Effect of Bioprocessing on Techno-Functional Properties of Climate-Resilient African Crops, Sorghum and Cowpea.

Sorghum and cowpea are very compatible for intercropping in hot and dry environments, and they also have complementary nutritional compositions. Thus, the crops have the potential to improve food security in regions threatened by climate change. The aim of this study was to investigate different enzymes (carbohydrate-degrading, proteases and phytases) and lactic acid bacteria (LAB) fermentation to improve the techno-functional properties of sorghum and cowpea flours. Results show that sorghum carbohydrates were very resistant to hydrolysis induced by bioprocessing treatments. Most of the protease treatments resulted in low or moderate protein solubilization (from ca. 6.5% to 10%) in sorghum, while the pH adjustment to 8 followed by alkaline protease increased solubility to 40%. With cowpea, protease treatment combined with carbohydrate-degrading enzymes increased the solubility of proteins from 37% up to 61%. With regard to the techno-functional properties, LAB and amylase treatment decreased the sorghum peak paste viscosities (from 504 to 370 and 325 cPa, respectively), while LAB and chemical acidification increased cowpea viscosity (from 282 to 366 and 468 cPa, respectively). When the bioprocessed sorghum and cowpea were tested in breadmaking, only moderate effects were observed, suggesting that the modifications by enzymes and fermentation were not strong enough to improve breadmaking.

Impact of Enzymatic Hydrolysis and Microfluidization on the Techno-Functionality of Oat Bran in Suspension and Acid Milk Gel Models.

Oat bran is a nutritionally rich ingredient, but it is underutilized in semi-moist and liquid foods due to technological issues such as high viscosity and sliminess. The aim of this work was to improve the technological properties of oat bran concentrate (OBC) in high-moisture food applications by enzymatic and mechanical treatments. OBC was hydrolyzed with ß-glucanase (OBC-Hyd) and the water-soluble fraction (OBC-Sol) was separated. OBC, OBC-Hyd and OBC-Sol were further microfluidized at 5% dry matter content. Enzymatic treatment and microfluidization of OBC reduced the molecular weight (Mw) of ß-glucan from 2748 kDa to 893 and 350 kDa, respectively, as well as the average particle size of OBC (3.4 and 35 times, respectively). Both treatments increased the extractability of the soluble compounds from the OBC samples (up to 80%) and affected their water retention capacity. OBC in suspension had very high viscosity (969 mPa·s) when heated, which decreased after both enzyme and microfluidization treatments. The colloidal stability of the OBC in suspension was improved, especially after microfluidization. The addition of OBC samples to acid milk gels decreased syneresis, improved the water holding capacity and softened the texture. The changes in the suspension and gel characteristics were linked with reduced ß-glucan Mw and OBC particle size.

Role of ß-glucan content, molecular weight and phytate in the bile acid binding of oat ß-glucan.

There is controversy about the role of viscosity and co-migrating molecules on the bile acid binding of beta-glucan. Thus, this study aimed to investigate the impact of ß-glucan molecular weight and the content of both ß-glucan and phytate on the mobility of bile acids by modelling intestinal conditions in vitro. Two approaches were used to evaluate factors underlying this binding effect. The first studied bile acid binding capacity of soluble ß-glucan using purified compounds. Viscosity of the ß-glucan solution governed mainly the mobility of bile acid since both a decrease in ß-glucan concentration and degradation of ß-glucan by enzyme hydrolysis resulted in decreased binding. The second approach investigated the trapping of bile acids in the oat bran matrix. Results suggested trapping of bile acids by the ß-glucan gel network. Additionally, hydrolysis of phytate was shown to increase bile acid binding, probably due to better extractability of ß-glucan in this sample.

Effect of oat ß-glucan of different molecular weights on fecal bile acids, urine metabolites and pressure in the digestive tract - A human cross over trial.

While the development of oat products often requires altered molecular weight (MW) of ß-glucan, the resulting health implications are currently unclear. This 3-leg crossover trial (n = 14) investigated the effects of the consumption of oat bran with High, Medium and Low MW ß-glucan (average > 1000, 524 and 82 kDa respectively) with 3 consequent meals on oat-derived phenolic compounds in urine (UHPLC-MS/MS), bile acids in feces (UHPLC-QTOF), gastrointestinal conditions (ingestible capsule), and perceived gut well-being. Urine excretion of ferulic acid was higher (p < 0.001, p < 0.001), and the fecal excretion of deoxycholic (p < 0.03, p < 0.02) and chenodeoxycholic (p < 0.06, p < 0.02) acids lower after consumption of Low MW ß-glucan compared with both Medium and High MW ß-glucan. Duodenal pressure was higher after consumption of High MW ß-glucan compared to Medium (p < 0.041) and Low (p < 0.022) MW ß-glucan. The MW of ß-glucan did not affect gut well-being, but the perceptions between females and males differed.

Effect of Varying Molecular Weight of Oat ß-Glucan Taken just before Eating on Postprandial Glycemic Response in Healthy Humans.

To see if the molecular weight (MW) and viscosity of oat ß-glucan (OBG) when taken before eating determine its effect on postprandial glycemic responses (PPRG), healthy overnight-fasted subjects (n = 16) were studied on eight separate occasions. Subjects consumed 200 mL water alone (Control) or with 4 g OBG varying in MW and viscosity followed, 2-3 min later, by 113 g white-bread. Blood was taken fasting and at 15, 30, 45, 60, 90, and 120 min after starting to eat. None of the OBG treatments differed significantly from the Control for the a-priori primary endpoint of glucose peak-rise or secondary endpoint of incremental area-under-the-curve (iAUC) over 0-120 min. However, significant differences from the Control were seen for glucose iAUC over 0-45 min and time to peak (TTP) glucose. Lower log(MW) and log(viscosity) were associated with higher iAUC 0-45 (p < 0.001) and shorter TTP (p < 0.001). We conclude that when 4 g OBG is taken as a preload, reducing MW does not affect glucose peak rise or iAUC0-120, but rather accelerates the rise in blood glucose and reduces the time it takes glucose to reach the peak. However, this is based on post-hoc calculation of iAUC0-45 and TTP and needs to be confirmed in a subsequent study.

In vitro study for investigating the impact of decreasing the molecular weight of oat bran dietary fibre components on the behaviour in small and large intestine.

The objective of this work was to evaluate the role of ß-glucan molecular weight (Mw) and the presence of other carbohydrates on the physiological functionality of oat bran via an in vitro digestion study. A complete approach using three different in vitro digestion models (viscosity of the small intestine digest, reduction of bile acids and on-line measurement of gas evolution) was used to predict the physiological functionality of enzymatically modified oat bran concentrate (OBC). OBC was enzymatically treated with two ß-glucanase preparations at three different levels in order to specifically decrease ß-glucan Mw (Pure: purified ß-glucanase) or ß-glucan and other cell wall polysaccharides (Mix: commercial food-grade cell wall degrading enzyme preparation). The Mw of ß-glucan in OBC was tailored to high (1000 kDa), medium (200-500 kDa) and low (<100 kDa) values. The amount of arabinoxylan-oligosaccharides varied from 0.3 to 4.7 g per 100 g of OBC when OBC was treated with the Mix enzyme at the highest dosage. When the enzymatically treated OBCs were studied in an upper gut model, a decrease in the viscosity of the digest simultaneously with the reduction of ß-glucan Mw was observed. At a similar ß-glucan Mw range, OBC samples treated with the Pure enzyme had lower viscosity than the samples treated with the Mix one, which also contained arabinoxylan-oligosaccharides. After enzymatic hydrolysis, the capacity of OBC to reduce bile acid was decreased regardless of the enzyme treatment used, and a positive correlation was found between ß-glucan Mw and bile acid reduction (r = 0.99**). The production of colonic gases by the enzymatically treated OBC samples in an in vitro colon model showed an inverse correlation between ß-glucan Mw and initial rate of gas formation (r = -0.9**), but no impact of arabinoxylan-oligosaccharides was observed. This study emphasised the complexity of factors affecting the functionality of oat components under physiological conditions and demonstrated the possibility to produce Mw-tailored oat fibre ingredients that could contribute to gut mediated health benefits.

Study into the effect of microfluidisation processing parameters on the physicochemical properties of wheat (Triticum aestivum L.) bran.

The physicochemical properties of wheat bran have an effect on its technofunctional and nutritional profile. The possibility to induce physicochemical modifications in wheat bran using microfluidisation was investigated. An I-optimal experimental design was used to investigate the effect of microfluidisation processing parameters (pressure, number of passes, bran concentration and initial particle size) on important properties of wheat bran (particle size, microstructure, chemical composition, water retention capacity (WRC), extractability, viscosity and sedimentation). With the parameters used in this study, microfluidisation reduced wheat bran median particle size to 14.8 µm and disintegrated starch granules from the attached endosperm. This coincided with an increased extractability of starch and arabinoxylan. While the initial particle size was of minor importance, a higher pressure, larger number of passes and lower bran concentration during microfluidisation resulted in a smaller particle size, higher WRC and extractability, and an increased viscosity and stability in a 2% wheat bran suspension.

Contribution of gut microbiota to metabolism of dietary glycine betaine in mice and in vitro colonic fermentation.

BACKGROUND: Accumulating evidence is supporting the protective effect of whole grains against several chronic diseases. Simultaneously, our knowledge is increasing on the impact of gut microbiota on our health and on how diet can modify the composition of our bacterial cohabitants. Herein, we studied C57BL/6 J mice fed with diets enriched with rye bran and wheat aleurone, conventional and germ-free C57BL/6NTac mice on a basal diet, and the colonic fermentation of rye bran in an in vitro model of the human gastrointestinal system. We performed 16S rRNA gene sequencing and metabolomics on the study samples to determine the effect of bran-enriched diets on the gut microbial composition and the potential contribution of microbiota to the metabolism of a novel group of betainized compounds. RESULTS: The bran-enriched study diets elevated the levels of betainized compounds in the colon contents of C57BL/6 J mice. The composition of microbiota changed, and the bran-enriched diets induced an increase in the relative abundance of several bacterial taxa, including Akkermansia, Bifidobacterium, Coriobacteriaceae, Lactobacillus, Parasutterella, and Ruminococcus, many of which are associated with improved health status or the metabolism of plant-based molecules. The levels of betainized compounds in the gut tissues of germ-free mice were significantly lower compared to conventional mice. In the in vitro model of the human gut, the production of betainized compounds was observed throughout the incubation, while the levels of glycine betaine decreased. In cereal samples, only low levels or trace amounts of other betaines than glycine betaine were observed. CONCLUSIONS: Our findings provide evidence that the bacterial taxa increased in relative abundance by the bran-based diet are also involved in the metabolism of glycine betaine into other betainized compounds, adding another potential compound group acting as a mediator of the synergistic metabolic effect of diet and colonic microbiota.

Decreased plasma serotonin and other metabolite changes in healthy adults after consumption of wholegrain rye: an untargeted metabolomics study.

BACKGROUND: Wholegrain consumption has been associated with beneficial health effects including reduction of diabetes and cancer risk; however, the underlying mechanisms are not fully understood. OBJECTIVE: The aim of this study was to characterize the effects of wholegrain rye intake on circulating metabolites in a human intervention study using untargeted metabolomics. METHODS: The intervention consisted of 2 successive 4-wk periods in a randomized crossover design, where 15 adults consumed wholegrain rye bread (WGR) or white wheat bread enriched with fermented rye bran (WW+RB), following a 4-wk rye-free period with white wheat bread (WW). Fasting plasma samples were collected at the end of each period and analyzed using liquid chromatography-mass spectrometry. Metabolic profiles were compared to identify compounds discriminating WGR from the WW+RB and WW periods. Because peripheral serotonin is produced mainly in the gut, a hypothesis of its altered biosynthesis as a response to increased cereal fiber intake was tested by measuring intestinal serotonin of mice fed for 9 wk on a high-fat diet supplemented with different sources of fiber (rye bran flour, ground wheat aleurone, or powdered cellulose). RESULTS: Five endogenous metabolites and 15 rye phytochemicals associated with WGR intake were identified. Plasma concentrations of serotonin, taurine, and glycerophosphocholine were significantly lower after the WGR than WW period (Q < 0.05). Concentrations of 2 phosphatidylethanolamine plasmalogens, PE(18:2/P-18:0) and PE(18:2/P-16:0), were lower after the WGR period than the WW+RB period (Q < 0.05). The concentration of serotonin was significantly lower in the colonic tissue of mice that consumed rye bran or wheat aleurone compared with cellulose (P < 0.001). CONCLUSIONS: Wholegrain rye intake decreases plasma serotonin in healthy adults when compared with refined wheat. Intake of rye bran and wheat aleurone decreases colonic serotonin in mice. These results suggest that peripheral serotonin could be a potential link between wholegrain consumption and its associated health effects.Data used in the study were derived from a trial registered at www.clinicaltrials.gov as NCT03550365.

A Small In Vitro Fermentation Model for Screening the Gut Microbiota Effects of Different Fiber Preparations.

The development of prebiotic fibers requires fast high-throughput screening of their effects on the gut microbiota. We demonstrated the applicability of a mictotiter plate in the in vitro fermentation models for the screening of potentially-prebiotic dietary fibers. The effects of seven rye bran-, oat- and linseed-derived fiber preparations on the human fecal microbiota composition and short-chain fatty acid production were studied. The model was also used to study whether fibers can alleviate the harmful effects of amoxicillin-clavulanate on the microbiota. The antibiotic induced a shift in the bacterial community in the absence of fibers by decreasing the relative amounts of Bifidobacteriaceae, Bacteroidaceae, Prevotellaceae, Lachnospiraceae and Ruminococcaceae, and increasing proteobacterial Sutterilaceae levels from 1% to 11% of the total microbiota. The fermentation of rye bran, enzymatically treated rye bran, its insoluble fraction, soluble oat fiber and a mixture of rye fiber:soluble oat fiber:linseed resulted in a significant increase in butyrate production and a bifidogenic effect in the absence of the antibiotic. These fibers were also able to counteract the negative effects of the antibiotic and prevent the decrease in the relative amount of bifidobacteria. Insoluble and soluble rye bran fractions and soluble oat fiber were the best for controlling the level of proteobacteria at the level below 2%.

Phytic Acid Reduction by Bioprocessing as a Tool To Improve the In Vitro Digestibility of Faba Bean Protein.

In legumes such as faba bean, phytic acid can form very stable complexes with proteins, thus hindering their accessibility for gastrointestinal digestion. This study was set up to investigate the influence of enzymatic phytase treatment and lactic acid bacteria fermentation ( Lactobacillus plantarum VTT E-78076) on phytic acid reduction as well as on the protein quality and digestibility of faba bean flour. The soluble protein and release of free amino acids from faba bean flours were followed during a simulated in vitro digestion process. Phytase treatment efficiently reduced the amount of phytic acid in faba bean flour up to 89% degradation. The reduction of phytic acid content caused a shift in the protein solubility curve, showing higher solubility levels at low pH. Further, the enzyme-aided degradation of phytic acid considerably enhanced the digestibility of faba bean proteins (dose-dependent) and the release of free amino nitrogen in the first stage of digestion (gastric phase). The results propose the benefits of phytase treatment in plant-based foods where the phytic acid content may hinder the digestion of protein.

Diets rich in whole grains increase betainized compounds associated with glucose metabolism.

Background: Epidemiologic evidence suggests that diets rich in whole grains are associated with a reduced risk of developing chronic diseases and all-cause mortality. However, the molecular mechanisms behind these beneficial metabolic effects are poorly understood. Objective: Our aim was to investigate novel trimethylated (betainized) compounds from mice and humans, and their association with whole grain-rich diets and insulin resistance and insulin secretion. Design: Fasting plasma samples were obtained in a mouse (C57BL/6J male) feeding trial and a controlled dietary intervention. The mouse trial involved feeding the mice a rye and wheat bran-enriched feed which was compared with a high-fat diet. In the human trial, participants recruited from Kuopio, Finland (n = 69) and Naples, Italy (n = 54) with characteristics of the metabolic syndrome were randomly assigned to either a whole grain-enriched diet or a control diet for 12 wk. Plasma concentrations of betainized compounds were analyzed with the use of liquid chromatography-tandem mass spectrometry. Insulin resistance and insulin secretion were assessed in an oral-glucose-tolerance test and a meal-glucose-tolerance test. Results: The betaines that were increased in mouse plasma after bran-enriched feeding were identified de novo via chemical synthesis and liquid chromatography-tandem mass spectrometry, and confirmed to be associated with an increased intake of whole-grain products in humans. In particular, the concentrations of pipecolic acid betaine were increased at the end of the whole-grain intervention in both the Kuopio cohort (P < 0.001) and the Naples cohort (P < 0.05), and these concentrations inversely correlated with the postprandial glucose concentration. Furthermore, the concentration of valine betaine was substantially increased during the intervention in Naples (P < 0.001) with an inverse correlation with the postprandial insulin concentration. In addition, the concentrations of other betaines, e.g., glycine betaine and proline betaine, correlated with glucose and insulin concentrations at the end of the intervention. Conclusions: Novel betainized compounds in humans are associated with diets rich in whole grains, and they improve insulin resistance and insulin secretion. These results suggest that these novel compounds may contribute to the beneficial effects of whole grain-rich diets. The studies were registered at clinicaltrials.gov as NCT00945854 (Naples) and NCT00573781 (Kuopio).

Structural properties and foaming of plant cell wall polysaccharide dispersions.

Water suspensions of cellulose nanofibres with xylan, xyloglucan and pectin were studied for foaming and structural properties as a new means for food structuring. The dispersions were analysed with rheological measurements, microscopy and optical coherence tomography. A combination of xylan with TEMPO-oxidized nanocellulose produced a mixture with well-dispersed air bubbles, while the addition of pectin improved the elastic modulus, hardness and toughness of the structures. A similar structure was observed with native nanocellulose, but the elastic modulus was not as high. Shear flow caused cellulose nanofibres to form plate-like flocs in the suspension that accumulated near bubble interfaces. This tendency could be affected by adding laccase to the dispersion, but the effect was opposite for native and TEMPO-oxidized nanocellulose. Nanocellulose type also influenced the interactions between nanofibers and other polysaccharides. For example, xyloglucan interacted strongly with TEMPO-oxidized nanocellulose (high storage modulus) but not with native nanocellulose.

Birch pulp xylan works as a food hydrocolloid in acid milk gels and is fermented slowly in vitro.

The objective was to evaluate the potential of birch xylan as a food hydrocolloid and dietary fibre. High-molecular weight xylan was isolated from birch kraft pulp by alkaline extraction, and enzymatically hydrolysed. Fermentability of xylans was evaluated using an in vitro colon model and performance as a hydrocolloid was studied in low-fat acid milk gels (1.5% and 3% w/w). Texture of the gels and water holding capacity of xylans were compared with inulin, fructooligosaccharide and xylooligosaccharide. Xylans showed slower fermentation rate by faecal microbiota than the references. Xylan-enriched acid milk gels (3% w/w) had improved water holding capacity (over 2-fold) and showed lower spontaneous syneresis, firmness and elasticity when compared to control (no hydrocolloids) or to references. In conclusion, birch xylan improved texture of low-fat acid milk gel applications, and the slow in vitro fermentation rate predicts lower incidence of intestinal discomfort in comparison to the commercial references.

Amino acid-derived betaines dominate as urinary markers for rye bran intake in mice fed high-fat diet--A nontargeted metabolomics study.

SCOPE: Bioprocessing of whole grain cereals may affect the bioavailability of phytochemicals associated with grain fiber and ultimately lead to different health outcomes. Here, we studied the impact of long-term feeding with intact and bioprocessed rye bran on the urinary phytochemical profile of mice. METHODS AND RESULTS: Nontargeted hydrophilic interaction chromatography-ESI-qTOF-MS metabolite profiling approach was applied on urine samples collected from three groups of diet-induced obese mice fed for 8 weeks with one of the three diets: high-fat (HF) control diet, HF diet enriched with intact rye bran, or HF diet enriched with bioprocessed rye bran. The most striking finding was the increased urinary excretion of several amino-acid derived betaines after both rye diets. These included proline betaine, alanine betaine, valine betaine, phenylalanine betaine, pipecolic acid betaine, and trigonelline, but not glycine betaine. Furthermore, bioprocessing may have improved the bioavailability of rye-derived phytochemicals, as higher increase in, e.g. ferulic acid and benzoxazinoid metabolites were observed in urine of mice fed with bioprocessed than intact rye bran. CONCLUSION: Urinary excretion of various betaines was greatly increased in mice fed rye brans. Furthermore, bioprocessing of rye bran appears to serve as a beneficial way to improve the bioavailability of various phytochemicals.