorimetric biosensor useful for determining Hg2+ in food and environmental samples.  相似文献   

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