Characteristics of Oat and Buckwheat Malt Grains for Use in the Production of Fermented Foods.

Malted gluten-free cereal grains and pseudo-cereals are interesting raw materials for producing fermented foods. The aim of the work was to assess selected technological quality characteristics and antioxidant properties of special malts in terms of use in the production of fermented foods. The research material consisted of malts made from oat, buckwheat, and brewing barley. Malting was performed on a microtechnical scale according to the standard scheme for brewing barley grain. The basic quality parameters of cereal grains obtained malts, and laboratory wort were assessed according to methods applicable in brewing. Atypical brewing malts were characterized by parameters such as malt extractability, protein solubilization, diastatic force, mash filtration time, and wort viscosity. The best results, comparable to barley malt, were obtained for naked oat malt. Malted buckwheat grains turned out to be the least biochemically modified, although their use in the production of beer and/or other fermented beverages is supported by the high content of bioactive substances and antioxidant potential. As the malting process of cereal plants improves their antioxidant properties and increases their nutritional value, oat and buckwheat malts can be successfully used to produce gluten-free fermented beverages or as an addition to fermented products, e.g., in baking and confectionery.

Influence of Pre-Treatment and Drying Methods on the Quality of Dried Carrot Properties as Snacks.

The aim of the current research was to evaluate the effect of pre-treatment and drying methods on the properties of dried carrots. Carrots were blanched (B) (1 or 3 min) or osmotic dehydrated (OD) (15 or 30 min) and dried by either convection (CD), microwave-convection (MW-CD), microwave-vacuum (MVD), or freeze-drying (FD). FD carrots showed the highest dry matter content (93.6-95.8%) and the lowest water activity (0.24-0.38). MVD carrots had lower dry matter content (79.5-95.8%) and two times more water activity (0.447-0.637) than FD. The highest color difference (∆E) in relation to raw material was noted in MVD samples (22-35) and the smallest in CD and FD (7-18), mainly due to the increase in brightness of the dried carrot. In general dried MCD carrot samples were characterized by the highest max force (hardness) (21.6-42.5 N; on average 34.7 N) in the breaking test and the lowest hardness was observed in the CD (10.8 N) ones. Pre-treatment and drying caused a significant decrease in the content of carotenoids (2.0-2.7 times) and chlorophyll (2.7-4.5 times) compared to the fresh carrot but a retention or increase in the total content of phenolics and antioxidant activity, especially in microwave-vacuum-dried carrots with an increase of even 2.7-2.9 times compared to raw material. High phenolic content (195.6-277.4 mg GA/100 g d.m.) was found in pre-osmotic dehydrated samples, and lower phenolic content was found in blanched samples (110.7-189.6 mg GA/100 g d.m.). Significantly, the highest average antioxidant activity was found in microwave-vacuum-dried samples (228.9 µmol Trolox/100 g d.m.). The results of this study indicate that microwave-vacuum-drying as an alternative to freeze-drying, including in combination with thermal or osmotic treatment, is very promising for the production of dried carrot snacks.

The Use of Brewer's Spent Grain after Beer Production for Energy Purposes.

The aim of this study was to assess the possibilities to use brewer's spent grains (BSGs) left over from beer production for energy purposes, and to determine its calorific value and chemical composition. The research materials were samples of wet spent grain from a brewery in Poland. Three samples, that are different in ingredient composition, were examined. The examined samples of BSGs were characterised by humidity that is typical for this product (approx. 77-80%). Convective drying of the spent grain contributed to a reduction in the water content in the biomass to below 10%. Samples of dry spent grain that were examined contained a similar amount of ash (3.8-4.1% d.m.) and organic matter (91.0-91.9% d.m.). All the examined spent grain samples demonstrated similar volatile matter content-approx. 77.8-78.7% d.m. and calorific value-approx. 15.6-15.9 MJ/kg. The estimated calorific value for wet samples (approx. 1.4-2.0 MJ/kg) indicated that it is necessary to lower water content in the biomass in order to improve its energy properties.

Dynamics of dark fermentation microbial communities in the light of lactate and butyrate production.

BACKGROUND: This study focuses on the processes occurring during the acidogenic step of anaerobic digestion, especially resulting from nutritional interactions between dark fermentation (DF) bacteria and lactic acid bacteria (LAB). Previously, we have confirmed that DF microbial communities (MCs) that fed on molasses are able to convert lactate and acetate to butyrate. The aims of the study were to recognize the biodiversity of DF-MCs able and unable to convert lactate and acetate to butyrate and to define the conditions for the transformation. RESULTS: MCs sampled from a DF bioreactor were grown anaerobically in mesophilic conditions on different media containing molasses or sucrose and/or lactate and acetate in five independent static batch experiments. The taxonomic composition (based on 16S_rRNA profiling) of each experimental MC was analysed in reference to its metabolites and pH of the digestive liquids. In the samples where the fermented media contained carbohydrates, the two main tendencies were observed: (i) a low pH (pH ≤ 4), lactate and ethanol as the main fermentation products, MCs dominated with Lactobacillus, Bifidobacterium, Leuconostoc and Fructobacillus was characterized by low biodiversity; (ii) pH in the range 5.0-6.0, butyrate dominated among the fermentation products, the MCs composed mainly of Clostridium (especially Clostridium_sensu_stricto_12), Lactobacillus, Bifidobacterium and Prevotella. The biodiversity increased with the ability to convert acetate and lactate to butyrate. The MC processing exclusively lactate and acetate showed the highest biodiversity and was dominated by Clostridium (especially Clostridium_sensu_stricto_12). LAB were reduced; other genera such as Terrisporobacter, Lachnoclostridium, Paraclostridium or Sutterella were found. Butyrate was the main metabolite and pH was 7. Shotgun metagenomic analysis of the selected butyrate-producing MCs independently on the substrate revealed C.tyrobutyricum as the dominant Clostridium species. Functional analysis confirmed the presence of genes encoding key enzymes of the fermentation routes. CONCLUSIONS: Batch tests revealed the dynamics of metabolic activity and composition of DF-MCs dependent on fermentation conditions. The balance between LAB and the butyrate producers and the pH values were shown to be the most relevant for the process of lactate and acetate conversion to butyrate. To close the knowledge gaps is to find signalling factors responsible for the metabolic shift of the DF-MCs towards lactate fermentation. Video Abstract.

Development of a High-Fibre Multigrain Bar Technology with the Addition of Curly Kale.

The aim of this study was to find the effect of kale and dietary fibre (DF) on the physicochemical properties, nutritional value and sensory quality of multigrain bars. A recipe of multigrain bars was prepared with the addition of fresh kale (20% and 30%) and DF preparations (apple, blackcurrant, chokeberry and hibiscus). The bars were baked at 180 °C for 20 min. These snack bars, based on pumpkin seeds, sunflower seeds, flaxseed and wholegrain oatmeal, are a high-calorie product (302-367 kcal/100 g). However, the composition of the bars encourages consumption. In addition to the ability to quickly satisfy hunger, such bars are rich in many natural ingredients that are considered pro-health (high fibre content (9.1-11.6 g/100 g), protein (11.2-14.3 g/100 g), fat (17.0-21.1 g/100 g, including unsaturated fatty acids), carbohydrates (20.5-24.0 g/100 g), as well as vitamins, minerals and a large number of substances from the antioxidant group. The addition of kale caused a significant increase of water content, but reduction in the value of all texture parameters (TPA profiles) as well as calorific values. The content of polyphenols was strongly and positively correlated with the antioxidant activity (r = 0.92). In the bars with 30% addition of kale (422 mg GA/100 g d.m.), the content of polyphenols was significantly higher than based ones (334 mg GA/100 g d.m.). Bars with the addition of the DF were characterized by a higher antioxidant activity, and the content of carotenoids, chlorophyll A and B and polyphenols. High sensory quality was demonstrated for all (from 4.8 to 7.1 on a 10-point scale). The addition of fibre preparations was also related to technological aspects and allows to create attractive bars without additional chemicals.

Evaluation of acidogenesis products' effect on biogas production performed with metagenomics and isotopic approaches.

BACKGROUND: During the acetogenic step of anaerobic digestion, the products of acidogenesis are oxidized to substrates for methanogenesis: hydrogen, carbon dioxide and acetate. Acetogenesis and methanogenesis are highly interconnected processes due to the syntrophic associations between acetogenic bacteria and hydrogenotrophic methanogens, allowing the whole process to become thermodynamically favorable. The aim of this study is to determine the influence of the dominant acidic products on the metabolic pathways of methane formation and to find a core microbiome and substrate-specific species in a mixed biogas-producing system. RESULTS: Four methane-producing microbial communities were fed with artificial media having one dominant component, respectively, lactate, butyrate, propionate and acetate, for 896 days in 3.5-L Up-flow Anaerobic Sludge Blanket (UASB) bioreactors. All the microbial communities showed moderately different methane production and utilization of the substrates. Analyses of stable carbon isotope composition of the fermentation gas and the substrates showed differences in average values of δ13C(CH4) and δ13C(CO2) revealing that acetate and lactate strongly favored the acetotrophic pathway, while butyrate and propionate favored the hydrogenotrophic pathway of methane formation. Genome-centric metagenomic analysis recovered 234 Metagenome Assembled Genomes (MAGs), including 31 archaeal and 203 bacterial species, mostly unknown and uncultivable. MAGs accounted for 54%-67% of the entire microbial community (depending on the bioreactor) and evidenced that the microbiome is extremely complex in terms of the number of species. The core microbiome was composed of Methanothrix soehngenii (the most abundant), Methanoculleus sp., unknown Bacteroidales and Spirochaetaceae. Relative abundance analysis of all the samples revealed microbes having substrate preferences. Substrate-specific species were mostly unknown and not predominant in the microbial communities. CONCLUSIONS: In this experimental system, the dominant fermentation products subjected to methanogenesis moderately modified the final effect of bioreactor performance. At the molecular level, a different contribution of acetotrophic and hydrogenotrophic pathways for methane production, a very high level of new species recovered, and a moderate variability in microbial composition depending on substrate availability were evidenced. Propionate was not a factor ceasing methane production. All these findings are relevant because lactate, acetate, propionate and butyrate are the universal products of acidogenesis, regardless of feedstock.

Dynamics and Complexity of Dark Fermentation Microbial Communities Producing Hydrogen From Sugar Beet Molasses in Continuously Operating Packed Bed Reactors.

This study describes the dynamics and complexity of microbial communities producing hydrogen-rich fermentation gas from sugar-beet molasses in five packed-bed reactors (PBRs). The bioreactors constitute a part of a system producing hydrogen from the by-products of the sugar-beet industry that has been operating continuously in one of the Polish sugar factories. PBRs with different working volumes, packing materials, construction and inocula were tested. This study focused on analysis (based on 16S rRNA profiling and shotgun metagenomics sequencing) of the microbial communities selected in the PBRs under the conditions of high (>100 cm3/g COD of molasses) and low (<50 cm3/g COD of molasses) efficiencies of hydrogen production. The stability and efficiency of the hydrogen production are determined by the composition of dark fermentation microbial communities. The most striking difference between the tested samples is the ratio of hydrogen producers to lactic acid bacteria. The highest efficiency of hydrogen production (130-160 cm3/g COD of molasses) was achieved at the ratios of HPB to LAB ≈ 4:2.5 or 2.5:1 as determined by 16S rRNA sequencing or shotgun metagenomics sequencing, respectively. The most abundant Clostridium species were C. pasteurianum and C. tyrobutyricum. A multiple predominance of LAB over HPB (3:1-4:1) or clostridia over LAB (5:1-60:1) results in decreased hydrogen production. Inhibition of hydrogen production was illustrated by overproduction of short chain fatty acids and ethanol. Furthermore, concentration of ethanol might be a relevant marker or factor promoting a metabolic shift in the DF bioreactors processing carbohydrates from hydrogen-yielding toward lactic acid fermentation or solventogenic pathways. The novelty of this study is identifying a community balance between hydrogen producers and lactic acid bacteria for stable hydrogen producing systems. The balance stems from long-term selection of hydrogen-producing microbial community, operating conditions such as bioreactor construction, packing material, hydraulic retention time and substrate concentration. This finding is confirmed by additional analysis of the proportions between HPB and LAB in dark fermentation bioreactors from other studies. The results contribute to the advance of knowledge in the area of relationships and nutritional interactions especially the cross-feeding of lactate between bacteria in dark fermentation microbial communities.

Cell factories converting lactate and acetate to butyrate: Clostridium butyricum and microbial communities from dark fermentation bioreactors.

BACKGROUND: Interactions between microorganisms during specific steps of anaerobic digestion determine metabolic pathways in bioreactors and consequently the efficiency of fermentation processes. This study focuses on conversion of lactate and acetate to butyrate by bacteria of dark fermentation. The recently recognized flavin-based electron bifurcation as a mode of energy coupling by anaerobes increases our knowledge of anaerobic lactate oxidation and butyrate formation. RESULTS: Microbial communities from dark fermentation bioreactors or pure culture of Clostridium butyricum are able to convert lactate and acetate to butyrate in batch experiments. The ability of C. butyricum to transform lactate and acetate to butyrate was shown for the first time, with ethanol identified as an additional end product of this process. A search for genes encoding EtfAB complexes and their gene neighbourhood in C. butyricum and other bacteria capable of lactate and acetate conversion to butyrate as well as butyrate-producers only and the lactate oxidiser Acetobacterium woodii, revealed that the Etf complexes involved in (i) lactate oxidation and (ii) butyrate synthesis, form separate clusters. There is a more extent similarity between Etf subunits that are involved in lactate oxidation in various species (e.g. A. woodii and C. butyricum) than between the different etf gene products within the same species of butyrate producers. A scheme for the metabolic pathway of lactate and acetate transformation to butyrate in C. butyricum was constructed. CONCLUSIONS: Studies on the conversion of lactate and acetate to butyrate by microbial communities from dark fermentation bioreactors or Clostridium butyricum suggest that a phenomenon analogous to cross-feeding of lactate in gastrointestinal tract also occurs in hydrogen-yielding reactors. A scheme of lactate and acetate transformation pathway is proposed, based on the example of C. butyricum, which employs flavin-based electron bifurcation. This process utilizes electron-transferring flavoprotein (Etf) complexes specific for (i) lactate oxidation and (ii) butyrate formation. Phylogenetic analysis revealed that such complexes are encoded in the genomes of other bacteria capable of lactate and acetate conversion to butyrate. These findings contribute significantly to our understanding of the metabolic pathways and symbiotic interactions between bacteria during the acidogenic step of anaerobic digestion.

Noteworthy Facts about a Methane-Producing Microbial Community Processing Acidic Effluent from Sugar Beet Molasses Fermentation.

Anaerobic digestion is a complex process involving hydrolysis, acidogenesis, acetogenesis and methanogenesis. The separation of the hydrogen-yielding (dark fermentation) and methane-yielding steps under controlled conditions permits the production of hydrogen and methane from biomass. The characterization of microbial communities developed in bioreactors is crucial for the understanding and optimization of fermentation processes. Previously we developed an effective system for hydrogen production based on long-term continuous microbial cultures grown on sugar beet molasses. Here, the acidic effluent from molasses fermentation was used as the substrate for methanogenesis in an upflow anaerobic sludge blanket bioreactor. This study focused on the molecular analysis of the methane-yielding community processing the non-gaseous products of molasses fermentation. The substrate for methanogenesis produces conditions that favor the hydrogenotrophic pathway of methane synthesis. Methane production results from syntrophic metabolism whose key process is hydrogen transfer between bacteria and methanogenic Archaea. High-throughput 454 pyrosequencing of total DNA isolated from the methanogenic microbial community and bioinformatic sequence analysis revealed that the domain Bacteria was dominated by Firmicutes (mainly Clostridia), Bacteroidetes, δ- and γ-Proteobacteria, Cloacimonetes and Spirochaetes. In the domain Archaea, the order Methanomicrobiales was predominant, with Methanoculleus as the most abundant genus. The second and third most abundant members of the Archaeal community were representatives of the Methanomassiliicoccales and the Methanosarcinales. Analysis of the methanogenic sludge by scanning electron microscopy with Energy Dispersive X-ray Spectroscopy and X-ray diffraction showed that it was composed of small highly heterogeneous mineral-rich granules. Mineral components of methanogenic granules probably modulate syntrophic metabolism and methanogenic pathways. A rough functional analysis from shotgun data of the metagenome demonstrated that our knowledge of methanogenesis is poor and/or the enzymes responsible for methane production are highly effective, since despite reasonably good sequencing coverage, the details of the functional potential of the microbial community appeared to be incomplete.

[The evaluation of anti-nutritive components in beer on the example of oxalic acid]. / Ocena zawartosci skladników antyodzywczych w piwie na przykladzie kwasu szczawiowego.

BACKGROUND: Food in its composition contains anti-nutritional substances that reduces or prevents the use of valuable nutrients. The oxalic acid, as phytate and dietary fiber, occurs naturally in foods of plant origin, to which the beer is classified. The negative effect of oxalic acid is reducing the bioavailability of calcium and magnesium, and disorder of metabolism of the body's absorption of these elements from the diet. The excess of oxalic acid and its salt in the diet contributes to the formation of certain diseases, such as oxalate urolithiasis, osteoporosis, arthritis, etc. Due to the diuretic effect of beer, drinking moderate amounts of it is recommended as a preventive and support urinary tract disorders. OBJECTIVE: The aim of this study was to determine and comparison the oxalic acid content in selected beers available on the Polish market. MATERIAL AND METHOD: Fifty seven samples of beer were used for this study. These samples were divided into three groups depending on the alcohol concentration declared by the producers (1st group--below 5.5% vol., 2nd group--from 5.5 to 6.5% vol., 3rd group--above 6.5% vol.). The beer samples were incubated in the ultrasonic bath for 15 minutes following pH adjustments up to pH = 2 with the 1 mol/L hydrochloric acid to transform calcium oxalates into soluble form, then filtered. The oxalic acid concentration was measured by high performance liquid chromatography (HPLC) with conductivity detection. RESULTS: The concentration of oxalic acid in tested samples of beer ranged from 1.8 to 30.3 mg/L. No considerable differences between the concentration of oxalic acid in the three tested group of beer with the various content of the alcohol were found. Basing on the average concentrations of the oxalic acid in the different groups of the tested beers the positive trend in oxalic acid concentration related to the increase of alcohol could be observed. CONCLUSIONS: The very low concentration of oxalic acid allows to classify beer as food product safe for the human health in terms of low oxalates content.