Brown seaweed as a food ingredient contributing to an adequate but not excessive amount of iodine in the European diet. A case study with bread.

BACKGROUND: The large amounts of iodine in brown seaweeds may contribute towards an adequate iodine intake, but also pose a food safety risk. In the current work we estimate the maximum amount of the cultivated brown seaweeds Saccharina latissima and Alaria esculenta (blanched and non-blanched) that can be added to white loaf-type wheat-bread so European consumers are protected against excessive chronic intakes. We use data for high-level consumers of bread with special ingredients added from the EFSA comprehensive European food consumption database to construct a conservative risk management model. We bake prototype seaweed bread and use the output from the model to assess exposure to iodine and trace metals. We also assess some bread quality parameters such as sensory characteristics, crumb firmness and specific volume. RESULTS: The maximum level of iodine in bread that would mitigate the European consumer from excess intakes was 857 µg I kg-1 bread. Assuming a typical 60% wheat flour per kilogram of bread, a maximum amount of 11.3 g of dried blanched A. esculenta (115 mg I kg-1 dry weight) could be incorporated per kilogram of wheat flour into the bread recipe whilst for non-blanched S. latissima, which contained 3500 mg I kg-1 dry weight, only a minuscule 350 mg of seaweed was the limit. For prototype breads, seaweed addition had no significant impact on the specific volume and crumb firmness. Levels of cadmium and arsenic in the bread were also of low toxicological concern. CONCLUSION: A maximum 1-2% brown seaweed of total flour weight could be added to bread to contribute towards European consumers' adequate iodine intake without overexposure. © 2024 The Author(s). Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

A Three-Day Intervention With Granola Containing Cereal Beta-Glucan Improves Glycemic Response and Changes the Gut Microbiota in Healthy Individuals: A Crossover Study.

Intake of soluble fibers including beta-glucan, is known to improve post-prandial glycemic response. The mechanisms have been attributed to the viscous gel forming in the stomach and small intestine, giving a longer absorption time. However, recent evidence suggests a link between intake of beta-glucan and improved glycemic regulation at subsequent meals through the gut microbiota. We investigated the short-term effect of granola with different amounts of cereal beta-glucan on glycemic response and gut microbiota. After a two-week run-in period (baseline), fourteen healthy, normal weight adults completed a dose-response dietary crossover study. Different amounts of cereal beta-glucan (low: 0.8 g, medium: 3.2 g and high: 6.6 g) were provided in granola and eaten with 200 ml low-fat milk as an evening meal for three consecutive days. Blood glucose and insulin were measured fasted and after an oral glucose tolerance test (OGTT) the following day, in addition to peptide YY (PYY) and glucagon-like peptide (GLP-2), fasting short chain fatty acids (SCFA) in blood, breath H2, and gut microbiota in feces. Only the intervention with medium amounts of beta-glucan decreased blood glucose and insulin during OGTT compared to baseline. Fasting PYY increased with both medium and high beta-glucan meal compared to the low beta-glucan meal. The microbiota and SCFAs changed after all three interventions compared to baseline, where acetate and butyrate increased, while propionate was unchanged. Highest positive effect size after intake of beta-glucan was found with Haemophilus, followed by Veillonella and Sutterella. Furthermore, we found several correlations between different bacterial taxa and markers of glycemic response. In summary, intake of granola containing 3.2 g cereal beta-glucan as an evening meal for three consecutive days reduced the glycemic response after an OGTT 0-180 min and changed gut microbiota composition. Since we cannot rule out that other fiber types have contributed to the effect, more studies are needed to further explore the effect of cereal beta-glucan on glycemic regulation. Clinical Trial Registration: [www.clinicaltrials.gov], identifier [NCT03293693].

Adjuvant Effect of Orally Applied Preparations Containing Non-Digestible Polysaccharides on Influenza Vaccination in Healthy Seniors: A Double-Blind, Randomised, Controlled Pilot Trial.

Senior individuals can suffer from immunosenescence and novel strategies to bolster the immune response could contribute to healthy ageing. In this double-blind, randomised, controlled pilot trial, we investigated the ability of non-digestible polysaccharide (NPS) preparations to enhance the immune response in a human vaccination model. In total, 239 subjects (aged 50-79 years) were randomised to consume one of five different NPS (yeast ß-glucan (YBG), shiitake ß-glucan (SBG), oat ß-glucan (OBG), arabinoxylan (AX), bacterial exopolysaccharide (EPS)) or control (CTRL) product daily for five weeks. After two weeks of intervention, subjects were vaccinated with seasonal influenza vaccine. The post-vaccination increases in haemagglutination inhibition antibody titres and seroprotection rate against the influenza strains were non-significantly enhanced in the NPS intervention groups compared to CTRL. Specifically, a trend towards a higher mean log2 fold increase was observed in the AX group (uncorrected p = 0.074) combined with a trend for an increased seroprotection rate, AX group (48.7%) compared to CTRL (25.6%) (uncorrected p = 0.057), for the influenza A H1N1 strain. Subjects consuming AX also had a reduced incidence of common colds compared to CTRL (1 vs. 8; p = 0.029 in Fisher exact test). No adverse effects of NPS consumption were reported. The findings of this pilot study warrant further research to study AX as an oral adjuvant to support vaccine efficacy.

Improved estimation of in vitro protein digestibility of different foods using size exclusion chromatography.

While the harmonized INFOGEST model provides a physiologically relevant platform for simulated digestion, it needs to be combined with adequate analytical methods to enable quantification and comparison of protein digestibility in different food matrices. We have shown that size exclusion chromatography (SEC) can be used to estimate the proportion of small peptides potentially available for uptake. Combined with determination of total dissolved protein, the % of small peptides per total protein was calculated as a physiologically relevant estimate of protein digestibility (DSEC). Values for DSEC differed for casein (87.6%), chicken mince (72.6%), heated pea protein concentrate (67.8%), bread (63%), beef entrecote (57.7%) and pea protein concentrate (57.8%). In contrast to existing methods (TCA soluble protein, free NH2-groups), the proposed SEC based method gives separate insight into the two fundamental processes during protein digestion (solubilization and break-down), while maintaining the ability to rank digestibility of very different food proteins.

Advanced structural characterisation of agar-based hydrogels: Rheological and small angle scattering studies.

Agar-based extracts from Gelidium sesquipedale were generated by heat and combined heat-sonication, with and without the application of alkali pre-treatment. Pre-treatment yielded extracts with greater agar contents; however, it produced partial degradation of the agar, reducing its molecular weight. Sonication produced extracts with lower agar contents and decreased molecular weights. A gelation mechanism is proposed based on the rheological and small angle scattering characterization of the extracts. The formation of strong hydrogels upon cooling was caused by the association of agarose chains into double helices and bundles, the sizes of which depended on the agar purity and molecular weight. These different arrangements at the molecular scale consequently affected the mechanical performance of the obtained hydrogels. Heating of the hydrogels produced a gradual disruption of the bundles; weaker or smaller bundles were formed upon subsequent cooling, suggesting that the process was not completely reversible.

Peptides from chicken processing by-product inhibit DPP-IV and promote cellular glucose uptake: potential ingredients for T2D management.

Inhibition of dipeptidyl peptidase IV (DPP-IV) and stimulation of muscle glucose uptake are two of the key strategies for management of type-2-diabetes (T2D). In the present study, four protein hydrolysates generated by enzymatic hydrolysis of chicken by-product, i.e., mechanical chicken deboning residue, were evaluated for their DPP-IV inhibitory activity as well as their effect on glucose uptake by skeletal muscle cells. The DPP-IV inhibitory assay was performed at two concentrations (1000 µg mL-1 and 10 µg mL-1) for the crude chicken protein hydrolysates. The hydrolysate with the highest DPP-IV inhibition was selected for preparative-scale fractionation using size-exclusion chromatography (SEC). The SEC fractions were tested for DPP-IV inhibitory activity as well as their effect on glucose uptake and metabolic activity of skeletal muscle cells. The muscle cells were treated with the SEC fractions and glucose uptake was measured based on luminescence detection of 2-deoxyglucose-6-phosphate (2DG6P). A fraction with peptides in the lower molecular weight range was shown to promote glucose uptake and to inhibit DPP-IV. Further chromatographic fractionation followed by inhibition assaying of the most potent SEC fraction led to isolation of five refined peptide fractions with more than 80% DPP-IV inhibition, which were subsequently analyzed with LC-HRMS/MS. This led to identification of 14 peptides as potential DPP-IV inhibitors from protein hydrolysates of mechanical chicken deboning residue.

At a high dose even partially degraded beta-glucan with decreased solubility significantly reduced the glycaemic response to bread.

Cereal beta-glucan can reduce post-prandial glycaemic responses, which makes it an interesting ingredient to improve the health impact of bread, a staple food with a high glycaemic index (GI). Here we compare the ability of different wheat-based breads prepared with oat bran concentrate and barley flour and a Norwegian type of soft wrap (lompe) for their ability to reduce glycaemic responses in healthy adults. Both breads with the highest beta-glucan content (3.8 g per serving) significantly reduced peak blood glucose rise (PBGR), incremental area under the blood glucose curve (iAUC) and GI compared to wheat control regardless of beta-glucan Mw and solubility. At a medium dose of 1.7 g per serving breads with beta-glucan of high MW and solubility significantly lowered iAUC, but not GI or PBGR compared to white bread. In contrast to previous studies, no significant correlation between viscosity after in vitro digestion and any of the glycaemia variables was found. However, the amount of soluble beta-glucan per serving was inversely correlated with GI. Lompe had a similar medium GI (63) than the high dose beta-glucan breads (56 and 64). However, while "lompe" had significantly lower amounts of rapidly digestible starch, no differences in in vitro starch digestion were found between the different breads. Instead, increased local viscosity at the intestinal border (e.g. soluble beta-glucan interacting with the mucus layer), dilution of nutrients (higher water content and serving size) and/or reduced gastric emptying are proposed as potential explanations for the lower glycaemic responses to high dose beta-glucan breads.

Antifungal activity of well-defined chito-oligosaccharide preparations against medically relevant yeasts.

Due to their antifungal activity, chitosan and its derivatives have potential to be used for treating yeast infections in humans. However, to be considered for use in human medicine, it is necessary to control and know the chemical composition of the compound, which is not always the case for polymeric chitosans. Here, we analyze the antifungal activity of a soluble and well-defined chito-oligosaccharide (CHOS) with an average polymerization degree (DPn) of 32 and fraction of acetylation (FA) of 0.15 (C32) on 52 medically relevant yeast strains. Minimal inhibitory concentrations (MIC) varied widely among yeast species, strains and isolates (from > 5000 to < 9.77 µg mL-1) and inhibition patterns showed a time- and dose-dependencies. The antifungal activity was predominantly fungicidal and was inversely proportional to the pH, being maximal at pH 4.5, the lowest tested pH. Furthermore, antifungal effects of CHOS fractions with varying average molecular weight indicated that those fractions with an intermediate degree of polymerization, i.e. DP 31 and 54, had the strongest inhibitory effects. Confocal imaging showed that C32 adsorbs to the cell surface, with subsequent cell disruption and accumulation of C32 in the cytoplasm. Thus, C32 has potential to be used as a therapy for fungal infections.

Predicting mixed-meal measured glycaemic index in healthy subjects.

PURPOSE: To determine the influence of meal composition on the glycaemic impact of different carbohydrate staples, and the accuracy of "adjusted calculated meal GI" compared with "measured mixed-meal GI". METHODS: In a non-blind randomized crossover trial fasted healthy subjects consumed four dinner-type mixed meals of realistic serving size comprising a carbohydrate staple of either mashed potato, pasta, rice or a glucose drink, combined with fixed portions of boiled carrots, poached salmon and herb sauce. Blood samples collected between 0 and 180 min were analysed for glucose and insulin concentrations. Adjusted calculated meal GI values were determined against a 50 g reference glucose drink, and compared to corresponding measured mixed-meal GIs, supplemented with data from four previous mixed-meal postprandial glycaemic response studies. RESULTS: The common carbohydrate staples, and the glucose drink, ingested as part of the salmon mixed meal induced a significantly lower post-prandial relative glycaemic response (RGR) and concurrent higher relative insulin response than the same amount of staple eaten alone. Adjusted calculated mixed-meal GI closely predicted measured mixed-meal GI in healthy subjects for 15 out of 17 mixed meals examined, showing the need to account for effects of fat and protein when predicting measured mixed-meal GI. Further, we showed the validity of using customarily consumed food amounts in mixed-meal postprandial RGR study design. CONCLUSIONS: Adjusted calculated mixed-meal GI appears a useful model to predict measured mixed-meal GI in healthy subjects and with further development and validation could aid nutrition research and rational design of healthy meals for personalized nutrition and particular consumer groups.

Effects of pH on steam explosion extraction of acetylated galactoglucomannan from Norway spruce.

BACKGROUND: Acetylated galactoglucomannan (AcGGM) is a complex hemicellulose found in softwoods such as Norway spruce (Picea abies). AcGGM has a large potential as a biorefinery feedstock and source of oligosaccharides for high-value industrial applications. Steam explosion is an effective method for extraction of carbohydrates from plant biomass. Increasing the reaction pH reduces the combined severity ( R 0 ' ) of treatment, affecting yields and properties of extracted oligosaccharides. In this study, steam explosion was used to extract oligosaccharides from Norway spruce wood chips soaked with sodium citrate and potassium phosphate buffers with pH of 4.0-7.0. Yields, monosaccharide composition of released oligosaccharides and biomass residue, their acetate content and composition of their lignin fraction were examined to determine the impact of steam explosion buffering on the extraction of softwood hemicellulose. RESULTS: Reducing the severity of steam explosion resulted in lower yields, although the extracted oligosaccharides had a higher degree of polymerization. Higher buffering pH also resulted in a higher fraction of xylan in the extracted oligos. Oligosaccharides extracted in buffers of pH > 5.0 were deacetylated. Buffering leads to a removal of acetylations from both the extracted oligosaccharides and the hemicellulose in the residual biomass. Treatment of the residual biomass with a GH5 family mannanase from Aspergillus nidulans was not able to improve the AcGGM yields. No hydroxymethylfurfural formation, a decomposition product from hexoses, was observed in samples soaked with buffers at pH higher than 4.0. CONCLUSIONS: Buffering the steam explosion reactions proved to be an effective way to reduce the combined severity ( R 0 ' ) and produce a wide range of products from the same feedstock at the same physical conditions. The results highlight the impact of chemical autohydrolysis of hemicellulose by acetic acid released from the biomass in hydrothermal pretreatments. Lower combined severity results in products with a lower degree of acetylation of both the extracted oligosaccharides and residual biomass. Decrease in severity appears not to be the result of reduced acetate release, but rather a result of inhibited autohydrolysis by the released acetate. Based on the results presented, the optimal soaking pH for fine-tuning properties of extracted AcGGM is below 5.0.

Quantification of 1,3-ß-D-glucan from yeast added as a functional ingredient to bread.

Due to their immunomodulatory effect, 1,3-ß-G from yeast are used as functional ingredients, but reliable methods for their detection in foods are lacking. We have adapted a method based on fluorescence detection with aniline blue to quantify the amount of five commercial yeast ß-glucan preparations added to crisp or yeast-leavened bread. This assay detected yeast ß-glucan preparations added to different breads with an average recovery of 90, 96, 99 and 105%, while one of the preparations was overestimated, with an average recovery of 157%. The presence of cereal 1,3-1,4-ß-D-glucans did not interfere with assay performance. The addition of 1,3-ß-G at 0.2 and 0.5 g/100g is low compared to the recommended dose of 1,3-ß-G per serving demonstrating assay sensitivity. However, more research is needed to fully understand the effect of 1,3-ß-G conformation/structure on aniline blue interaction as well as the effect of baking on structure and dissolution properties of yeast ß-glucans.

Glyceamic and insulinaemic response to mashed potato alone, or with broccoli, broccoli fibre or cellulose in healthy adults.

PURPOSE: To examine the role of realistic serving sizes of broccoli, broccoli fibre and cellulose co-consumed with mash potato, or mashed potato eaten alone, on glycaemic and insulinaemic responses (GR and IR) in healthy adults. METHOD: A non-blind randomized crossover trial was conducted with thirteen healthy subjects consuming four different meals. Capillary blood samples between 0 and 180 min were analysed for glucose and insulin. The incremental area under the fasting blood glucose and insulin curves (iAUC) was calculated for different time increments. Differences in GR and IR between meals were assessed by repeated measures analysis of variance. RESULTS: The immediate GR and IR to one serving of mashed potato eaten with two servings of broccoli were significantly lower than mashed potato eaten alone. The peak, incremental peak and iAUC0-30min for GR and iAUC0-30min for IR were all significantly lower for the broccoli-potato meal. This meal also takes longer to return to fasting baseline with a time-delayed lag in IR and GR compared to the potato only meal. The iAUC60-120min for IR was significantly greater for the broccoli-potato meal compared to the other meals. Yet there was no corresponding significant difference between the broccoli-potato meal and the other meals for peak, incremental peak IR or any other iAUCs for GR and IR. For the potato meals containing added broccoli fibre or cellulose, no significant differences in GR or IR were observed when compared with the potato eaten alone. CONCLUSION: Co-consumption of cooked broccoli with mashed potato has a significant effect on glycaemic and insulinaemic responses compared to potato eaten alone. Our study suggests broccoli eaten with potato improves glucose homeostasis and therefore indicates a general beneficial nutritional role for broccoli when eaten with a carbohydrate staple.

Effect of Dietary Fibers on Cecal Microbiota and Intestinal Tumorigenesis in Azoxymethane Treated A/J Min/+ Mice.

Foods naturally high in dietary fiber are generally considered to protect against development of colorectal cancer (CRC). However, the intrinsic effect of dietary fiber on intestinal carcinogenesis is unclear. We used azoxymethane (AOM) treated A/J Min/+ mice, which developed a significantly higher tumor load in the colon than in the small intestine, to compare the effects of dietary inulin (IN), cellulose (CE) or brewers spent grain (BSG) on intestinal tumorigenesis and cecal microbiota. Each fiber was tested at two dose levels, 5% and 15% (w/w) content of the AIN-93M diet. The microbiota was investigated by next-generation sequencing of the 16S rRNA gene (V4). We found that mice fed IN had approximately 50% lower colonic tumor load than mice fed CE or BSG (p<0.001). Surprisingly, all three types of fiber caused a dose dependent increase of colonic tumor load (p<0.001). The small intestinal tumor load was not affected by the dietary fiber interventions. Mice fed IN had a lower bacterial diversity than mice fed CE or BSG. The Bacteroidetes/Firmicutes ratio was significantly (p = 0.003) different between the three fiber diets with a higher mean value in IN fed mice compared with BSG and CE. We also found a relation between microbiota and the colonic tumor load, where many of the operational taxonomic units (OTUs) related to low tumor load were significantly enriched in mice fed IN. Among the OTUs related to low tumor load were bacteria affiliated with the Bacteroides genus. These results suggest that type of dietary fiber may play a role in the development of CRC, and that the suppressive effect of IN on colonic tumorigenesis is associated with profound changes in the cecal microbiota profile.

Towards on-line prediction of dry matter content in whole unpeeled potatoes using near-infrared spectroscopy.

Prediction of dry matter content in whole potatoes is a desired capability in the processing industry. Accurate prediction of dry matter content may greatly reduce waste quantities and improve utilization of the raw material through sorting, hence also reducing the processing cost. The following study demonstrates the use of a low resolution, high speed NIR interactance instrument combined with partial least square regression for prediction of dry matter content in whole unpeeled potatoes. Three different measuring configurations were investigated: (1) off-line measurements with contact between the potato and the light collection tube; (2) off-line measurements without contact between the potato and the light collection tube; and (3) on-line measurements of the potatoes. The offline contact measurements gave a prediction performance of R(2)=0.89 and RMSECV=1.19. Similar prediction performance were obtained from the off-line non-contact measurements (R(2)=0.89, RMSECV=1.23). Significantly better (p=0.038) prediction performance (R(2)=0.92, RMSECV=1.06) was obtained with the on-line measuring configuration, thus showing the possibilities of using the instrument for on-line measurements. In addition it was shown that the dry matter distribution across the individual tuber could be predicted by the model obtained.

Inter-laboratory evaluation of SEC-post-column calcofluor for determination of the weight-average molar mass of cereal ß-glucan.

Even though size exclusion chromatography (SEC) with post column addition of calcofluor (SEC-calcofluor) has been used for the determination of cereal ß-glucan molar mass in foods for many years, there is a lack of systematic evaluation of the method. To address this issue a set of suitable ß-glucan standards were generated by preparative SEC and their molar mass characteristics were determined by analytical multi-detection SEC (refractive index (RI), light scattering). Each standard was then analysed by SEC-calcofluor at three different labs. As a direct comparison, the analyses were repeated with a RI detector. For SEC-calcofluor accurate measurements of weight average molar mass (Mw) can be made for ß-glucan populations within 10-500×10(3)g/mol. Above this molar mass threshold there is an increasing tendency for underestimation of Mw. Precipitation of some ß-glucan-calcofluor complexes may have delayed their transport into the detector.

Viscosity based quantification of endogenous ß-glucanase activity in flour.

High molecular weight (MW) is a key parameter for cereal ß-glucans physiological benefits like decreased serum cholesterol and attenuated post prandial blood glucose. However, the activity of endogenous flour enzymes during bread production results in a decrease of ß-glucan MW. The depolymerization of a standard ß-glucan solution by different flour extracts (wheat, barley and rye) was followed by measuring the viscosity decrease with a Rheometer. The slope of the inverse viscosity (1/η) against degradation time was used to quantify ß-glucanase activity by comparison with slopes obtained with known concentrations of the ß-glucanase Lichenase. Results correlated well with depolymerization rates estimated by HPSEC. The viscosity based method is rapid (20 min per sample), accurate (≤6% variation), and a powerful screening tool for identifying flour fractions with low ß-glucanase activity, treatments that can inactivate ß-glucanases in flour, or the development of ß-glucanase inhibitors for the use in e.g. bread making.

Effects of orally administered yeast-derived beta-glucans: a review.

Yeast-derived beta-glucans (Y-BG) are considered immunomodulatory compounds suggested to enhance the defense against infections and exert anticarcinogenic effects. Specific preparations have received Generally Recognized as Safe status and acceptance as novel food ingredients by European Food Safety Authority. In human trials, orally administered Y-BG significantly reduced the incidence of upper respiratory tract infections in individuals susceptible to upper respiratory tract infections, whereas significant differences were not seen in healthy individuals. Increased salivary IgA in healthy individuals, increased IL-10 levels in obese subjects, beneficial changes in immunological parameters in allergic patients, and activated monocytes in cancer patients have been reported following Y-BG intake. The studies were conducted with different doses (7.5-1500 mg/day), using different preparations that vary in their primary structure, molecular weight, and solubility. In animal models, oral Y-BG have reduced the incidence of bacterial infections and levels of stress-induced cytokines and enhanced antineoplastic effects of cytotoxic agents. Protective effects toward drug intoxication and ischemia/reperfusion injury have also been reported. In conclusion, additional studies following good clinical practice principles are needed in which well-defined Y-BG preparations are used and immune markers and disease endpoints are assessed. Since optimal dosing may depend on preparation characteristics, dose-response curves might be assessed to find the optimal dose for a specific preparation.

Generic tools to assess genuine carbohydrate specific effects on in vitro immune modulation exemplified by ß-glucans.

Even if carbohydrate preparations from plant/fungal sources have a high degree of purity, observed immune-stimulation may be caused by minute sample contaminations. Using the example of different ß-glucans we present a range of analytical tools crucial for validation of possible immune-stimulatory effects. Two yeast (MacroGard and Zymosan) and one cereal ß-glucan (CBG40) increased IL-8 secretion by HT-29 cells considerably. Degradation of the ß-glucan samples with ß-glucan specific enzymes did hardly influence the effect of Zymosan and CBG40 but significantly decreased the effect of MacroGard. Stimulation of IL-8 secretion by CBG40 and Zymosan was hence not due to their ß-glucan content. Instead, the effect of the CBG40 sample was due to low levels of LPS despite the inability of the known LPS inhibitor Polymyxin B to supress its stimulatory effect. We conclude that targeted enzymatic degradation of samples is a powerful validation tool to investigate carbohydrate specific immune-modulation.

Cereal ß-glucan quantification with calcofluor-application to cell culture supernatants.

The specific binding of the fluorescent dye calcofluor to cereal ß-glucan results in increased fluorescence intensity of the formed complex and is in use for the quantification of ß-glucan above a critical molecular weight (MW) by flow injection analysis. In this study, this method was applied in a fast and easy batch mode. In order to emphasize the spectral information of the emission spectra of the calcofluor/ß-glucan complexes, derivative signals were calculated. A linear relationship was found between the amplitude of the second derivative signals and the ß-glucan concentration between 0.1 and 0.4 µg/mL. The low detection limit of this new method (0.045 µg/mL) enabled its use to study the transport of cereal ß-glucans over differentiated Caco-2 cell monolayers. Additionally, the method was applied to quantify ß-glucan in arabinoxylan samples, which correlated well with data by an enzyme based method.

Application of high-performance anion-exchange chromatography with pulsed amperometric detection to compare the kinetic properties of ß-glucosidase on oligosaccharides from lichenase digested barley ß-glucan.

A chromatographic method using HPAEC-PAD was developed to accurately quantify the major oligosaccharides derived from lichenase degradation of barley ß-glucan. This method was further used to follow ß-glucosidase degradation and product formation as progress curves. This approach allowed us to compare the kinetic characteristics of ß-glucosidase on each exclusive oligosaccharide substrate and their mixtures. Our results show that when determining the kinetic parameters of exohydrolases on oligosaccharides following the progress curve for the substrates is necessary since calculations based only on released monosaccharide products may lead to an error. The catalytic activity of almond ß-glucosidase on laminaribiose (G3G) was approximately half that measured against cellobiose (3.15 S(-1)). The enzyme had 12 times and 560 times less catalytic activity on 3-O-ß-cellobiosyl-D-glucose (G4G3G) and on 3-O-ß-cellotriosyl-d-glucose (G4G4G3G) respectively than on G3G. Our approach offers a useful tool for the determination of the kinetics of enzymatic or chemical modification of various carbohydrate substrates.