Enzymatic production of galactooligosaccharides by beta-galactosidase from Bifidobacterium longum BCRC 15708

The production of galactooligosaccharides (GOSs) by transgalactosylation using beta-galactosidase from Bifidobacterium longum BCRC 15708 was studied. Other than lactose, galactose, and glucose, two types of GOSs, tri- and tetrasaccharides, were formed after beta-galactosidase action on 40% lactose. Trisaccharides were the major type of GOS formed. Generally, an increase of the initial lactose concentration in the reaction mixture resulted in a higher GOS production. A maximum yield of 32.5% (w/w) GOSs could be achieved from 40% lactose solution at 45 degrees C, pH 6.8, when the lactose conversion was 59.4%. The corresponding productivity of GOSs was 13.0 g/(L.h). Transgalactosylation activity of beta-galactosidase from a test organism showed a relatively lower sensitivity toward glucose and galactose than that from other organisms. The addition of 5% or 10% glucose or galactose to the reaction mixture did not significantly (p>0.05) reduce the transgalactosylation reaction of beta-galactosidase.     

Oxymyoglobin oxidation and membranal lipid peroxidation initiated by iron redox cycle.

Oxymyoglobin is the main pigment in muscle tissues, responsible for the bright red color of fresh meat. Oxidation of the heme iron from the ferrous to the ferric metmyoglobin produces the brownish color that consumers find undesirable in fresh meat. The aim of this study was to elucidate the mechanism of oxymyoglobin oxidation in muscle tissues by using a model system containing oxymyoglobin and muscle membranes oxidized by an iron redox cycle. Oxidation of oxymyoglobin was determined from the decrease in absorption of the solution measured by a spectrophotometer at 582 nm. Lipid peroxidation was determined by accumulation of TBARS and conjugated dienes. The higher rates of oxidation of oxymyoglobin (20 microM) and lipid oxidation were achieved by using ferric iron and ascorbic acid at concentrations of 50 and 200 microM, respectively. Increasing the concentration of ascorbic acid to 2000 microM switched its effect to antioxidative. Increasing the concentration of oxymyoglobin from 20 to 80 microM inhibited lipid peroxidation by >90% and partially prevented oxymyoglobin oxidation.     

Characterization of exopolysaccharides produced by Bifidobacterium longum NB667 and its cholate-resistant derivative strain IPLA B667dCo

Bifidobacteria are natural members of the human intestinal microbiota and some strains are being used as probiotics. Adaptation to bile can allow them to increase survival in gastrointestinal conditions, thus improving their viability. Bifidobacterium longum NB667 and the cholate-resistant strain B. longum IPLA B667dCo produced exopolysaccharides (EPS) that were partially characterized. Analysis by size exclusion chromatography-multiangle laser light scattering indicated that the EPS crude fractions of both strains contained two polymer peaks of different molar mass. On the basis of chromatographic techniques both peaks appeared to be heteropolysaccharides. The smaller peak was mainly composed of glucose, galactose and rhamnose whose molar ratios and linkage types showed slight variations between the EPS fractions of both strains. The bigger peak consisted of glucose and galactose; the monosaccharide composition was identical in the EPS fractions of the two microorganisms, but their infrared spectra presented some differences regarding compounds other than carbohydrates that seem to be associated to the polymer. Differences in the composition of EPS fractions did not affect the capability of crude EPS from B. longum to be fermented by the human intestinal microbiota in fecal batch cultures.     

Influence of Bifidobacterium longum CECT 7347 and gliadin peptides on intestinal epithelial cell proteome

Celiac disease is an enteropathy caused by an abnormal immune response to cereal gluten proteins (gliadin). To unravel the possible role of the interactions between gliadin peptides and specific intestinal bacteria, the response of intestinal epithelial (Caco-2) cells to gliadin subjected to gastrointestinal digestion in the presence or absence of Bifidobacterium longum CECT 7347 has been studied. Changes in the proteome of Caco-2 cells were determined by 2DE and MALDI-TOF. Gliadins digested without B. longum altered the expression of a higher number of proteins than in the presence of the bacterium (21 versus 9), and these proteins were involved in disorganization of cell cytoskeleton, inflammation, and apoptosis. Gliadins digested in the presence of the bacterium influenced the production of proteins involved in calcium homeostasis and cell survival and function. Therefore, B. longum CECT 7347 might ameliorate gliadin toxicity and modify the responses of intestinal epithelial cells to the gliadin challenge.     

Inhibition of lipid peroxidation and structure-activity-related studies of the dietary constituents anthocyanins,anthocyanidins, and catechins

The antioxidant activities of a series of commonly consumed and biogenetically related plant phenolics, namely, anthocyanidins, anthocyanins, and catechins, in a liposomal model system have been investigated. The antioxidant efficacies of the compounds were evaluated on their abilities to inhibit the fluorescence intensity decay of an extrinsic probe, 3-[p-(6-phenyl)-1,3,5-hexatrienyl]phenylpropionic acid, caused by free radicals generated during metal ion-induced peroxidation. Distinct structure-activity relationships were revealed for the antioxidant abilities of these structurally related compounds. Whereas antioxidant activity increased with an increasing number of hydroxyl substituents present on the B-ring for anthocyanidins, the converse was observed for catechins. However, substitution by methoxyl groups diminished the antioxidant activity of the anthocyanidins. Substitution at position 3 of ring C played a major role in determining the antioxidant activity of these classes of compounds. The anthocyanidins, which possess a hydroxyl group at position 3, demonstrated potent antioxidant activities. For the cyanidins, an increasing number of glycosyl units at position 3 resulted in decreased antioxidant activity. Similarly, the substitution of a galloyl group at position 3 of the flavonoid moiety resulted in significantly decreased antioxidant activity for the catechins. Among catechins, cis-trans isomerism, epimerization, and racemization did not play a role in overall antioxidant activity. The antioxidant activities of test compounds (at 40 microM concentrations) were compared to the commercial antioxidants tert-butylhydroquinone, butylated hydroxytoluene, butylated hydroxyanisole, and vitamin E (all at 10 microM concentrations).     

嗜酸乳杆菌真空喷雾干燥微胶囊化技术的研究

以阿拉伯胶、β-环糊精、可溶性淀粉、明胶、脱脂奶粉为壁材，根据正交试验确定微生态菌株嗜酸乳杆菌细胞微胶囊化的最佳真空喷雾干燥进风温度、壁材浓度和芯材浓度，并对其粒子直径进行分析。结果表明，β-环糊精与阿拉伯胶以9∶1配比为理想壁材，喷雾干燥进风温度170℃、壁材浓度20%和芯材浓度6%为最佳组合，活菌数达10⁸ cfu/g以上，微胶囊粒子直径平均为20 μm。     

Inhibition of diacylglycerol acyltransferase by alkamides isolated from the fruits of Piper longum and Piper nigrum

Pharmacological inhibition of acyl CoA:diacylglycerol acyltransferase (DGAT, EC 2.3.1.20) has emerged as a potential therapy for the treatment of obesity and type 2 diabetes. Bioassay-guided isolation of CHCl3 extracts of the fruits of Piper longum and Piper nigum (Piperaceae), using an in vitro DGAT inhibitory assay, lead to isolation of a new alkamide named (2E,4Z,8E)-N-[9-(3,4-methylenedioxyphenyl)-2,4,8-nonatrienoyl]piperidine (2), together with four known alkamides: retrofractamide C (1), pipernonaline (3), piperrolein B (4), and dehydropipernonaline (5). Compounds 2-5 inhibited DGAT with IC50 values of 29.8 (2), 37.2 (3), 20.1 (4), and 21.2 (5) microM, respectively, but the IC50 value for 1 was more than 900 microM. This finding indicates that compounds possessing piperidine groups (2-5) can be potential DGAT inhibitors.     

Enhanced survival of probiotic Lactobacillus acidophilus by encapsulation with nanostructured polyelectrolyte layers through layer-by-layer approach

The encapsulation of probiotic Lactobacillus acidophilus through layer-by-layer self-assembly of polyelectrolytes (PE) chitosan (CHI) and carboxymethyl cellulose (CMC) has been investigated to enhance its survival in adverse conditions encountered in the GI tract. The survival of encapsulated cells in simulated gastric (SGF) and intestinal fluids (SIF) is significant when compared to nonencapsulated cells. On sequential exposure to SGF and SIF for 120 min, almost complete death of free cells is observed. However, for cells coated with three nanolayers of PEs (CHI/CMC/CHI), about 33 log?% of the cells (6?log?cfu/500?mg) survived under the same conditions. The enhanced survival rate of encapsulated L. acidophilus can be attributed to the impermeability of polyelectrolyte nanolayers to large enzyme molecules like pepsin and pancreatin that cause proteolysis and to the stability of the polyelectrolyte nanolayers in gastric and intestinal pH. The PE coating also serves to reduce viability losses during freezing and freeze-drying. About 73 and 92 log?% of uncoated and coated cells survived after freeze-drying, and the losses occurring between freezing and freeze-drying were found to be lower for the coated cells.     

Improved bioavailability of dietary phenolic acids in whole grain barley and oat groat following fermentation with probiotic Lactobacillus acidophilus , Lactobacillus johnsonii , and Lactobacillus reuteri

The aim of this study was to improve the bioavailability of the dietary phenolic acids in flours from whole grain barley and oat groat following fermentation with lactic acid bacteria (LAB) exhibiting high feruloyl esterase activity (FAE). The highest increase of free phenolic acids was observed after fermentation with three probiotic strains, Lactobacillus johnsonii LA1, Lactobacillus reuteri SD2112, and Lactobacillus acidophilus LA-5, with maximum increases from 2.55 to 69.91 μg g(-1) DM and from 4.13 to 109.42 μg g(-1) DM in whole grain barley and oat groat, respectively. Interestingly, higher amounts of bound phenolic acids were detected after both water treatment and LAB fermentation in whole grain barley, indicating higher bioaccessibility, whereas some decrease was detected in oat groat. To conclude, cereal fermentation with specific probiotic strains can lead to significant increase of free phenolic acids, thereby improving their bioavailability.     

Detection of lipid peroxidation catalyzed by chelated iron and measurement of antioxidant activity in wine by a chemiluminescence analyzer.

We measured directly the reactive oxygen generated from a peroxide-free reaction system when a ferrous complex with nitrilotriacetic acid was oxidized to the ferric complex. Further, it was observed by a measurement of chemiluminescence that peroxidation of a lipid substrate added in the system is initiated by the Fe(3+)-type of reactive oxygen generated. Antioxidant activity can be estimated by contrasting the reaction rates of lipid peroxidation between the systems with and without a putative antioxidant sample. By this method, the antioxidant activity, expressed as catechin equivalent, of red wines for linoleic acid peroxidation was shown to be higher than those of rosé and white wines (189-311, 84, and 37 microM for red, rosé, and white wines, respectively) because of a higher concentration of polyphenols such as flavanol and anthocyanin in red wines. The chemiluminescence measurement would be a promising method for evaluating the antioxidant potential because of its highly specific and sensitive detection of the hydroperoxide and for monitoring in situ peroxidation reaction.     

产细菌素的嗜酸乳杆菌WS发酵动力学模型的建立(简报)

为了嗜酸乳杆菌进一步工业化生产,需要确定嗜酸乳杆菌的发酵动力学模型,该文将嗜酸乳杆菌WS接种到MRS培养基进行发酵培养,根据其发酵过程特点,在Logistic方程和Luedeking-Pitet方程的基础上,建立了嗜酸乳杆菌WS发酵过程中菌体生长、基质消耗和产物形成的动力学模型,并用实际发酵实验值进行验证.结果表明,模型预测值和实验值吻合较好,说明该文建立的模型能较好地预测实际的发酵过程.     

Oxymyoglobin oxidation and membrane lipid peroxidation initiated by iron redox cycle: prevention of oxidation by enzymic and nonenzymic antioxidants.

The red color of muscle is principally due to the presence of oxymyoglobin. Oxidation of heme iron from the ferrous to the ferric state produces a brownish color, which consumers find undesirable. The aim of this study was to use enzymic and nonenzymic antioxidants to simulate in situ muscle antioxidation reactions in order to understand better the mechanism by which the iron redox cycle catalyzes membrane lipid peroxidation and oxymyoglobin oxidation. The inclusion of superoxide dismutase (SOD) in the model system decreased oxymyoglobin oxidation by 10% without affecting lipid peroxidation. Addition of catalase decreased oxymyoglobin oxidation by approximately 40% but not lipid peroxidation. Increasing the ceruloplasmin concentration inhibited lipid peroxidation but increased oxymyoglobin oxidation, which was inhibited by SOD and catalase. Conalbumin (50 microM), a specific iron chelator, inhibited peroxidation and oxymyoglobin oxidation by almost 50%. The addition of the antioxidant catechin (500 microM) decreased lipid peroxidation by 90% but oxymyoglobin oxidation by only 50%. Feeding turkeys with vitamin E at several levels significantly increased the alpha-tocopherol level of membranes, thus preventing oxymyoglobin and lipid oxidation. In conclusion, oxymyoglobin stability in the model system was affected by two pathways: (a) oxygen active species, such as O(2)*(-), H(2)O(2), HO*, and ferryl, generated during autoxidation of myoglobin and oxidation of ferrous ions and ascorbic acid; and (b) lipid radicals, such as ROO*, RO*, and hydroperoxides, generated during lipid peroxidation. Maximum inhibition could be achieved only by introducing inhibitors of both pathways into the system.     

Inhibition of protein and lipid oxidation in liposomes by berry phenolics

The antioxidant activity of berry phenolics (at concentrations of 1.4, 4.2, and 8.4 mug of purified extracts/mL of liposome sample) such as anthocyanins, ellagitannins, and proanthocyanidins from raspberry (Rubus idaeus), bilberry (Vaccinium myrtillus), lingonberry (Vaccinium vitis-idaea), and black currant (Ribes nigrum) was investigated in a lactalbumin-liposome system. The extent of protein oxidation was measured by determining the loss of tryptophan fluorescence and formation of protein carbonyl compounds and that of lipid oxidation by conjugated diene hydroperoxides and hexanal analyses. The antioxidant protection toward lipid oxidation was best provided by lingonberry and bilberry phenolics followed by black currant and raspberry phenolics. Bilberry and raspberry phenolics exhibited the best overall antioxidant activity toward protein oxidation. Proanthocyanidins, especially the dimeric and trimeric forms, in lingonberries were among the most active phenolic constituents toward both lipid and protein oxidation. In bilberries and black currants, anthocyanins contributed the most to the antioxidant effect by inhibiting the formation of both hexanal and protein carbonyls. In raspberries, ellagitannins were responsible for the antioxidant activity. While the antioxidant effect of berry proanthocyanidins and anthocyanins was dose-dependent, ellagitannins appeared to be equally active at all concentrations. In conclusion, berries are rich in monomeric and polymeric phenolic compounds providing protection toward both lipid and protein oxidation.     

Ameliorative influence of sesame lignans on lipid profile and lipid peroxidation in induced diabetic rats

Sesame lignans are working as antioxidants in various physiological functions. In the present study, the antioxidative effect of sesame lignans is examined in chemically induced diabetes mellitus (DM) in rats against lipid profile and lipid peroxidations. DM was induced in four groups of rats by injection of alloxan. The control groups (non-diabetic and diabetic) received a diet containing sunflower oil while the rest of the three experimental diabetic groups received a diet containing 0.25% alpha-tocopherol (D-Toc), 0.5% sesame lignan (D-SL), and 0.25% alpha-tocopherol+0.25% sesame lignan (D-Toc-SL) in sunflower oil for 4 weeks. Lipid profile and lipid peroxidations of plasma, erythrocyte membrane (EM), and liver tissues were measured. The total cholesterol, non-HDL cholesterol, plasma lipid peroxidation, and also LDL-peroxidation decreased, and HDL cholesterol increased significantly (P<0.05) in all the experimental groups as compared to the control diabetic sunflower oil group. The triacylglycerol (TAG) level in plasma decreased significantly in the D-SL and D-Toc-SL groups as compared to control diabetic group. Significant decrease in TAG level was observed in the D-SL group as compared to the D-Toc group. LDL peroxidation also decreased significantly in the D-Toc-SL group as compared to the D-Toc group. EM lipid peroxidation and liver lipid peroxidation decreased significantly in the D-Toc, D-SL, and D-Toc-SL groups as compared to the control diabetic group. Liver TAG level decreased more significantly in the D-SL and D-Toc-SL groups than in the control diabetic group. So, sesame lignans at 0.5% level and sesame lignan + alpha-tocopherol significantly ameliorate the alteration in lipid profile and the adverse free radical generative influence of DM induced by alloxan.     

Mechanism of inhibition of lipid peroxidation by gamma-terpinene,an unusual and potentially useful hydrocarbon antioxidant

gamma-Terpinene (TH), a monoterpene hydrocarbon present in essential oils, retards the peroxidation of linoleic acid (LH). The peroxidation of TH has been shown to yield p-cymene as the only organic product in a chain reaction in which the chain carrier is the hydroperoxyl radical, HOO(.). The peroxidation of LH is well-known to be a chain reaction in which the chains are carried by linoleylperoxyl radicals, LOO., and the products are linoleyl hydroperoxides. The retardation of LH peroxidation by TH has been found to be due to rapid chain termination via a very fast cross-reaction between HOO. and LOO. radicals. This antioxidant mechanism is completely different from the mechanism of antioxidant action of vitamin E. Since vitamin E becomes a prooxidant at high concentrations, the addition of essential oils containing TH to edible lipids may provide an alternative or supplementary strategy for obtaining large increases in their oxidative stability and shelf life, something that cannot be achieved by simply adding more and more vitamin E.     

Assessment of radical scavenging capacity and lipid peroxidation inhibiting capacity of antioxidant

The role of radical scavenging antioxidants against oxidative stress has received much attention, and the antioxidant capacity has been assessed by various methods. Among them, a method that measures the effect of antioxidant on decay of the probe is one of the most widely used methods. The present study was performed to compare the two methods to assess the antioxidant capacity, one to follow the decay of the probe and the other to measure lipid peroxidation products in human plasma. It was shown that the method following probe decay was suitable for assessment of radical scavenging capacity of antioxidant, but not for the capacity to inhibit lipid peroxidation in plasma. This is true whether a hydrophilic or lipophilic probe is used. Such different results arise from the fact that the efficacy of inhibition of lipid peroxidation by antioxidants depends on the fate of antioxidant-derived radical and interaction between antioxidants as well as the capacity of free radical scavenging. Thus, the capacity of antioxidants for inhibition of lipid peroxidation should be assessed from the effect on the extent of oxidation, not from the effect on probe decay.     

Synthesis of lipophilic clovamide derivatives and their antioxidative potential against lipid peroxidation

Some N-(hydroxycinnamoyl)-L-tyrosine and L-DOPA alkyl esters were synthesized and evaluated as a variation of the clovamide (N-caffeoyl-L-3,4-dihydroxyphenylalanine) structure, a known antioxidant found in red clover. The amides were prepared in good yields starting from methyl and dodecylesters of L-tyrosine and L-DOPA by reacting with the N-hydroxysuccinimidyl esters of ferulic, sinapic, and acetyl-protected caffeic acid, respectively. In the DPPH* (2,2-diphenyl-1-picrylhydrazyl) and superoxide radical quencher assays they showed radical scavenging activity equal to or higher than those of the standard antioxidants ascorbic acid and tocopherol. The antioxidative potentials of the clovamide derivatives against bulk lipid oxidation, as determined by the accelerated autoxidation of oils, were equal to or higher than those of the standard antioxidants; some of the compounds were able to protect an emulsion of linoleic acid/beta-carotene against oxidation. N-Caffeoyl L-tyrosine methyl ester and the N-cinnamoyl L-DOPA alkyl esters especially were potent antioxidants in bulk lipids and moderate protectants in emulsions.     

Inhibition of lipid oxidation in pork bundles processing by superheated steam frying

The effect of superheated steam treatment on the oxidative stability of lipids in packaged Zousoon (pork bundles) was investigated. The aroma quality of Zousoon samples was evaluated by sensory analysis and chromatographic analysis of volatiles. Results of this study indicated that oxidation of lipids occurred in pan-fried Zousoon after prolonged storage. Significant amounts of highly volatile compounds such as formaldehyde, acetaldehyde, acetone, and hexanal in Zousoon were identified by a modified method of cysteamine derivatization followed by gas chromatography-mass spectrometry (GC-MS) analysis. Superheated steam was found to be effective in suppressing lipid oxidation in canned Zousoon as compared with Zousoon fried by the conventional method in a frying pan. The superheated steam-fried samples had relatively low thiobarbituric acid reactive substance (TBARS) and peroxide (POV) values before and after storage, whereas samples prepared by pan frying had relatively high TBARS and POV values before and after storage. Superheated steam-fried Zousoon had superior lipid stability to that prepared by the conventional pan-frying method.     

Efficacy of metmyoglobin and hemin as a catalyst of lipid peroxidation determined by using a new testing system

A new quantitative approach to investigate the capability of iron heme complexes (HEM), metmyoglobin and hemin, to catalyze lipid peroxidation was elaborated. The oxidation of methyl linoleate in micellar solutions was used as a testing model. The key point was the determination of the rate of free radical generation, RIN, calculated from the rate of oxygen consumption. The HEM catalytic activity was characterized by two independent parameters: by reactivity and by its resistance to degradation. Both parameters were found to be pH-dependent. The reactivity was expressed as the effective rate constant for the reaction of HEM with lipid hydroperoxide. The resistance to degradation was characterized by the rate of the decrease in RIN with time and also by the regeneration coefficient, which shows how many active free radicals can be generated by one molecule of HEM. Both Hemin and metMB were found to be very effective catalysts even at nanomolar concentrations. The effective regeneration of active forms of HEM was observed. The catalytic activity of HEM was rapidly reduced with time. The kinetic scheme of the process under consideration was suggested, and this was applied for kinetic computer simulations.     

Relative inhibition of lipid peroxidation, cyclooxygenase enzymes, and human tumor cell proliferation by natural food colors

The most abundant water soluble natural food colors are betacyanins and anthocyanins. Similarly, lycopene, bixin, beta-carotene, and chlorophyll are water insoluble colors. Pure betanin, bixin, lycopene, chlorophyll, beta-carotene, and cyanidin-3-O-glucoside were isolated from Beta vulgaris, Bixa orellana,Lycopersicum esculentum, Spinacia oleracea, Daucus carrota, and Prunus cerasus, respectively. These natural pigments, alone and in combination, were evaluated for their relative potencies against cyclooxygenase enzymes and tumor cell growth inhibition by using MCF-7 (breast), HCT-116 (colon), AGS (stomach), CNS (central nervous system), and NCI-H460 (lung) tumor cell lines. Among the colors tested, betanin, cyanidin-3-O-glucoside, lycopene, and beta-carotene inhibited lipid peroxidation. However, all pigments tested gave COX-1 and COX-2 inhibition and showed a dose-dependent growth inhibition against breast, colon, stomach, central nervous system, and lung tumor cells, respectively. The mixtures of these pigments were also evaluated for their synergistic effects and chemical interactions at various concentrations. The mixture of anthocyanin and betanin negated their efficacy in the cell growth inhibitory assay and did not enhance the COX enzyme inhibitory activity. This is the first report of a comparative evaluation and the impact on biological activities of these pigments alone and in combination.