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. 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. 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.
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.
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 purpose of this study was to evaluate the possibility of using polyols (erythritol, xylitol, and maltitol) as alternatives to sucrose commonly used for osmotic dehydration (OD) of apples and to determine their content in osmotically dehydrated and dried fruit. For comparison, apples were also dehydrated in a sucrose solution. The content of sugars (sucrose, fructose, and glucose) was determined as well. Osmotic dehydration was examined at a temperature of 40 °C in time ranges between 15 and 360 min, and also 1440 min. The polyol solutions were equally or significantly more effective in the OD of apples compared to the sucrose solution. Regardless of the type of osmotic solution, the water loss of apples was from 2 to 10 times greater than solid gain. The color of fruit dehydrated in sucrose and xylitol solutions was characterized by the smallest total color difference (7.9–10.5) compared with the fresh apple tissue. Osmotic dehydration of apples in polyol solutions resulted in the transfer of these substances to fruit tissue. Erythritol was the most effective osmotic agent. The use of polyols for the OD of fruit at the pretreatment stage and the appropriate drying method can be used to create products with high qualities among others due to the lack of added sugars. The use of hybrid drying after the osmotic treatment is a good alternative to the freeze-drying process.
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.
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.