体外模拟3 种消化液对铁皮石斛多糖的消化作用

本实验采用人体胃肠道模拟系统,体外模拟3 种消化液对铁皮石斛多糖的消化行为。通过高效凝胶液相色谱、铁氰化钾法和高效离子交换色谱分别测定消化后多糖分子质量变化、还原糖含量和单糖组成的情况。结果表明：唾液不能改变铁皮石斛多糖的分子质量;模拟胃液和模拟胃肠液可以降低多糖的分子质量,同时提高多糖溶液中的还原糖含量,但没有检测到单糖。结论：在体外模拟胃肠道的消化体系中,铁皮石斛多糖的分子质量减小,糖苷键发生断裂,但没有游离单糖的释放。     

体外模拟消化液对金钗石斛的作用

采用体外模拟消化液,分别对金钗石斛超微/普通粉、水/醇提物、粗石斛碱和多糖进行实验,探究实 验组模拟消化液中化学成分的变化。实验测定石斛碱、多糖溶出率;运用高效液相色谱、3,5-二硝基水杨酸法 和阴离子色谱对多糖消化产物的分子质量、还原糖质量浓度和单糖组成进行分析;采用液相色谱-质谱联用仪对 多酚类化合物成分进行分析。结果表明：消化组中石斛碱溶出率不断增大,超微粉肠道消化液组溶出率最高达 （21.46±0.52）%,粗石斛碱消化液最低为（8.74±0.50）%;水提物组多糖溶出率高,在口腔和胃消化液中,超 微粉多糖溶出率比普通粉高;肠道消化液中的情况与之相反;多糖在消化过程中分子质量降低,还原糖质量浓度上 升,无单糖检出;多酚类物质的质量浓度普遍在模拟胃液中升高,在模拟肠液中降低。     

GC‐MS‐Based Metabolite Profiling of Cosmos caudatus Leaves Possessing Alpha‐Glucosidase Inhibitory Activity

Cosmos caudatus, which is known as “Ulam Raja,” is an herbal plant used in Malaysia to enhance vitality. This study focused on the evaluation of the α‐glucosidase inhibitory activity of different ethanolic extracts of C. caudatus. Six series of samples extracted with water, 20%, 40%, 60%, 80%, and 100% ethanol (EtOH) were employed. Gas chromatography‐mass spectrometry (GC‐MS) and orthogonal partial least‐squares (OPLS) analysis was used to correlate bioactivity of different extracts to different metabolite profiles of C. caudatus. The obtained OPLS scores indicated a distinct and remarkable separation into 6 clusters, which were indicative of the 6 different ethanol concentrations. GC‐MS can be integrated with multivariate data analysis to identify compounds that inhibit α‐glucosidase activity. In addition, catechin, α‐linolenic acid, α‐D‐glucopyranoside, and vitamin E compounds were identified and indicate the potential α‐glucosidase inhibitory activity of this herb.     

Effect of In Vitro Digestion on Free α‐Dicarbonyl Compounds in Balsamic Vinegars

We investigated the influence of an in vitro simulated digestion process on the content of the free α‐dicarbonyl compounds most frequently found in food. A Glyoxal (GO), methylglyoxal (MGO), and diacetyl (DA) aqueous standard mixture and 2 brands of balsamic vinegar were analyzed before and after exposure to digestive enzymes. A strong matrix effect required adoption of validated RP‐HPLC‐DAD standard addition methods. The results showed that the digestive enzymes markedly alter the concentrations of the exogenous free α‐dicarbonyl compounds ingested with food; the extent of such changes varied with the α‐dicarbonyl compound itself and the diet components, which determined important but different food matrix effects also during digestion. The data also indicate that digestion can reduce the bioavailability of the toxic α‐dicarbonyl compounds ingested with food. However, no firm conclusions can be drawn about a putative positive influence of digestion on the toxic potential of dietary α‐dicarbonyl compounds, because their reaction in the presence of digestive enzymes likely gives rise to advanced glycation end products, which are involved in the development of chronic diseases.     

Chemical Constituents of Gold‐red Apple and Their α‐Glucosidase Inhibitory Activities

Ten compounds were isolated and purified from the peels of gold‐red apple (Malus domestica) for the 1st time. The identified compounds are 3β, 20β‐dihydroxyursan‐28‐oic acid (1), 2α‐hydroxyoleanolic acid (2), euscaphic acid (3), 3‐O‐p‐coumaroyl tormentic acid (4), ursolic acid (5), 2α‐hydroxyursolic acid (6), oleanolic acid (7), betulinic acid (8), linolic acid (9), and α‐linolenic acid (10). Their structures were determined by interpreting their nuclear magnetic resonance and mass spectrometry (MS) spectra, and by comparison with literature data. Compound 1 is new, and compound 2 is herein reported for the 1st time for the genus Malus. α‐Glucosidase inhibition assay revealed 6 of the triterpenoid isolates as remarkable α‐glucosidase inhibitors, with betulinic acid showing the strongest inhibition (IC₅₀ = 15.19 μM). Ultra‐performance liquid chromatography‐electrospray ionization MS analysis of the fruit peels, pomace, flesh, and juice revealed that the peels and pomace contained high levels of triterpenes, suggesting that wastes from the fruit juice industry could serve as rich sources of bioactive triterpenes.     

Electrospray MS‐based characterization of β‐carbolines – mutagenic constituents of thermally processed meat

The β‐carbolines 1‐methyl‐9H‐pyrido [3,4‐b]indole and 9H‐pyrido[3,4b]indole have been implicated as having causative roles in a number of human diseases, such as Parkinson's disease and cancer. As they can be formed during the heating of protein‐rich food, a number of analytical methodologies have been proposed for their detection and quantification in foodstuff. For this purpose, LC‐MS and LC‐MS/MS have emerged as the most specific analytical methods, and the quantification is based on the occurrence of unusual ions, such as [M+H‐(H^?)]⁺ and [M+H‐2H]⁺. In this study, we have investigated the formation of these ions by accurate‐mass electrospray MS/MS and demonstrated that these ions are formed from gas‐phase ion‐molecule reactions between water vapor present in the collision cell and the protonated molecule of 1‐methyl‐9H‐pyrido [3,4‐b]indole and 9H‐pyrido[3,4b]indole. Although this reaction has been previously described for heterocyclic amine ions, it has been overlooked in the most of recent LC‐MS and LC‐MS/MS studies, and no complete data of the fragmentation are reported. Our results demonstrate that additional attention should be given with respect to eliminating water vapor residues in the mass spectrometer when analysis of β‐carbolines is performed, as this residue may affect the reliability in the results of quantification.     

A preliminary study of monosaccharide composition and α‐glucosidase inhibitory effect of polysaccharides from pumpkin (Cucurbita moschata) fruit

In this work, crude polysaccharide extracts were extracted from pumpkin (Cucurbita moschata) fruit by hot water. After removal of proteins and purification, polysaccharides of pumpkin fruit (PP1‐1) were subjected to structural identification. Gas chromatography analysis indicated that PP1‐1 comprised of galactose (86.4%), and glucose (13.6%). The molecular weight of PP1‐1 was measured to be 0.87 × 10⁴ Da by gel permeation chromatography. The inhibitory kinetic evaluation showed that it was non‐competitive inhibition of PP1‐1 on the α‐glucosidase‐catalysed hydrolysis of PNPG. The Michaelis–Menten constant (K_m) was 0.106 m , and the inhibitory constants (K_i), 0.435 mg.     

Enzymatic hydrolysis combined with high‐pressure homogenisation for the preparation of polysaccharide‐based nanoparticles from the by‐product of Flammulina velutipes

In this study, protease, α‐amylase and 5‰ β‐glucanase enzymolysis in combination with high‐pressure homogenisation were used for the preparation of polysaccharide‐based nanoparticles from Flammulina velutipes stipes, respectively, named FNP‐1, FNP‐2 and FNP‐3, and the nanoparticles were subsequently characterised. The FNP size distribution ranged from 50 nm to 300 nm, among which FNP‐2 and FNP‐3 were smaller than FNP‐1, based on the SEM images. GC‐MS results showed that these particles were mainly composed of glucose and glucosamine. The FNP dispersions at 1 wt% behaved as non‐Newtonian, shear‐thinning fluids, and the FNP‐3 dispersion presented superior viscoelasticity. With an increasing degree of enzymolysis, the thermal stability of the FNPs decreased. In addition, these particles presented various cation‐exchange properties. Therefore, the Flammulina velutipes polysaccharide‐based nanoparticles obtained from this study can be potentially used as a promising functional food ingredient in the food industry.     

Effect of in vitro‐simulated gastrointestinal digestion on the stability and antioxidant activity of blueberry polyphenols and their cellular antioxidant activity towards HepG2 cells

In this study we investigated the influence of in vitro‐simulated gastrointestinal digestion on the bioaccessibility and antioxidant activity of polyphenols from blueberries (Vaccinium spp.). Total phenolic, anthocyanin, and flavonoid content was determined, and extract and digesta compositions were analysed by HPLC‐ESI‐MS/MS. The phenolic compounds were relatively stable under a gastric environment, whereas polyphenols and anthocyanins were unstable under an intestinal environment. The bioaccessibility of polyphenol, anthocyanin, and flavonoid was greatly decreased after the intestinal digestion, and the recoveries were only 13.93%, 1.95%, and 15.68% (the IN sample), respectively. Polyphenolic profile alteration occurred during in vitro‐simulated gastrointestinal digestion. Changes of phenolic compound antioxidant activity during digestion correlated with polyphenol, flavonoid, and caffeic acid concentrations. Digested extract cellular antioxidant activity was lower than non‐digested extract activity (P < 0.05). Polyphenol dose–response correlations with cellular antioxidant activity were observed. These results indicated that in vitro‐simulated gastrointestinal digestion significantly impact polyphenols and their antioxidant activity.     

Purification and physicochemical characterization of ovine β‐lactoglobulin and α‐lactalbumin

Ovine whey proteins were fractionated and studied by using different analytical techniques. Anion‐exchange chromatography and reversed‐phase high‐performance liquid chromatography (HPLC) showed the presence of two fractions of β‐lactoglobulin but only one of α‐lactalbumin. Gel permeation and sodium dodecyl sulfate (SDS)‐polyacrylamide gel electrophoresis allowed the calculation of the apparent molecular mass of each component, while HPLC coupled to electrospray ionisation‐mass spectrometry (ESI‐MS) technique, giving the exact molecular masses, demonstrated the presence of two variants A and B of ovine β‐lactoglobulin. Amino acid compositions of the two variants of β‐lactoglobulin differed only in their His and Tyr contents. Circular dichroism spectroscopy profiles showed pH conformation changes of each component. The thermograms of the different whey protein components showed a higher heat resistance of β‐lactoglobulin A compared to β‐lactoglobulin B at pH 2, and indicated high instability of ovine α‐lactalbumin at this pH.     

Physicochemical characterisation and α‐amylase inhibitory activity of tea polysaccharides under simulated salivary,gastric and intestinal conditions

Tea polysaccharides (TPS) are one of the main components of tea with various bioactivities. Digestion could result in the changes of the physicochemical properties and bioactivities of polysaccharides. The objective of this study was to determine the changes in physicochemical properties and α‐amylase inhibitory activities of TPS after simulated digestion. The structure of TPS determined by UV, IR and SEM showed that no obvious changes after salivary digestion, but significant changes after gastrointestinal digestion were observed. After intestinal digestion, the contents of reducing sugar were increased from 0.650 ± 0.011 to 1.594 ± 0.078 mm , and the centre molecular weight was decreased from 540.57 to 375.35 KDa. The molar ratio of monosaccharide was changed after digestion. And α‐amylase inhibition rate was significantly increased (P < 0.05). The results could provide information on the digestibility of TPSin vitro and contribute to the development of related functional foods or medicines.     

Influence of Lipid Content in a Corn Oil Preparation on the Bioaccessibility of β‐Carotene: A Comparison of Low‐Fat and High‐Fat Samples

Some individuals with fat maldigestion have compromised digestive systems, which causes the incomplete hydrolyzation of ingested lipids within the gastrointestinal tract (GIT). We studied the influence of high‐fat (20%) and low‐fat (4%) contents on the bioaccessibility of a highly hydrophobic nutraceutical (β‐carotene) through a simulated GIT model consisting of mouth, stomach, and small intestine phases. The low‐fat and high‐fat values were chosen to simulate low‐fat and high‐fat diets. The triglycerides in the low‐fat system were fully digested, whereas those in the high‐fat system were only partially digested, thereby mimicking the digestive systems of individuals who exhibit fat maldigestion. The carotenoids were initially solubilized within oil‐in‐water nanoemulsions prepared using a nonionic surfactant (Tween 20) as emulsifier and a long‐chain triglyceride (corn oil) as the oil phase. After digestion, the total β‐carotene concentration in the filtered micelle phase was much greater for the high‐fat group (0.072 μg/mL) than for the low‐fat group (0.032 μg/mL). Conversely, the β‐carotene bioaccessibility of the high‐fat group (39%) was much lower than that of the low‐fat group (84%), which was attributed to a fraction of the carotenoids remaining in the nondigested lipid phase of the high‐fat group. These results highlight the importance of delivering hydrophobic nutraceuticals in a form where the fat phase is fully digested.     

Comparison of structural,antioxidant and immuno‐stimulating activities of polysaccharides from Tremella fuciformis in two different regions of China

In this study, we compared the chemical structure, antioxidant and immuno‐stimulating activities of two polysaccharides, TPS–FJ and TPS–SC, extracted from Tremella fuciformis from two of the major cultivation regions of China. TPS–FJ and TPS–SC contained protein and high uronic acid. Average molecular weights of TPS–FJ and TPS–SC were 45 461 and 25 981 kDa, respectively. A triple‐helix conformation was exhibited by TPS–FJ but not by TPS–SC. TPS–FJ and TPS–SC contained same monosaccharides but in different ratios. Both the polysaccharides displayed characteristic polysaccharide bands in their Fourier transform infrared and NMR spectra, shown high water and oil holding capacity, and retained antioxidant potential after simulated gastrointestinal digestion in vitro. TPS–FJ significantly (P < 0.05) stimulated the nitric oxide, tumour necrosis factor‐α and interleukin‐6 production of RAW 264.7 cells with higher reactions than those of TPS–SC in vitro. Thus, Tremella fuciformis polysaccharide can be used as a potential antioxidant and immunomodulatory ingredient in food industry.     

Amylose and β‐Glucan Content of New Waxy Barleys

Starch was isolated from four new waxy barleys and compared with normal and high‐amylose barley starch. The waxy barley samples were selected lines from crosses of Swedish hulled and naked barley cultivars with the cultivar Azhul as donor of the waxy gene. The starches from the waxy barley samples were found to contain 0.7–2.6% amylose when determined iodimetrically by amperometric titration and 0.0–0.9% when determined by size exclusion chromatography after debranching. However, Sepharose CL‐2B elution profiles of the starches detected by iodine staining showed that all four waxy samples were free from detectable amounts of amylose. The amylopectin starches were found to contain a small polysaccharide fraction with molecular size smaller than amylopectin, with an iodine staining λ_max range of 550–600 nm. The water extractable and acid extractable β‐glucan contents in the waxy barley cultivars were generally found to be higher than those in normal barley.     

Release and characterization of single side chains of white cabbage pectin and their complement‐fixing activity

A mixture of single side chains from white cabbage pectin were obtained by anion exchange chromatography after applying mild chemical conditions promoting β‐elimination. These pectin fragments were characterized by their molecular weight distribution, sugar composition, ¹³C‐NMR, and MALDI‐TOF‐MS analysis. These analyses revealed that the large oligosaccharides released by β‐eliminative treatment were composed of α‐1,5 linked arabinosyl residues with 2‐ and 3‐linked α‐arabinosyl side chains, and, or β‐1,4 linked galactosyl side chains. Fractions were tested for complement‐fixing activity in order to determine their interaction with the complement system. These results strongly indicated that there was a minimal unit size responsible for the complement‐fixing activity. Neutral pectin fragments (?8 kDa) obtained from β‐elimination were inactive in the complement system, although they contained a sugar composition previously shown to be highly active. Larger pectin fragments (?17 kDa) retained some activity, but much lower than polymers containing rhamnogalacturonan type 1 (RGI) structures isolated from the same source. This implied that structural elements containing multiple side chains is necessary for efficient complement‐fixing activity.     

Isolation,structural features and rheological properties of water‐extractable β‐glucans from different Greek barley cultivars

β‐Glucans were isolated from six Greek barley cultivars (Persefoni, Kos, Thessaloniki, Athinaida, Dimitra and Triptolemos) by water extraction at 47 °C, enzymatic removal of starch and protein and subsequent precipitation of the water‐soluble β‐glucans with 37% (w/v) ammonium sulfate saturation. The purity of barley β‐glucans was high (>93% dry basis) with some small contamination by protein (<3.84%). The molecular size of the β‐glucan isolates was determined by high‐performance size‐exclusion chromatography (HPSEC); the weight‐average molecular weights and the intrinsic viscosities ranged between 0.45 × 10⁶ and 1.32 × 10⁶ and 2.77 and 4.11 dl g^?1, respectively. Structural features of barley β‐glucans were revealed by ¹³C NMR spectroscopy and high‐performance anion‐exchange chromatography (HPAEC) of the oligomers released by the hydrolytic action of lichenase. Lichenase degradation showed that β‐glucans from all barley cultivars consisted of blocks of cellotriosyl and cellotetraosyl units, accounting for 90.6–92.3% of the total oligomers released, with a molar proportion of these units between 2.31 and 2.77. Rheological measurements of aqueous solutions/dispersions of β‐glucans showed the behaviour of non‐interacting polysaccharides and a transition from the typical viscoelastic response to gel‐like properties after a time period that depended on the molecular size of the polysaccharide. The lowest molecular size β‐glucans from the Triptolemos cultivar showed shorter gelation times than their higher molecular weight counterparts. The effect of sugar incorporation (glucose, fructose, sucrose, xylose and ribose), at a concentration of 30% (w/v), to the β‐glucans gels (6% w/v) on compression parameters seemed to be related to the type of sugar used; the pentose sugars substantially reduced gel firming. Copyright © 2004 Society of Chemical Industry     

Enzymatic Properties of β‐1,3‐Glucanase from Streptomyces sp Mo.

Streptomyces sp Mo endo‐β‐1,3‐glucanase was found to have hydrolyzing activity toward curdlan and released laminarioligosaccharides selectively. The molecular weight was estimated to be 36000 Da and its N‐terminal amino acid sequence was VTPPDISVTN. The optimal pH was 6 and the enzyme was found to be stable from pH 5 to 8. The optimal temperature was 60 °C and the activity was stable below 50 °C. The enzyme hydrolyzed selectively curdlan containing only β‐1,3 linkages. The enzyme had 89% relative activity toward Laminaria digitata laminarin, which contains a small amount of β‐1,6 linkages compared with curdlan, while Eisenia bicyclis laminarin with a higher amount of β‐1,6‐linkages, was not hydrolyzed. Mo enzyme adsorbed completely on curdlan powder. The enzymatic hydrolysis of curdlan powder resulted in the accumulation of laminaribiose (yield 81.7%). Trisaccharide was inevitably released from the hydrolysis of laminarioligosaccharides with 5 to 7 degrees of polymerization (DP). Although the enzyme cleaved off disaccharide (DP 2) from tetrasaccharide (DP 4), the reaction rate was lower than those of DP 5 to 7. The results indicated that the active site of Mo endo‐β‐1,3‐glucanase can efficiently recognize glucosyl residue chain of greater than DP 5 and hydrolyzes the β‐1,3 linkage between the 3rd and 4th glucosyl residue.     

Bioaccessibility,cellular uptake and transepithelial transport of α‐tocopherol and retinol from a range of supplemented foodstuffs assessed using the caco‐2 cell model

The bioaccessibility, or amount of a nutrient available for gastrointestinal absorption, can be determined using an in vitro digestion model, the addition of the resultant digestate to a caco‐2 transwell model system yields an approximation of nutrient bioavailability. The objective of the present study was to compare the bioaccessibility and bioavailability of α‐tocopherol and retinol from a range of digested foodstuffs. Minced pork, beef and turkey and apple sauce, bread and mayonnaise were supplemented with α‐tocopherol‐acetate and retinol‐acetate prior to being subjected to an in vitro digestion procedure. The aqueous fraction of each of the digested foodstuffs was then added to a caco‐2 transwell model and the transepithelial transport was determined. The findings of the present study indicate that α‐tocopherol and retinol are more bioaccessible from supplemented meat products than from supplemented apple sauce, bread and mayonnaise. It was found that turkey meat facilitated the highest bioaccessibility and subsequent cellular uptake and transport of retinol. The cellular uptake and secretion of α‐tocopherol was similar for all samples.     

Screening of β‐Glucosidase and β‐Xylosidase Activities in Four Non‐Saccharomyces Yeast Isolates

The finding of new isolates of non‐Saccharomyces yeasts, showing beneficial enzymes (such as β‐glucosidase and β‐xylosidase), can contribute to the production of quality wines. In a selection and characterization program, we have studied 114 isolates of non‐Saccharomyces yeasts. Four isolates were selected because of their both high β‐glucosidase and β‐xylosidase activities. The ribosomal D1/D2 regions were sequenced to identify them as Pichia membranifaciens Pm7, Hanseniaspora vineae Hv3, H. uvarum Hu8, and Wickerhamomyces anomalus Wa1. The induction process was optimized to be carried on YNB‐medium supplemented with 4% xylan, inoculated with 106 cfu/mL and incubated 48 h at 28 °C without agitation. Most of the strains had a pH optimum of 5.0 to 6.0 for both the β‐glucosidase and β‐xylosidase activities. The effect of sugars was different for each isolate and activity. Each isolate showed a characteristic set of inhibition, enhancement or null effect for β‐glucosidase and β‐xylosidase. The volatile compounds liberated from wine incubated with each of the 4 yeasts were also studied, showing an overall terpene increase (1.1 to 1.3‐folds) when wines were treated with non‐Saccharomyces isolates. In detail, terpineol, 4‐vinyl‐phenol and 2‐methoxy‐4‐vinylphenol increased after the addition of Hanseniaspora isolates. Wines treated with Hanseniaspora, Wickerhamomyces, or Pichia produced more 2‐phenyl ethanol than those inoculated with other yeasts.