Brewers' spent grain as substrate for dextran biosynthesis by Leuconostoc pseudomesenteroides DSM20193 and Weissella confusa A16.

BACKGROUND: Lactic acid bacteria can synthesize dextran and oligosaccharides with different functionality, depending on the strain and fermentation conditions. As natural structure-forming agent, dextran has proven useful as food additive, improving the properties of several raw materials with poor technological quality, such as cereal by-products, fiber-and protein-rich matrices, enabling their use in food applications. In this study, we assessed dextran biosynthesis in situ during fermentation of brewers´ spent grain (BSG), the main by-product of beer brewing industry, with Leuconostoc pseudomesenteroides DSM20193 and Weissella confusa A16. The starters performance and the primary metabolites formed during 24 h of fermentation with and without 4% sucrose (w/w) were followed. RESULTS: The starters showed similar growth and acidification kinetics, but different sugar utilization, especially in presence of sucrose. Viscosity increase in fermented BSG containing sucrose occurred first after 10 h, and it kept increasing until 24 h concomitantly with dextran formation. Dextran content after 24 h was approximately 1% on the total weight of the BSG. Oligosaccharides with different degree of polymerization were formed together with dextran from 10 to 24 h. Three dextransucrase genes were identified in L. pseudomesenteroides DSM20193, one of which was significantly upregulated and remained active throughout the fermentation time. One dextransucrase gene was identified in W. confusa A16 also showing a typical induction profile, with highest upregulation at 10 h. CONCLUSIONS: Selected lactic acid bacteria starters produced significant amount of dextran in brewers' spent grain while forming oligosaccharides with different degree of polymerization. Putative dextransucrase genes identified in the starters showed a typical induction profile. Formation of dextran and oligosaccharides in BSG during lactic acid bacteria fermentation can be tailored to achieve specific technological properties of this raw material, contributing to its reintegration into the food chain.

How dietary intake has been assessed in African countries? A systematic review.

BACKGROUND: Dietary patterns are often considered as one of the main causes of non-communicable diseases worldwide. It is of utmost importance to study dietary habits in developing countries since this work is scarce. OBJECTIVE: To summarize the most recent research conducted in this field in African countries, namely the most used methodologies and tools. METHODS: A systematic review was conducted on MEDLINE®/PubMed, aiming to identify scientific publications focused on studies of dietary intake of different African populations, in a ten-year period. Papers not written in English/Portuguese/Spanish, studies developed among African people but not developed in African countries, studies aiming to assess a particular nutrient/specific food/food toxin and studies that assessed dietary intake among children were excluded. FINDINGS: Out of 99 included studies, the 24-hour recall and the food-frequency questionnaire were the most used dietary intake assessment tools, used to assess diet at an individual level. It was also observed that often country-unspecific food composition databases are used, and the methodologies employed are poorly validated and standardized. CONCLUSIONS: There is an emergent need to improve the existing food databases by updating food data and to develop suitable country-specific databases for those that do not have their own food composition table.

Rye bran as fermentation matrix boosts in situ dextran production by Weissella confusa compared to wheat bran.

The consumption of fiber-rich foods such as cereal bran is highly recommended due to its beneficial health effects. Pre-fermentation of bran with lactic acid bacteria can be used to improve the otherwise impaired flavor and textural qualities of bran-rich products. These positive effects are attributed to enzymatic modification of bran components and the production of functional metabolites like organic acids and exopolysaccharides such as dextrans. The aim of this study was to investigate dextran production in wheat and rye bran by fermentation with two Weissella confusa strains. Bran raw materials were analyzed for their chemical compositions and mineral content. Microbial growth and acidification kinetics were determined from the fermentations. Both strains produced more dextran in rye bran in which the fermentation-induced acidification was slower and the acidification lag phase longer than in wheat bran. Higher dextran production in rye bran is expected to be due to the longer period of optimal pH for dextran synthesis during fermentation. The starch content of wheat bran was higher, which may promote isomaltooligosaccharide formation at the expense of dextran production. W. confusa Cab3 produced slightly higher amounts of dextran than W. confusa VTT E-90392 in all raw materials. Fermentation with W. confusa Cab3 also resulted in lower residual fructose content which has technological relevance. The results indicate that wheat and particularly rye bran are promising matrices for producing technologically significant amounts of dextran, which facilitates the use of nutritionally valuable raw bran in food applications.

Characterization of exopolysaccharide and ropy capsular polysaccharide formation by Weissella.

With their broad functional properties, lactic acid bacteria derived high molar mass exopolysaccharides (EPS) and oligosaccharides are of great interest for food, medical and pharmaceutical industry. EPS formation by 123 strains of Weissella cibaria and Weissella confusa, was evaluated. Dextran formation from sucrose was observed for all tested strains while 18 strains produced fructan in addition to dextran. Six isolates synthesized a highly ropy polymer from glucose associated with the formation of a cell-bound, capsular polysaccharide (CPS) composed of glucose, O-acetyl groups and two unidentified monomer components. The soluble EPSs of nine strains were identified as low α-1,3-branched dextran, levan and inulin type polymers using NMR. In addition to glucan and fructan, W. confusa produced gluco- and fructooligosaccharides. Partial dextransucrase and fructansucrase sequences were characterized in the selected Weissella strains. Our study reports the first structural characterization of fructan type EPS from Weissella as well as the first Weissella strain producing inulin. Production of more than one EPS-type by single strains may have high potential for development of applications combining EPS technological and nutritional benefits.

Production and sensory evaluation of composite breads based on wheat and whole millet or sorghum in the presence of Weissella confusa A16 exopolysaccharides.

The aim of the study was to evaluate the acceptability of composite breads based on local cereal (millet and sorghum) formulations. Bread preparations based on 50% wheat flour and 50% local cereal flour were made in the presence of exopolysaccharide (Eps) production stimulated by a strain of Weissella confusa A16 in the fermented dough. Seven formulations were done in two baking sets and were submitted to sensory evaluations which consisted of tests on sensory profile, hedonic analysis and ranking. Results showed that the presence of Eps improved the acceptability of breads made with local cereal flours. The white color of the crumb of breads made with 100% wheat flour was the most appreciated by consumers. The less local flour is used in the bread preparation, the better the bread is appreciated. Nevertheless, formulations containing whole grains were the least appreciated, partly because of the hardness of the breads. Interestingly, more than 50% of consumers found the taste pleasant for breads made with 50% millet flour.

Nutritional intake and food sources in an adult urban Kenyan population.

Urbanisation is hastening the transition from traditional food habits to less healthy diets, which are becoming more common among Kenyans. No up-to-date studies on usual dietary intake and the main food sources of adult Kenyans are available. The aim of the present study was to identify the main food sources of nutrients in the diet of urban adult Kenyans and explore potential associations with demographic variables including age, sex, level of education, occupation and body mass index. The study adopted a cross-sectional design. The dietary intake of 486 adult Kenyans from Nairobi was assessed using a validated, culture-sensitive, semi-quantitative food frequency questionnaire. Binary logistic regression models were used to evaluate associations between food sources and demographic variables. Macronutrient intakes as a proportion of total energy intake (TEI) were within international dietary guidelines. Cereals and grain products (34.0%), sugar, syrups, sweets and snacks (9.8%), fruits (9.7%) and meat and eggs (8.8%) were the major contributors to TEI. Cereals and grain products contributed 42.5% to carbohydrates, followed by fruits (12.4%) and sugar, syrups, sweets and snacks (10.6%). The most important sources of protein and total fat were cereals and grain products (23.3% and 19.7%, respectively) and meat and eggs (22.0% and 18.7%, respectively). Sex, age and level of education were associated with the choice of food groups. Although macronutrient intakes were within guidelines, the Kenyan diet was revealed to be high in sugars, salt and fibre, with differences in food sources according to demographic variables. These results can act as an incentive to national authorities to implement nutritional strategies aiming to raise awareness of healthier dietary patterns among Kenyans.

Structural analysis of enzyme-resistant isomaltooligosaccharides reveals the elongation of α-(1→3)-linked branches in Weissella confusa dextran.

Weissella confusa VTT E-90392 is an efficient producer of a dextran that is mainly composed of α-(1→6)-linked D-glucosyl units and very few α-(1→3) branch linkages. A mixture of the Chaetomium erraticum endodextranase and the Aspergillus niger α-glucosidase was used to hydrolyze W. confusa dextran to glucose and a set of enzyme-resistant isomaltooligosaccharides. Two of the oligosaccharides (tetra- and hexasaccharide) were isolated in pure form and their structures elucidated. The tetrasaccharide had a nonreducing end terminal α-(1→3)-linked glucosyl unit (α-D-Glcp-(1→3)-α-D-Glcp-(1→6)-α-D-Glcp-(1→6)-α-D-Glc), whereas the hexasaccharide had an α-(1→3)-linked isomaltosyl side group (α-D-Glcp-(1→6)[α-D-Glcp-(1→6)-α-D-Glcp-(1→3)]-α-D-Glcp-(1→6)-α-D-Glcp-(1→6)-α-D-Glc). A mixture of two isomeric oligosaccharides was also obtained in the pentasaccharide fraction, which were identified as (α-D-Glcp-(1→6)-α-D-Glcp-(1→3)-α-D-Glcp-(1→6)-α-D-Glcp-(1→6)-α-D-Glc) and (α-D-Glcp-(1→6)[α-D-Glcp-(1→3)]-α-D-Glcp-(1→6)-α-D-Glcp-(1→6)-α-D-Glc). The structures of the oligosaccharides indicated that W. confusa dextran contains both terminal and elongated α-(1→3)-branches. This is the first report evidencing the presence of elongated branches in W. confusa dextran. The (1)H and (13)C NMR spectroscopic data on the enzyme-resistant isomaltooligosaccharides with α-(1→3)-linked glucosyl and isomaltosyl groups are published here for the first time.

Isolation and characterization of indigenous Weissella confusa for in situ bacterial exopolysaccharides (EPS) production in chickpea sourdough.

Legume-based sourdough represents a potential ingredient for the manufacture of novel baked products. However, the lack of gluten of legume flours can restrict their use due to their poor technological properties. To overcome such issue, the in situ production of bacterial exopolysaccharides (EPS) during fermentation has been proposed. In this study, an EPS-producing lactic acid bacteria for in situ production in chickpea sourdough was isolated. After several backsloppings of the spontaneously fermented chickpea flour dough, a dominant strain of Weissella confusa was isolated and identified. W. confusa Ck15 was able to produce linear dextran with 2.6% α-(1 â†’ 3) linked branches, from sucrose. Temperature of 30 °C, dough yield of 333, and 2% of sucrose addition were used to produce fermented chickpea sourdoughs. The acidification and rheology of the sourdoughs inoculated with W. confusa Ck15, Leuconostoc pseudomesenteroides DSM 20193, as positive control, and Lactobacillus plantarum F8, as negative control, were compared. The in situ dextran production by W. confusa Ck15 fermentation led to the highest viscosity increase (5.90 Pa·s) and the highest EPS percentage in the doughs (1.49%), compared to the other doughs. The in situ dextran production represents a potential approach for improving the use of legume flour in bakery products; overall, this experiment represents a first step for the exploitation of microbial EPS for setting up a baking process for chickpea based product.

In situ production and analysis of Weissella confusa dextran in wheat sourdough.

Several lactic acid bacteria belonging to the genera Leuconostoc, Lactobacillus, and Weissella have been introduced to wheat sourdough baking for in situ production of exopolysaccharides. This is considered a novel method for improving the shelf-life, volume and nutritional value of bread without additives. However, in situ production of exopolysaccharides during sourdough fermentation is challenged by simultaneous acidification due to metabolic activities of the bacteria, which may significantly diminish the positive technological impact of exopolysaccharides. In this study, the growth, activity and in situ production of dextran by Weissella confusa VTT E-90392 in wheat sourdoughs were investigated. Furthermore, the influence of dextran-enriched sourdoughs, at the addition level of 43%, on the subsequent bread quality was established. W. confusa efficiently produced dextran from the added sucrose in wheat sourdough without strong acid production. A new specific enzyme-assisted method for in situ analysis of dextran in sourdoughs was developed. With this method, we could for the first time proof significant (11-16 g/kg DW) production of polymeric dextran in sourdoughs. Concomitant formation of shorter isomaltooligosaccharides by W. confusa was also detected. The produced dextran significantly increased the viscosity of the sourdoughs. Application of dextran-enriched sourdoughs in bread baking provided mildly acidic wheat bread with improved volume (up to 10%) and crumb softness (25-40%) during 6 days of storage. Hence, W. confusa is a promising new strain for efficient in situ production of dextrans and isomaltooligosaccharides in sourdoughs without strong acidification.

Influence of dextran synthesized in situ on the rheological, technological and nutritional properties of whole grain pearl millet bread.

The effect of dextran produced in situ by Weissella confusa on the structure and nutrition quality of whole grain pearl millet bread containing 50% of wheat flour was investigated. NMR spectroscopy analysis indicated that the dextran formed by the strain consisted of a α-(1 → 6)-linked linear backbone and 3% α-(1 → 3) branches, and had a molar mass of 3.3 × 106 g/mol. In situ production resulted in 3.5% dextran (DW) which significantly enhanced the dough extensional properties, increased the bread specific volume (∼13%) and decreased crumb firmness (∼43%), moisture loss (∼15%) and staling rate (∼10%), compared to the control millet bread. DSC analysis showed that amylopectin recrystallization was significantly reduced in the bread containing dextran. In situ dextran production altered the nutritional value of millet, leading to increased free phenolic content (∼30%) and antioxidant activity. It also markedly lowered the bread predicted glycemic index and improved in vitro protein digestibility.

NMR spectroscopic analysis of exopolysaccharides produced by Leuconostoc citreum and Weissella confusa.

Dextrans are the main exopolysaccharides produced by Leuconostoc species. Other dextran-producing lactic acid bacteria include Streptococci, Lactobacilli, and Weissella species. Commercial production and structural analysis has focused mainly on dextrans from Leuconostoc species, particularly on Leuconostoc mesenteroides strains. In this study, we used NMR spectroscopy techniques to analyze the structures of dextrans produced by Leuconostoc citreum E497 and Weissella confusa E392. The dextrans were compared to that of L. mesenteroides B512F produced under the same conditions. Generally, W. confusa E392 showed better growth and produced more EPS than did L. citreum E497 and L. mesenteroides B512F. Both L. citreum E497 and W. confusa E392 produced a class 1 dextran. Dextran from L. citreum E497 contained about 11% alpha-(1-->2) and about 3.5% alpha-(1-->3)-linked branches whereas dextran from W. confusa E392 was linear with only a few (2.7%) alpha-(1-->3)-linked branches. Dextran from W. confusa E392 was found to be more linear than that of L. mesenteroides B512F, which, according to the present study, contained about 4.1% alpha-(1-->3)-linked branches. Functionality, whether physiological or technological, depends on the structure of the polysaccharide. Dextran from L. citreum E497 may be useful as a source of prebiotic gluco-oligosaccharides with alpha-(1-->2)-linked branches, whereas W. confusa E392 could be a suitable alternative to widely used L. mesenteroides B512F in the production of linear dextran.

Interactions between fava bean protein and dextrans produced by Leuconostoc pseudomesenteroides DSM 20193 and Weissella cibaria Sj 1b.

The aim of this study was to study the interactions between dextran and fava bean protein. Two dextrans produced by Leuconostoc pseudomesenteroides DSM 20193 and Weissella cibaria Sj 1b were purified and mixed with fava bean protein isolate (FPI) in water or in different buffers. The two isolated dextrans presented a typical dextran structure, mainly α-(1 → 6) linkages (above 95%) and few α-(1 → 3) branches, but they differed in molar mass and conformation. Dry-heating incubation of FPI and dextran mixture facilitated the conjugation of dextran to FPI through the Maillard reaction. Both mixed and conjugated systems were further heat-treated, and different influences of the formed covalent bonds on rheological properties were observed. The W. cibaria Sj 1b dextran had a much higher gel-strengthening ability than the Ln. pseudomesenteroides DSM 20193 dextran. The intermolecular FPI-dextran interactions played an important role in stabilizing the mixed systems at different pH.

The protective role of phytate in the oxidative degradation of cereal beta-glucans.

This study investigated the role of phytate in the Fenton reaction induced oxidative degradation of cereal ß-glucan. Viscosity analysis showed that the degradation rate was high in the beginning of oxidation, which fitted to the second order kinetic model. Oat ß-glucan contained significant amount of residual phytate and after the residual phytate was removed, faster degradation was shown compared to the original oat ß-glucan. Adding the same amount of phytic acid (PA) to the phytate removed ß-glucan sample also retarded the degradation but not as efficiently as the residual phytate. Considerable retardation of viscosity loss was shown when the PA to iron ratio was high. The presence of ascorbic acid weakened the retardation effect of phytic acid. Thus, phytate can significantly improve the oxidative stability of ß-glucan when the ratio of phytic acid to transition metals and the presence of ascorbic acid are taken into consideration.

Lipid oxidation induced oxidative degradation of cereal beta-glucan.

In food systems, lipid oxidation can cause oxidation of other molecules. This research for the first time investigated oxidative degradation of ß-glucan induced by lipid oxidation using an oil-in-water emulsion system which simulated a multi-phased aqueous food system containing oil and ß-glucan. Lipid oxidation was monitored using peroxide value and hexanal production while ß-glucan degradation was evaluated by viscosity and molecular weight measurements. The study showed that while lipid oxidation proceeded, ß-glucan degradation occurred. Emulsions containing ß-glucan, oil and ferrous ion showed significant viscosity and molecular weight decrease after 1 week of oxidation at room temperature. Elevated temperature (40°C) enhanced the oxidation reactions causing higher viscosity drop. In addition, the presence of ß-glucan appeared to retard the hexanal production in lipid oxidation. The study revealed that lipid oxidation may induce the degradation of ß-glucan in aqueous food systems where ß-glucan and lipids co-exist.

Lactose- and cellobiose-derived branched trisaccharides and a sucrose-containing trisaccharide produced by acceptor reactions of Weissella confusa dextransucrase.

Dextran-producing Weissella have received significant attention. However, except for maltose, the acceptor reactions of Weissella dextransucrases with different sugars have not been investigated. The action of recombinant Weissella confusa VTT E-90392 dextransucrase was tested with several potential acceptors, particularly, analogs lactose and cellobiose. The major acceptor products of both disaccharides were identified as branched trisaccharides, with a glucosyl residue α-(1 → 2)-linked to the acceptor's reducing end. An additional product, isomelezitose (6(Fru)-α-Glcp-sucrose), was also produced when using lactose as an acceptor. This is the first report of the synthesis of isomelezitose by a dextransucrase. The NMR spectra of the three trisaccharides were fully assigned, and their structures were confirmed by selective enzymatic hydrolysis. The trisaccharides prepared from (13)C6(glc) sucrose and lactose were analyzed by ESI-MS(n), and the fragmentation patterns of these compounds were characterized.

The oxidative degradation of barley ß-glucan in the presence of ascorbic acid or hydrogen peroxide.

Cereal ß-glucans are polysaccharides with health benefits that have been linked to their ability to increase luminal viscosity. However, the functional properties of cereal ß-glucans may be diminished by the susceptibility of this polysaccharide to oxidative degradation. In this study, barley ß-glucan was oxidised with hydrogen peroxide or ascorbic acid and the oxidative degradation of ß-glucan was investigated using both asymmetrical flow field-flow fractionation (AsFlFFF) with aqueous solvent and high performance size exclusion chromatography (HPSEC) with LiBr in DMSO as the solvent. Oxidation was shown to cause degradation of ß-glucan, the reaction being faster when oxidised with hydrogen peroxide compared with ascorbic acid. Both HPSEC and AsFlFFF showed comparable results as long as aggregates (only observed in AsFlFFF) were not included in the integration. The compact aggregates observed in oxidised samples suggest oxidation driven interactions between ß-glucan molecules.

Cloning and characterization of a Weissella confusa dextransucrase and its application in high fibre baking.

Wheat bran offers health benefits as a baking ingredient, but is detrimental to bread textural quality. Dextran production by microbial fermentation improves sourdough bread volume and freshness, but extensive acid production during fermentation may negate this effect. Enzymatic production of dextran in wheat bran was tested to determine if dextran-containing bran could be used in baking without disrupting bread texture. The Weissella confusa VTT E-90392 dextransucrase gene was sequenced and His-tagged dextransucrase Wc392-rDSR was produced in Lactococcus lactis. Purified enzyme was characterized using (14)C-sucrose radioisotope and reducing value-based assays, the former yielding K(m) and V(max) values of 14.7 mM and 8.2 µmol/(mg ∙ min), respectively, at the pH optimum of 5.4. The structure and size of in vitro dextran product was similar to dextran produced in vivo. Dextran (8.1% dry weight) was produced in wheat bran in 6 h using Wc392-rDSR. Bran with and without dextran was used in wheat baking at 20% supplementation level. Dextran presence improved bread softness and neutralized bran-induced volume loss, clearly demonstrating the potential of using dextransucrases in bran bioprocessing for use in baking.

Challenges in analysis of high-molar mass dextrans: comparison of HPSEC, AsFlFFF and DOSY NMR spectroscopy.

Dilute solutions of various dextran standards, a high-molar mass (HMM) commercial dextran from Leuconostoc spp., and HMM dextrans isolated from Weissella confusa and Leuconostoc citreum were analyzed with high-performance size-exclusion chromatography (HPSEC), asymmetric flow field-flow fractionation (AsFlFFF), and diffusion-ordered NMR spectroscopy (DOSY). HPSEC analyses were performed in aqueous and dimethyl sulfoxide (DMSO) solutions, while only aqueous solutions were utilized in AsFlFFF and DOSY. The study showed that all methods were applicable to dextran analysis, but differences between the aqueous and DMSO-based solutions were obtained for HMM samples. These differences were attributed to the presence of aggregates in aqueous solution that were less prevalent in DMSO. The study showed that DOSY provides an estimate of the size of HMM dextrans, though calibration standards may be required for each experimental set-up. To our knowledge, this is the first study utilizing these three methods in analyzing HMM dextrans.