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Due to a large adaptability to different cultivation conditions and limited input compared to other cereals, sorghum is considered an emerging crop. Its antioxidant properties, high fiber content and low glycemic index also make it a valuable addition to a healthy diet, nevertheless, the presence of antinutritional factors and the lack of gluten, hamper its use as food ingredient. This study investigated the impact of sourdough fermentation on sorghum nutritional quality. Lactic acid bacteria dominating sorghum flour and sourdough were identified by culture-dependent analysis revealing Lactiplantibacillus plantarum as the dominant species found in the mature sourdough, whereas Weissella cibaria and Weissella paramesenteroides were the species isolated the most after the first refreshment. Among yeasts, Saccharomyces cerevisiae was the most prevalent. Lactic acid bacteria pro-technological and functional performances as starter were evaluated in sorghum type-II sourdoughs through an integrated characterization combining chromatographic and NMR spectroscopic techniques. The metabolic profile of the strains mainly grouped together W. cibaria strains and W. paramesenteroides AI7 which distinguished for the intense proteolysis but also for the presence of compounds particularly interesting from a physiological perspective (allantoin, glutathione, γ-aminobutyric acid and 2-hydroxy-3-methylbutyric acid), whose concentration increased during fermentation in a species or strain specific matter., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Carlo Giuseppe Rizzello reports financial support was provided by European Union Next-GenerationEU. If there are other authors, they declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)

Fungi are the main microorganisms responsible for the spoilage of bakery products, and their control and subsequent reduction of food waste are significant concerns in the agri-food industry. Synthetic preservatives are still the most used compounds to reduce bakery product spoilage. On the other hand, studies have shown that biopreservation can be an attractive approach to overcoming food and feed spoilage and increasing their shelf-life. However, limited studies show the preservation effects on real food matrices. Therefore, this study aimed to investigate the influence of microorganisms such as lactic acid bacteria (LAB) and yeasts on the growth of spoilage filamentous fungi (molds) on bread and panettones. In general, on conventional and multigrain bread, treatments containing Limosilactobacillus fermentum IAL 4541 and Wickerhamomyces anomalus IAL 4533 showed similar results when compared to the negative control (calcium propionate) in delaying the fungal growth of the tested species (Aspergillus chevalieri, Aspergillus montevidensis, and Penicillium roqueforti). Different from bread, treatments with W. anomallus in panettones delayed the A. chevalieri growth up to 30 days, 13 days longer than observed on negative control (without preservatives). This study showed that biopreservation is a promising method that can extend bakery products' shelf-life and be used as an alternative to synthetic preservatives., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Type 3 sourdoughs, which are starter culture-initiated and subsequently backslopped, are less studied than other sourdough types. Yet, they can serve as a model to assess how competitive starter culture strains for sourdough production are and how the microbial composition of such sourdoughs may evolve over time. In the present study, Limosilactobacillus fermentum IMDO 130101 was used to produce Type 3 sourdoughs, prepared from wheat and wholemeal wheat flours. Therefore, an initial fermentation of the flour-water mixture was performed at 30 °C for 48 h. This was followed by cold storage-backslopping cycles, consisting of refreshments (50 %, v/v), fermentation steps of 16 h, and storage at 4 °C each week, every three weeks, and every six weeks. The microbial dynamics (culture-dependent and -independent approaches) and metabolite dynamics were measured. In all sourdoughs produced, starter culture strain monitoring, following an amplicon sequence variant approach, showed that Liml. fermentum IMDO 130101 prevailed during one month when the sourdoughs were refreshed each week, during 24 weeks when the sourdoughs were refreshed every three weeks, and during 12 weeks when the sourdoughs were refreshed every six weeks. This suggested the competitiveness and robustness of Liml. fermentum IMDO 130101 for a considerable duration but also showed that the strain is prone to microbial interference. For instance, Levilactobacillus brevis and Pediococcus spp. prevailed upon further cold storage and backslopping. Also, although no yeasts were inoculated into the flour-water mixtures, Kazachstania unispora, Torulaspora delbrueckii, and Wickerhamomyces anomalus were the main yeast species found. They appeared after several weeks of storage and backslopping, which however indicated the importance of an interplay between LAB and yeast species in sourdoughs. The main differences among the mature sourdoughs obtained could be explained by the different flours used, the refreshment conditions applied, and the sampling time (before and after backslopping). Finally, the metabolite quantifications revealed continued metabolite production during the cold storage periods, which may impact the sourdough properties and those of the breads made thereof., Competing Interests: Declaration of competing interest The authors declare that no commercial or financial relationships were conflicting with the research, and that, as such, there is no conflict of interest., (Copyright © 2023 The Authors. Published by Elsevier B.V. All rights reserved.)

Thirty strains of lactic acid bacteria (LAB) and Saccharomyces cerevisiae E8.9 (wild type) were used to formulate fifteen combinations of starters by mixing two or three LAB with the yeast (ratio LAB: yeast, 10: 1). Such combinations were used to prepare rye sourdough and their performance in term of acidification and biochemical characteristics during fermentation at two temperatures (30 and 37 °C) and duration (4 and 8 h) were screened. The best thirteen sourdough formulations were selected and used for rye crispbread making. The analysis of acrylamide concentration demonstrated that 11 out 13 formulations resulted in significant decreases of concentration compared to the baker's yeast (control), with reductions up to 79.6 %. The rye sourdough crispbreads showed also higher amount of volatile organic compounds (VOCs) compared to the baker's yeast control. Two rye sourdough crispbreads, selected to represent the opposite extremes within the thirteen formulations in term of VOC profiles and fermentation performances, demonstrated better sensory and nutritional features, such as phytic acid reduction (up to 47.3 %), and enhanced total free amino acid compared to the control. These evidences suggest the potential of tailored sourdough fermentations as alternative and suitable biotechnological strategy for lowering acrylamide levels in rye crispbread., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Sourdough fermentation is attracting growing attention because of its positive effects on properties of leavened baked good. However, the changes in dough features and the mechanisms behind them are not well understood, which limits its widespread use. In this study, we assessed the effects of representative lactic acid bacteria in sourdough monoculture or co-culture with yeasts on dough characteristics. Physicochemical analysis identified increased proteolysis and enhanced nutritional properties of co-culture groups. However, a reduction in organic acids contents of co-culture groups compared to monoculture was detected, and this effect was not limited by the yeast species. The RNA sequencing further demonstrated that the presence of yeast enhanced the protein metabolic activity of lactic acid bacteria, while decreased its organic acid biosynthetic activity. Moreover, the proteomic analysis revealed that endogenous metabolic proteins of flour, such as pyruvate kinase, glucosyltransferase and pyruvate dehydrogenase play a key role in carbohydrate metabolism during fermentation. This study uncovered the influence of typical microorganisms and endogenous enzymes on dough characteristics based on different aspects. Bacteria-mediated consumption of proteins and increased proteolysis in co-culture groups may underlie the improved digestibility and nutritional effects of sourdough fermented products, which provides an important basis for nutrient fortified bread making with multi-strain leavening agent., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

The final quality of wholemeal wheat bread is determined by the process parameter settings and leavening strategy. We hypothesise that the used leavening strategy may influence the optimal process parameter settings and, as such, the specific volume of the bread loaf. To analyse this interaction, bread was leavened with (i) a type 1 sourdough (SB), (ii) a type 1 sourdough combined with baker's yeast (YSB), or (iii) baker's yeast (YB). For each leavening strategy, the specific volume of bread, in response to variations in mixing time (4-10/4-14 min), water absorption (60-85 %), and proofing time (1-7/1-3 h), was analysed using an I-optimal response surface experimental design. Data modelling identified a substantially lower maximal specific volume of SB (2.13 mL/g), compared to YSB (3.30 mL/g) and YB (3.26 mL/g). The proofing time and water absorption mostly influenced the specific volume of the SB and YSB, respectively. However, the mixing and proofing times mainly affected the specific volume of YB. The type 1 sourdough reduced the mixing time and water absorption required for an optimal specific volume of bread compared to baker's yeast. These results challenge the idea of yielding higher volumes upon using sourdough compared to baker's yeast and highlight the importance of optimisation of bread dough formulations and breadmaking processes., Competing Interests: Conflict of interest The authors declare that no commercial or financial relationships were conflicting with the research, and that, as such, there is no conflict of interest., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Almond production generates large amounts of by-products rich in polyphenols. In this study, almond skin was explored as a valuable food ingredient in bread making. To this purpose, almond skin was used to produce functional products modifying a traditional sourdough bread recipe. The doughs were prepared replacing semolina with powdered almond skin (PAS) at 5 and 10 % (w/w). Sourdough inoculum was started with a mix of lactic acid bacteria (LAB) and propagated in semolina until reaching pH 3.7. The pH of PAS added breads was higher than that of control (CTR) breads before and after fermentation. Plate counts showed a similar evolution of LAB and total mesophilic microorganisms, but members of Enterobacteriaceae and coliform were detectable in PAS doughs. Illumina data clearly showed a dominance of lactobacilli in all trials, but PAS doughs displayed the presence of Bacillus. The final bread characteristics were influenced by PAS and its addition percentage; in particular, crust and crumb colour resulted darker, the alveolation decreased and, regarding sensory attributes, odour intensity increased, while bread odour diminished. In presence of PAS, bread emissions were characterized by lower percentages of alcohols and aromatic hydrocarbons and higher percentages of the other volatile compound classes, especially terpenoids like β-pinene, β-myrcene and limonene than CTR trial. After in vitro simulated digestion, the final release of phytochemicals from 10 % PAS bread was almost 100 %. Thus, PAS determined an increase of the antioxidant capacity of the breads. Phytochemicals released from digested PAS-fortified bread can provide antioxidant protection in a complex biological environment such as human intestinal-like cells. Besides the positive functional properties of PAS, this work also evidenced the hygienic issues of almond skin and, in order to avoid potential risks for the human health, highlighted the need to preserve its microbiological characteristics during storage for their reuse in bread production., Competing Interests: Declaration of competing interest The authors declare that there is no conflict of interest for this research., (Copyright © 2023 The Authors. Published by Elsevier B.V. All rights reserved.)

In plants, hydroxycinnamic and hydroxybenzoic acids occur mainly as esters. This study aimed to determine the contribution of individual phenolic acid esterases in Lp. plantarum TMW1.460, which encodes for four esterases: TanA, Lp_0796, Est_1092 and a homolog of Lj0536 and Lj1228 that was termed HceP. To determine which of the phenolic acid esterases present in Lp plantarum TMW1.460 are responsible for esterase activity, mutants with deletions in lp_0796, est_1092, tanB, hceP, or hceP and est_1092 were constructed. The phenotype of wild type strain and mutants was determined with esters of hydroxycinnamic acids (chlorogenic acid and ethyl ferulate) and of hydroxybenzoic acids (methyl gallate, tannic acid and epigallocatechin-3-gallate). Lp. plantarum TMW1.460 hydrolysed chlorogenic acid, methyl ferulate and methyl gallate but not tannic acid or epigallocatechin gallate. The phenotype of mutant strains during growth in mMRS differed from the wild type as follows: Lp. plantarum TMW1.460ΔhceP did not hydrolyse esters of hydroxycinnamic acids; Lp. plantarum TMW1.460ΔtanB did not hydrolyse esters of hydroxybenzoic acids; disruption of est_1092 or Lp_0796 did not alter the phenotype. The phenotype of Lp. plantarum TMW1.460ΔΔhceP/est_1092 was identical to Lp. plantarum TMW1.460ΔhceP. The metabolism of phenolic acids during growth of the mutant strains in broccoli puree and wheat sourdough did not differ from metabolism of the wild type strain. In conclusion, esters of hydroxycinnamic and hydroxybenzoic acids each are hydrolysed by dedicated enzymes. The hydroxycinnamic acid esterase HceP is not expressed, or not active during growth of Lp. plantarum TMW1.460 in all food substrates., Competing Interests: Declaration of competing interest The authors declare no conflict of interest., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Previous researchers have shown the potential of sourdough and isolated lactic acid bacteria in reducing wheat allergens. As the interactions of lactic acid bacteria with yeast is a key event in sourdough fermentation, we wished to investigate how yeast affects metabolism of lactic acid bacteria, thereby affecting protein degradation and antigenic response. In this study, three strains isolated from sourdough were selected for dough fermentation, namely Pediococcus acidilactici XZ31, Saccharomyces cerevisiae JM1 and Torulaspora delbrueckii JM4. The changes in dough protein during the fermentation process were studied. Protein degradation and antigenic response in dough inoculated with Pediococcus acidilactici XZ31 monoculture and co-culture with yeast were mainly evaluated by SDS-PAGE, immunoblotting, ELISA and Liquid chromatography-tandem mass spectrometry assay. The whole-genome transcriptomic changes in Pediococcus acidilactici XZ31 were also investigated by RNA sequencing. The results showed that water/salt soluble protein and Tri a 28/19 allergens content significantly decreased after 24 h fermentation. Co-culture fermentation accelerated the degradation of protein, and reduced the allergen content to a greater extent. RNA-sequencing analysis further demonstrated that the presence of yeast could promote protein metabolism in Pediococcus acidilactici XZ31 for a certain period of time. These results revealed a synergistic effect between Pediococcus acidilactici XZ31 and yeast degrading wheat allergens, and suggested the potential use of the multi-strain leavening agent for producing hypoallergenic wheat products., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Knowledge of the effects of various strains of acetic acid bacteria (AAB) on sourdough remains limited. In this study, the diversity of microbial taxa in sourdoughs fermented by different starters was assessed and their functional capacity was evaluated via high-throughput metagenomics sequencing. Results showed that Erwinia (29.43%), Pantoea (45.89%), and Enterobacter (9.16%) were predominant in the blank CK treatment. Lactobacillus (91.40%), Saccharomyces (6.13%), as well as the AAB genus Acetobacter (0.61%) were the dominant microbial genera in the sourdoughs started by yeast and a strain of lactic acid bacteria (YL treatment). By contrast, the dominant genera in the sourdoughs started by yeasts and various LAB and AAB strains (YLA treatment) were Komagataeibacter (0.39%) except for the inoculated Lactobacillus (68.37%), Acetobacter (20.17%), and Saccharomyces (8.31%) species. Functional prediction of these changes in microbial community and diversity revealed that various metabolism-related pathways, including alanine, aspartate, and glutamate metabolism (21.95%), as well as amino acid biosynthesis (19.14%), were predominant in the sourdoughs started by yeast and an AAB strain (YA treatment). Moreover, arginine biosynthesis (11.65%) were the dominant pathways in the YL treatment. The fermented dough added with sourdoughs started with yeast + AAB and yeast + AAB + LAB strains had substantially higher contents (more than 48.58% in total) of essential amino acids than the dough added with sourdoughs started with yeast + LAB strain. These results demonstrated that amino acid biosynthesis has a beneficial effect on sourdoughs inoculated with an AAB strain., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Extra ingredients are often used in traditional sourdough production recipes by artisan bakeries. These ingredients may be the source of microorganisms or stimulate the growth and/or the metabolic activities of the microorganisms added to or naturally present in the flour-water mixture. The present study examined the influence of the addition of lemon juice or apple juice as source of citrate or malate, respectively, on the growth and activity of the citrate- and malate-positive Companilactobacillus crustorum LMG 23699 strain (formerly known as Lactobacillus crustorum LMG 23699), used to initiate firm (dough yield of 200) wheat sourdough productions, and on the flavour of the baked goods produced. Three fermentation strategies were applied, namely one-step long fermentation sourdough production processes with the addition of juice at the start (Type 2) and backslopped fermentations with the addition of juice either only at the start of the sourdough productions or at the start of the sourdough productions and at the beginning of each subsequent refreshment step during the whole backslopping process (both Type 3). It turned out that the starter culture strain used prevailed during all sourdough productions performed. Yeasts were particularly present in Type 3 sourdough productions, although lemon juice retarded their growth. Due to high yeast activity, high concentrations of ethanol and glycerol were produced toward the end of the sourdough productions. Addition of lemon juice stimulated the production of lactic acid, acetic acid, and the buttery flavour compounds acetoin and diacetyl, because of citrate conversion, during the Type 2 and Type 3 sourdough productions. In Type 3 sourdough productions, these compounds were found in higher concentrations only when lemon juice was added at each backslopping step. Alternatively, the addition of apple juice led to high concentrations of lactic acid because of malolactic fermentation in both Type 2 and Type 3 sourdough productions. Moreover, the addition of apple juice increased the initial concentrations of the carbohydrates (fructose, glucose, and sucrose) and sugar alcohols (mannitol and sorbitol), which were exhausted upon backslopping or accumulated in the sourdough matrix, respectively. Baked goods produced using sourdoughs obtained from the Type 2 and Type 3 sourdough productions with the addition of juice at each backslopping step were significantly different in flavour from doughs supplemented with the respective juices and lactic acid and/or Type 3 sourdough productions with the addition of juice only at the start., (Copyright © 2020 Elsevier B.V. All rights reserved.)

In the present work the microbial dynamics in wheat kernels were evaluated over time. The main aim of this research was to study the resistance of lactic acid bacteria (LAB) and yeasts associated to unprocessed cereals used for bread making during long term conservation. To this purpose four Triticum durum Desf. genotypes including two modern varieties (Claudio and Simeto) and two Sicilian wheat landraces (Russello and Timilia) were analysed by a combined culture-independent and -dependent microbiological approach after one, two or three years from cultivation and threshing. DNA based MiSeq Illumina technology was applied to reveal the entire bacterial composition of all semolina samples. The samples showed a different distribution of bacterial taxa per variety and time of storage. The groups mostly represented were Stenotrophomonas, Pseudomonas, Erwinia, Delftia and Sphingomonas genera, Enterobacteriaceae and Oxalobacteriaceae families, and Actinobacteria phylum. Among LAB, only Enterococcus genus was detected barely in a single sample (Simeto stored for one year) by the next generation sequencing, indicating that LAB remained unassigned or their abundances were below 0.1% or their DNAs were rendered inaccessible. Plate counts showed consistent differences in relation to genotypes and duration of storage, with the highest levels found for total mesophilic microorganisms detected up 6.8 Log CFU/g. Colonies of presumptive sourdough LAB were detected only in a few samples. Cocci constituted the major group of LAB in almost all samples. Following the enrichment procedure, almost all samples were characterised by the presence of acidifying microorganisms. All isolates collected before and after enrichment represented 28 different strains belonging to 10 species of Enterococcus, Lactobacillus and Pediococcus genera. The most resistant species during aging were Enterococcus faecium, Enterococcus durans, Lactobacillus brevis, Lactobacillus pentosus and Lactobacillus paracasei demonstrating that lactobacilli and enterococci are able to overcome the stressing conditions represented by cereal storage better than other LAB genera commonly found associated to cereals after harvest. Yeast community included mostly species with no interest in bread making., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Sourdough is a very interesting natural preservation system to prolong mould free shelf-life of bread. Numerous studies have reported that the antifungal activity of sourdough is mainly correlated with the presence of lactic (LA) and acetic acid (AA), but very few information is available on the effect of undissociated acid concentrations in the aqueous phase of bread (C HA ; mmole/L). This study was conducted to provide additional information about the mode of action of the acids in sourdough bread, enabling a better shelf-life prediction. This study was divided into two parts. In part 1, three industrial biological sourdoughs were characterized (dough yield, pH, a w , fermentation quotient, microbiota). During 7 weeks, a shelf-life test with natural flora was conducted with daily checks of visible mould growth (21 °C). In part 2, the effect of the acids present in the antifungal active sourdough breads was validated in chemically acidified wheat breads. Complete growth inhibition was observed in full-baked sourdough bread (30 g/100 g dough) containing Lactobacillus sanfranciscensis and Saccharomyces cerevisiae as dominant sourdough micro-organisms, whereas in control bread the shelf-life was limited to 4.4-9.2 days. These full-baked sourdough breads contained 36 mmole undissociated LA/L and 220 mmole undissociated AA/L. The data were used to make General Linear Regression models for shelf-life prediction and resulted in a fit of R 2  = 0.79 when expressing the shelf-life in function of C HA,LA and C HA,AA . In acidified breads, the role of lactic acid was not significant and only impacted shelf-life indirectly through acidification. No difference between antifungal activity of sourdough breads and chemically acidified bread with comparable C HA,AA concentrations was observed. Shelf-life increased when 150-200 mmole undissociated AA/L aqueous phase in bread was present. To conclude, this study showed the importance of the undissociated acid fraction of acetic acid in relation to bread shelf-life, together with bread pH and moisture content., (Copyright © 2020 Elsevier B.V. All rights reserved.)

The demand for sugar reduction in products across the food and beverage industries has evoked the development of novel processes including the application of fermentation with lactic acid bacteria. Heterofermentative lactic acid bacteria (LAB) are diverse in their ability to utilise fermentable sugars and can also convert fructose into the sweet tasting polyol, mannitol. The sourdough microbiota has long been recognised as an ecological niche for a range of homofermentative and heterofermentative lactic acid bacteria. A leading determinant in the biodiversity of sourdough microbial populations is the type of flour used. Ten non-wheat flours were used and back-slopped for 7 days resulting in the isolation of 52 mannitol producing isolates which spanned six heterofermentative species of the genera Lactobacillus, Leuconostoc and Weissella. Assessment of mannitol productivity in fructose concentrations up to 100 g/L found Leuconostoc citreum TR116, to have the best mannitol producing characteristics, consuming 95% of available fructose and yielding 0.68 g of mannitol per gram of fructose consumed which equates to the maximal theoretical yield. Investigation of the effects of initial pH on mannitol production and other fermentation parameters in the isolates found pH 7 to be best for isolates Lactobacillus brevis TR052, Leuconostoc fallax TR111, Leuconostoc citreum TR116, Leuconostoc mesenteroides TR154 and Weissella paramesenteroides TR212, while pH 6 was optimal for Leuconostoc pseudomesenteroides TR080. The fermentation of apple juice with each isolate resulted in sugar reduction ranging from 30.3-74.0 g/L (34-72%). When apple juice fermentation with Leuconostoc citreum TR116 was scaled up to 1 L bioreactor a reduction in sugar of 98.6 g/L (83%) was achieved along with the production of 61.6 g/L mannitol. This demonstrates a fermentative process for sugar reduction in fruit juice with concomitant production of the sweet metabolite mannitol to create a fermentate that is suitable for further development as a low sugar fruit juice alternative., Competing Interests: Declaration of competing interest The authors certify that they have NO affiliations with or involvement in any organization or entity with any financial interest (such as honoraria; educational grants; participation in speakers’ bureaus; membership, employment, consultancies, stock ownership, or other equity interest; and expert testimony or patent-licensing arrangements), or non-financial interest (such as personal or professional relationships, affiliations, knowledge or beliefs) in the subject matter or materials discussed in this manuscript., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Lactobacillus (L.) sanfranciscensis is a competitive key species in sourdough fermentations. However, the principles involved in establishing the commonly observed phenomenon of strain dominance are unresolved. This has been studied little because the methods for fast and reliable differentiation of strains and their monitoring during fermentation are tedious and cannot be done with large numbers of isolates. In this contribution, we present a strain-specific, PCR-based typing method that uses length heterogeneities of the clustered regularly interspaced short palindromic repeats (CRISPR) loci as they occur in the genomes of different strains. In silico analysis of 21 genomes revealed 14 different CRISPR genotypes. We then designed a primer set to simultaneously detect different strains in a multiplex PCR assay designated CRISPR locus length polymorphism PCR (CLLP-PCR). The usefulness of this method was evaluated in lab-scale sourdough fermentations conducted with rye and wheat flours. First, the flour was mixed with water to a dough yield of 200. Then each dough was inoculated with four different L. sanfranciscensis strains (TMW 1.1150, TMW 1.392, TMW 1.2142, and TMW 1.2138) at levels of 10 9  cfu/g each. Sourdoughs were propagated at 28 °C for 5 days by back slopping 5% to the flour mass every 24 h. Samples were collected each day; DNA was isolated, and the presence of strains was detected qualitatively in the sourdoughs with PCR. L. sanfranciscensis TMW 1.392 became dominant as early as 2 days into the fermentation and remained the only detectable strain for the rest of the sampling period. CLLP-PCR proved to be useful in investigating the assertiveness of different strains of L. sanfranciscensis in sourdoughs. Therefore, CLLP-PCR may be used as a tool to investigate assertiveness of microorganisms in food fermentations at the strain level., Competing Interests: Declaration of competing interest The authors declare no conflict of interest., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Chia, is a gluten-free, rich in proteins, oilseed that is "on trend" as an alternative ingredient in food production, adding nutritional value. As a reservoir of natural biodiversity, lactic acid bacteria development, during spontaneous chia flour fermentation (sourdough) for 10 days, were investigated by culturing and high throughput sequencing (HTS). Culture-dependent analysis showed a rapid increase in total LAB numbers from the second day of sourdough refreshment. Taxonomical identification of LAB isolates by rep-PCR and further 16S rRNA sequencing was performed. Besides Among identified LAB by culture-dependent approach, species from genus Enterococcus were the most abundant; Lactococcus (Lc. lactis), Lactobacillus (L. rhamnosus) and Weissella (W. cibaria) species were also isolated. By HTS, twelve OTUs belonging to LAB genera were identified during chia sourdough fermentation with an increased Lactobacillus diversity. Enterococcus (E.) faecium, E. mundtii, W. cibaria and L. rhamnosus were detected as dominant species in the final propagation stages while Bacillus and Clostridium were mostly present during first fermentation stages. The investigation of biotechnological and safety traits (acidification ability, protein hydrolysis, exopolysaccharides production, antimicrobial activity and antibiotic resistance) of 15 representative LAB strains was performed. Strains characterization led to the selection of Lc. lactis CH179, L. rhamnosus CH34 and W. cibaria CH28 as candidates to be used as novel functional starter culture for gluten-free chia fermented products. As far as we know, this is the first study providing information on the molecular inventory of LAB population during spontaneous fermentation of chia sourdough., Competing Interests: Declaration of competing interest No conflict of interest was declared., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Sourdough is used in production of (steamed) bread as leavening agent (type I sourdoughs) or as baking improver to enhance flavour, texture, and shelf life of bread (type II sourdoughs). The long-term propagation of sourdoughs eliminates dispersal limitation and consistently leads to sourdough microbiota that are composed of host adapted lactobacilli. In contrast, community assembly in spontaneous cereal fermentations is limited by dispersal and nomadic or environmental lactic acid bacteria are the first colonizers of these sourdoughs. Propagation of sourdoughs for use as sole leavening agent (type I sourdoughs) dictates fermentation conditions that select for rapid growth. Type I wheat- and rye sourdoughs are consistently populated by insect-adapted lactobacilli, particularly Lactobacillus sanfranciscensis, which is characterized by a small genome size and a restricted metabolic potential. The diverse fermentation conditions employed in industrial or artisanal Type II sourdough fermentation processes also result in a more diverse microbiota. Nevertheless, type II sourdoughs are typically populated by vertebrate host adapted lactobacilli of the L. delbrueckii and L. reuteri groups. Metabolic traits of host-adapted lactobacilli that enhance competitiveness in intestinal ecosystems also provide technological functionality in bread making. Examples include formation of exopolysaccharides, arginine-, glutamine- and glutamate based mechanisms of acid resistance, and glycosyl hydrolases that reduce FODMAP levels in sourdough and sourdough bread. In conclusion, consideration of the lifestyle of sourdough lactobacilli facilitates the selection of competitive and functional sourdough starter cultures., (Copyright © 2018 Elsevier B.V. All rights reserved.)

As one of the most traditional biotechnologies, sourdough fermentation has deep effects on rheology, sensory and shelf life attributes of baked goods. The most recent literature has also highlighted the effects of sourdough fermentations on several functional/nutritional features of baked goods. While some aspects such as the potential to lower glycemic index, increase mineral bioavailability and decrease the gluten content have been proven almost definitively, others potentialities are emerging, which deserve novel insights. This reviews reports the main evidence on the use of sourdough fermentation for salt reduction in baked goods, management of irritable bowel syndrome (IBS), synthesis/release of bioactive compounds, especially the metabolism of phenolic compounds, and exploitation of the potential of non-conventional flours (legumes and pseudo-cereals) and milling by-products (bran and germ). A brief description on the spiritual, cultural and functional/nutritional significance of leavened bread throughout centuries has also given., (Copyright © 2018 Elsevier B.V. All rights reserved.)

This study aimed to evaluate the effect of soft (Triticum aestivum) and durum (Triticum durum) wheat flours at different extraction rate (type 00, 0, 1 and 2) and wholemeal flours on the microbial composition and functionality of type I sourdough. Enterobacteriaceae constituted the main component of the microbiome of refined soft and durum wheat flours. On the contrary, wholemeal durum wheat flour harboured mainly Xanthomonadaceae. Differences were also found between the soft and durum wheat flours. After 8 h of fermentation (1 day), a different behaviour of the microbiome components was observed. All the mature sourdoughs harboured a core microbiome constituted by 4 species (Pediococcus pentosaceus, Lactobacillus brantae, Pediococcus argentinicus and Weissella cibaria). Based on the type of flour, the relative abundance of each core species differed among sourdoughs. In addition, other dominant lactic acid bacteria species were variously detected in sourdoughs. Mature sourdoughs showed marked variations for the concentrations of glucose, fructose, maltose, lactic and acetic acids, ethanol and free amino acids (FAA). Specific correlations (r > 0.7; FDR < 0.05) were found between the microbiome and total phenols, fibres and metabolome of mature sourdoughs. Breads made by using wholemeal soft wheat or durum wheat doughs started by wholemeal wheat flour-based sourdoughs were characterized by the highest total amount of free cations (Ca 2+ , Zn 2+ , Fe 2+ , Mg 2+ ), antioxidant activity and improved sensory traits., (Copyright © 2018. Published by Elsevier B.V.)

The application of omics methodologies helps to further unravel sourdough fermentation processes. Of all methodologies applied, metagenetics is the most used one to analyse sourdoughs, allowing to elucidate their microbial structure, albeit that it is based on the sequencing of a very small part of whole-community DNA. Although shotgun metagenomics, metatranscriptomics, metaproteomics, and meta-metabolomics are very promising to analyse sourdough fermentations and the terminology is often used in a confusing way, they have not been fully used sensu stricto yet. For instance, up to now, metatranscriptomics is restricted to the use of a functional gene microarray for lactic acid bacteria. Further, meta-metabolomics often deals with metabolite target analysis of sourdough fermentation samples to determine the actual concentrations of residual substrates and metabolites produced as well as to list their volatilome solely. In contrast, genomics has been applied several times, albeit that the whole-genome sequence of only one yeast strain and of 41 lactic acid bacterial strains, originally isolated from sourdoughs, is available. However, the genomics data accessible in public databases should be considered with caution because of inaccurate gene annotations, due to automated annotation pipelines, thereby possibly overruling original, high-quality, well-curated gene annotations., (Copyright © 2018 Elsevier B.V. All rights reserved.)

In recent years, recommendations on whole grains consumption have been added to the overall dietary guidelines of many countries around the world. Despite the many benefits on human health, whole grains contain several anti-nutritional factors which decrease their nutritional quality leading to a poor use in human diet. Here, an integrate biotechnological approach, combining germination and sourdough fermentation with selected lactic acid bacteria, was set-up in order to improve the functional and nutritional quality of wheat, barley, chickpea, lentil and quinoa grains. Sourdough fermentation with Lactobacillus rossiae LB5, Lactobacillus plantarum 1A7 and Lactobacillus sanfranciscensis DE9 further enhanced the nutritional and functional features of sprouted flours by means of increased peptides, free amino acids and γ‑aminobutyric acid concentrations, and decreased phytic acid, condensed tannins, raffinose and trypsin inhibitory activity. Sensory appreciable wheat breads fortified with the fermented sprouted flours were manufactured and characterized, showing high protein digestibility and low starch availability., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Previous investigations on pizza dough lactic acid bacteria (LAB) revealed that facultative heterofermentative species (FHS) were more represented than obligate heterofermentative species (OHS) within the Lactobacillus genus. Thus, the main hypothesis of this work was that facultative and obligate heterofermentative Lactobacillus species can impact differently the appreciation of baked pizza. The performances of different Lactobacillus, including L. sanfranciscensis, L. brevis and L. rossiae among OHS and L. plantarum, L. graminis and L. curvatus among FHS were tested in single or multiple combinations during pizza production. The values of pH, total titratable acidity and LAB levels indicated that the acidification process was almost comparable among trials. The fermentation quotient of FHS trials was above 4.0. All trials were dominated by the added LAB and for the trials with the multiple strain starter inoculums, the species found at the highest cell densities were L. sanfranciscensis, L. brevis and L. plantarum. Significant differences among pizzas were found for weight loss, colour, morphology and volatile organic compounds (VOCs). The last analysis revealed the presence of eight chemical classes with aldehydes, esters, alcohols and acids as major compounds and allowed the separation of the trials FHS and OHS. Sensory attributes were significantly different for judges and pizzas and the most relevant differences were found for crust colour, presence of bubbles, resistance to tearing, crispness and chewiness. The overall assessment reached the highest scores for the mixed culture of OHS and FHS together., (Copyright © 2019 Elsevier B.V. All rights reserved.)

In traditional sourdough fermentation recipes of artisan bakeries, often extra ingredients are added to the flour-water mixture. This may accentuate the aroma and taste of the baked products produced from such sourdoughs. This is made possible, for instance, by stimulating certain microbial activities during fermentation. This study examined the effects of the addition of citrate (a food-grade organic acid present in milk and lemons) on wheat sourdough fermentation in the presence of a citrate-positive, homofermentative, lactic acid bacterial starter culture strain, namely Lactobacillus crustorum LMG 23699. Both liquid and firm wheat sourdoughs were produced. The starter culture strain was able to steer all wheat sourdough fermentations performed, as it always prevailed due to its competitiveness, as shown through culture-dependent microbiological plating and culture-independent bacterial community profiling. Moreover, it possessed all enzyme-encoding genes (as unraveled through genome mining) necessary to convert citrate into desirable compounds such as lactic acid, acetic acid, succinate, acetoin, diacetyl, and 2,3-butanediol. Indeed, citrate addition to the wheat flour-water mixture had an impact on the sourdough fermentation dynamics and thus on the aroma profile of the liquid and firm sourdoughs produced and breads made thereof. A higher final pH, higher total titratable acidity values, and low yeast counts were found in wheat sourdoughs produced with citrate. In particular, the starter culture strain added converted the supplemented citrate into more l-lactic acid as well as acetoin and diacetyl (buttery aroma compounds), which was independent of the dough yield. The buttery aroma compounds were also accentuated in the concomitant breads produced. Further, organic acid production was stimulated in the sourdoughs, whereas increased pyrazine concentrations occurred in the breads. Consequently, citrate supplementation to wheat sourdoughs could be of interest to produce baked goods with enhanced buttery aroma compounds and notes., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Sourdough fermentation of cereal foods is an excellent source of obtaining peptides due to the ability of lactic acid bacteria to activate cereal proteases and produce strain-specific peptidases. With the aim of identifying the lactic acid bacterial strains potentially most effective in producing bioactive peptides, 131 lactobacilli isolates from Italian sourdoughs, used in baking technology, have been screened for proteolytic and peptidase activity. Of these, 23 strains were selected and singly inoculated in liquid sourdoughs from which a Low Molecular Weight fraction containing peptides was obtained. Evaluation of the antioxidant and anti-inflammatory activities of the extracts was performed on cultured cells (RAW 264.7 murine macrophage, murine H-end endothelium cells and Human intestinal Caco-2 cells) by assaying Reactive Oxygen Species (ROS) content, NFkB/IkB expression level and Interleukin-1β production. As a result, three lactobacilli strains showed a high antioxidant and anti-inflammatory ability enabling the development of model sourdoughs that will potentially increase the nutritional benefits of bread., (Copyright © 2018 Elsevier B.V. All rights reserved.)

The formation of HoPS and oligosaccharides in sourdough fermentation improves bread quality but is dependent on the expression of glycansucrases by lactic acid bacteria. Data on the expression of dextransucrases by Weissella spp., however, are limited. This study therefore aimed to assess dextansucrase expression in W. cibaria 10 M, focusing on the effect of temperature. The effect of temperature on growth, oligosaccharide and dextran synthesis by W. cibaria 10 M was determined and the expression and activity of cell-associated dextransucrase from W. cibaria 10 M were investigated. The oligosaccharides profiles were measured by thin layer chromatography and high performance anion exchange chromatography coupled to pulsed amperometric detection. Dextran formation was quantified by size exclusion chromatography. W. cibaria grew fastest at 30 °C but oligosaccharide formation was highest at 20 °C or less. Dextransucrase expression as measured by reverse transcription quantitative PCR, SDS-PAGE, and activity of cell-associated dextransucrase were maximal at 15 °C. Cold shift incubation, characterized by incubation at 30 °C to obtain biomass, followed by shift to 6 °C to induce dextransucrase expression, supported high dextransucrase activity in laboratory media. Cold shift fermentation of wheat and sorghum sourdoughs supplemented with 15 or 30% sucrose increased the yields of oligosaccharides, and resulted in formation of 16 and 12 g/kg dextran in wheat and sorghum sourdoughs, respectively. Dextran formation was decreased in favour of oligosaccharide formation when doughs were supplemented with maltose. In conclusion, cold shift fermentation of sourdough with W. cibaria supports high dextran yields or formation of oligosaccharides without excess acidification., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Lactic acid bacteria were isolated from hemp (Cannabis sativa L.) flour, spontaneously fermented dough, and type I sourdough. Isolates were identified and further selected based on pro-technological, nutritional and functional properties. Lactobacillus plantarum/s5, Pediococcus acidilactici/s5, and Leuconostoc mesenteroides/s1 were used as mixed starter to produce hemp sourdough. Significant decreases of the concentration of phytic acid, condensed tannins, and total saponins were observed during fermentation. The in vitro protein digestibility increased up to 90%. Experimental wheat breads were made adding 5% to 15% (w/w) hemp sourdough to the formula, characterized, and compared to baker's yeast wheat bread manufactured without hemp sourdough. The use of hemp sourdough improved the textural features of wheat bread, without adversely affect the sensory profile. Proportionally to the fortification with hemp sourdough, protein digestibility of the breads increased, while the predicted glycemic index significantly decreased (87 vs 100%). This work demonstrated that the fermentation with selected starters improved nutritional functionality of hemp flour, allowing its large-scale use in different food applications, meeting the consumers and producers request for novel fermented baked goods with a well-balanced nutritional profile., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Lactobacilli derive metabolic energy mainly from carbohydrate fermentation. Homofermentative and heterofermentative lactobacilli exhibit characteristic differences in carbohydrate transport and regulation of metabolism, however, enzymes for carbohydrate transport in heterofermentative lactobacilli are poorly characterized. This study aimed to identify carbohydrate active enzymes in the L. reuteri strains LTH2584, LTH5448, TMW1.656, TMW1.112, 100-23, mlc3, and lpuph by phenotypic analysis and comparative genomics. Sourdough and intestinal isolates of L. reuteri displayed no difference in the number and type of carbohydrate-active enzymes encoded in the genome. Predicted sugar transporters encoded by genomes of L. reuteri strains were secondary carriers and most belong to the major facilitator superfamily. The quantification of gene expression during growth in sourdough and in chemically defined media corresponded to the predicted function of the transporters MalT, ScrT and LacS as carriers for maltose, sucrose, and lactose or raffinose, respectively. The genotype for sugar utilization matched the fermentation profile of 39 sugars for L. reuteri strains, and indicated preference for maltose, sucrose, raffinose and (iso)-malto-oligosaccharides, which are available in sourdough and in the upper intestine of rodents. Pentose utilization in L. reuteri species was strain-specific but independent of the origin or phylogenetic position of isolates. Two glycosyl hydrolases, licheninase (EC 3.2.1.73) and endo-1, 4-β-galactosidase (EC 3.2.1.89) were identified based on conserved domains. In conclusion, the study identified the lack of PTS systems, preference for secondary carriers for carbohydrate transport, and absence of carbon catabolite repression as characteristic features of the carbohydrate metabolism in the heterofermentative L. reuteri., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Aiming at meeting the consumers' demand in terms of bio-preservation, the potential of the combination of the lactic acid bacteria fermentation and the addition of hop extract as natural preservative in breadmaking, was exploited. The antifungal properties of a hop (Humulus lupulus) extract were investigated, showing a significant inhibition of the hyphal growth of Aspergillus parasiticus, Penicillium carneum, Penicillium polonicum, Penicillium paneum, Penicillium chermesinum, Aspergillus niger, Penicillium roqueforti. Lactic acid bacteria belonging to species of Enterococcus feacium, Lactobacillus plantarum, Lactobacillus brevis, Lactobacillus helveticus, Lactobacillus curvatus, Pediococcus pentosaceus, and Pediococcus acidilactici were isolated from hop and subjected to selection based on kinetics of growth and acidification. The sourdough (hS) enriched with hop extract (hE), started with three selected strains, had phenols concentration and antioxidant activity higher than those obtained in the same condition but without the hE. Hop-sourdough used in breadmaking delayed the fungal growth (14 days), giving a bread characterized by free aminoacids concentration, antioxidant and phytase activities higher than bread started only with baker's yeast, with or without the addition of hE. Specific volume and cell-total area of the bread containing hE improved, and its sensory profile was characterized by typical sourdough attributes, and a moderate bitter/herbaceous perception.

The microbial ecology of faba bean sourdoughs obtained from an Italian (Ita) and a Finnish (Fi) cultivar, belonging respectively to Vicia faba major and V. faba minor groups, was described by 16S rRNA gene pyrosequencing and culture-dependent analysis. The sourdoughs were propagated with traditional backslopping procedure throughout 14days. Higher microbial diversity was found in the sourdough deriving from V. faba minor (Fi), still containing residual hulls after the milling procedure. After 2days of propagation, the microbial profile of Ita sourdough was characterized by the dominance of the genera Pediococcus, Leuconostoc and Weissella, while the genera Lactococcus, Lactobacillus and Escherichia, as well as Enterobacteriaceae were present in Fi sourdoughs. Yeasts were in very low cell density until the second backslopping and were not anymore found after this time by plate count or pyrosequencing analysis. Among the lactic acid bacteria isolates, Pediococcus pentosaceus, Leuconostoc mesenteroides and Weissella koreensis had the highest frequency of occurrence in both the sourdoughs. Lactobacillus sakei was the only lactobacillus isolated from the first to the last propagation day in Fi sourdough. According to microbiological and acidification properties, the maturity of the sourdoughs was reached after 5days. The presence of hulls and the different microbial composition reflected on biochemical characteristics of Fi sourdoughs, including acidification and phenolic compounds. Moreover, proteolysis in Fi sourdough was more intense compared to Ita. The microbial dynamic of the faba bean sourdoughs showed some differences with the most studied cereal sourdoughs., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Weissella cibaria RBA12 isolated from pummelo from Northeast India produces a dextran composed of 97% α-(1→6) linkages in the main chain and 3% α-(1→3) branched linkages. The in vitro prebiotic activity of dextran-RBA12 was explored. Dextran-RBA12 displayed enhanced growth of probiotic Bifidobacterium and Lactobacillus spp., and controlled growth of non-probiotic enteric bacteria. Dextran-RBA12 showed superior resistance to physiological barriers with a maximum hydrolysis of 0.51%, 0.31% and 0.24% by artificial gastric juice, α-amylase and intestinal fluid, respectively, whereas compared to maximum hydrolysis of 25.23%, 19.13% and 6%, respectively after 5h of incubation shown by commercial prebiotic inulin. The production of dextran from Weissella cibaria RBA12 in sourdough prepared from whole wheat flour, wheat bran and rye bran showed the highest dextran of 3.26±0.12% d.w. in rye bran. The overall study summarized that dextran-RBA12 can be used as a prebiotic and also can be easily produced in sourdough., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Lactic acid bacteria strains, previously isolated from the same matrix, were used to ferment quinoa flour aiming at exploiting the antioxidant potential. As in vitro determined on DPPH and ABTS radicals, the scavenging activity of water/salt-soluble extracts (WSE) from fermented doughs was significantly (P<0.05) higher than that of non-inoculated doughs. The highest inhibition of linoleic acid autoxidation was found for the quinoa dough fermented with Lactobacillus plantarum T0A10. The corresponding WSE was subjected to Reverse Phase Fast Protein Liquid Chromatography, and 32 fractions were collected and subjected to in vitro assays. The most active fraction was resistant to further hydrolysis by digestive enzymes. Five peptides, having sizes from 5 to 9 amino acid residues, were identified by nano-Liquid Chromatography-Electrospray Ionisation-Mass Spectra/Mass Spectra. The sequences shared compositional features which are typical of antioxidant peptides. As shown by determining cell viability and radical scavenging activity (MTT and DCFH-DA assays, respectively), the purified fraction showed antioxidant activity on human keratinocytes NCTC 2544 artificially subjected to oxidative stress. This study demonstrated the capacity of autochthonous lactic acid bacteria to release peptides with antioxidant activity through proteolysis of native quinoa proteins. Fermentation of the quinoa flour with a selected starter might be considered suitable for novel applications as functional food ingredient, dietary supplement or pharmaceutical preparations., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Organic farming is gaining broad recognition as a system that complies well with sustainability, an overarching principle that should drive agriculture now and in the coming year. Different cultivars and products can harbour different abundances of specific bacterial groups, farming system may influence the composition and abundances of microbial communities found on food product. Despite the growing interest towards organic foods, we still have a limited understanding of the diversity of food-associated microbial communities and the factors that influence the composition of these communities. Consumers in developed nations are commonly exposed to differences in farming practices through their choice between organic and conventionally farmed foods. Organic farming practices can differ from conventional farming practices in a variety of ways, including the types of fertilizer and pesticides that are used. This review aiming to gather current knowledge on chemical, technological, toxicological and functional properties and microbiota composition of wheat flours originating from organic and conventional farming systems and how the use of these may affect the sourdough fermentation and breadmaking. Sourdough fermentation is the most natural and best-performing process to ensure optimal sensory and functional characteristics. It fits perfectly into the processing chain that starts with the organic farming, especially for old wheat varieties with weaker technological properties. Recently, organic and sourdough microbiota diversity was investigated and in some case a comparison between organic and conventional microbial ecosystem was also carried out. Opposites evidences arise. Once a higher diversity of lactic acid bacteria species was found in conventional wheat sourdoughs, while when the diversity of Firmicutes was investigated, organic sourdoughs showed the highest complexity. When occurring, the differences between conventional and organic sourdough microbiota and their effects on bread properties are difficult to be identified and categorized due to the extremely large variability in baker's practices. Besides, this review would provide a critical view of this topic in order to avoid the speculation that in this field unavoidably arise., (Copyright © 2016 Elsevier B.V. All rights reserved.)

The market of gluten-free bakery products is considerably growing since better diagnostic methods allow identifying an increasing number of people suffering coeliac disease and other gluten-related disorders such as dermatitis herpetiformis, gluten ataxia, wheat allergy and non-coeliac gluten sensitivity. The only and safe treatment available nowadays for these types of disorders is to follow a strict and permanent lifelong gluten-free diet. Beside the people needing to follow a gluten-free diet for health reasons, a new segment of consumers has arisen who consume gluten-free products as a lifestyle choice. Among the bakery products, bread is a major staple food consumed daily all over the world. The dough and bread quality characteristics (such as gas retaining ability, mixing tolerance, resistance to stretch and extensibility and crumb structure) are mostly attributed to the presence of gluten. Despite the improved quality of gluten-free breads in the last number of years, most products on the market are still described as low quality product. In addition to the low overall quality of gluten-free products, the nutritional value of a large number of them is quite poor. In this context, this review gives an overview on the consumers, which need to follow a gluten-free diet for health reasons. The trends in this gluten-free bakery segment will also be reviewed based on the current analysis of marketing studies. An overview of the major ingredients used in gluten-free bread products will be given. The choice of the ingredients discussed in this paper is based on a comprehensive study of the leading gluten-free breads available on the market, as well as a detailed study of the scientific literature. The impact of the various ingredients on bread-making process and bread quality is also part of this review. Major emphasis will be placed on the application of sourdough as a means to improve gluten-free bread quality., (Copyright © 2016 Elsevier B.V. All rights reserved.)

The drivers for the establishment and composition of the sourdough microbiota, with particular emphasis on lactic acid bacteria, are reviewed and discussed. More than 60 different species of lactobacilli were identified from sourdoughs, showing the main overlapping between sourdough and human intestine ecosystems. The microbial kinetics during sourdough preparation was described by several studies using various methodological approaches, including culture-dependent and -independent (e.g., high throughput sequencing), and metabolite and meta-transcriptome analyses. Although the abundant microbial diversity harbored by flours, a succession of dominating and sub-dominating populations of lactic acid bacteria suddenly occurred during sourdough propagation, leading to the progressive assembly of the bacterial community. The contribution of all the potential sources (house microbiota, flour, types of flours and additional ingredients) for contaminating lactic acid bacteria was compared with the aim to find overlapping or specific routes that affect the sourdough microbiota. Once established and mature, pros and cons regarding the stability of the sourdough lactic acid bacteria biota were also reviewed, showing contradictory results, which were mainly dependent on the species/strains. Probably, the future research efforts should be dedicated to decrease the sources/drivers of noticeable variation rather than to full standardization of the process for sourdough preparation and use., (Copyright © 2016. Published by Elsevier B.V.)

Together with acidifying lactic acid bacteria, yeasts play a key role in the production process of sourdough, where they are either naturally present or added as a starter culture. Worldwide, a diversity of yeast species is encountered, with Saccharomyces cerevisiae, Candida humilis, Kazachstania exigua, Pichia kudriavzevii, Wickerhamomyces anomalus, and Torulaspora delbrueckii among the most common ones. Sourdough-adapted yeasts are able to withstand the stress conditions encountered during their growth, including nutrient starvation as well as the effects of acidic, oxidative, thermal, and osmotic stresses. From a technological point of view, their metabolism primarily contributes to the leavening and flavour of sourdough products. Besides ethanol and carbon dioxide, yeasts can produce metabolites that specifically affect flavour, such as organic acids, diacetyl, higher alcohols from branched-chain amino acids, and esters derived thereof. Additionally, several yeast strains possess functional properties that can potentially lead to nutritional and safety advantages. These properties encompass the production of vitamins, an improvement of the bioavailability of phenolic compounds, the dephosphorylation of phytic acid, the presence of probiotic potential, and the inhibition of fungi and their mycotoxin production. Strains of diverse species are new candidate functional starter cultures, offering opportunities beyond the conventional use of baker's yeast., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Three obligately heterofermentative lactic acid bacteria (LAB) strains (Lactobacillus sanfranciscensis PON100336, Leuconostoc citreum PON10079 and Weissella cibaria PON10030) were used in this study as a multi-species starter culture for sourdough production. The starter inoculum was prepared and propagated in sterile semolina extract (SSE) broth. Acidification kinetics, microbiological counts detected on specific media for sourdough LAB, polymorphic profile comparison and species-specific PCRs evidenced a stability of the liquid inoculum over time determining its suitability for direct addition to semolina. In order to validate this innovative method for the production of durum wheat (Triticum durum Desf) sourdoughs, 15 semolinas (from ten old and five modern genotypes cultivated in Sicily, southern Italy) were used to prepare the SSEs and to produce sourdoughs and finally breads. Chemical and microbiological analyses of the sourdoughs and the evaluation of the quality parameters (weight loss, height, crumb and crust colour, image analysis and volatile organic compound generation) of the resulting breads indicated that the direct addition of the liquid inocula propagated in SSE is a valuable method to stabilise the production of sourdoughs. The differences registered during the technological characterisation of the breads were underlined by the sensory tests and the multivariate analysis and are mainly imputable to the type of semolina., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Exopolysaccharides from lactic acid bacteria may improve texture and shelf life of bread. The effect of exopolysaccharides on bread quality, however, depends on properties of the EPS and the EPS producing strain. This study investigated structure-function relationships of EPS in baking application. The dextransucrase DsrM and the reuteransucrase GtfA were cloned from Weissella cibaria 10M and Lactobacillus reuteri TMW1.656, respectively, and heterologously expressed in Escherichia coli. Site-directed mutagenesis of GtfA was generates reuterans with different glycosidic bonds. NMR spectrum indicated reuteranPI, reuteranNS and reuteranPINS produced by GtfA-V1024P:V1027I, GtfA-S1135N:A1137S and GtfA-V1024P:V1027I:S1135N:A1137S, respectively, had a higher proportion of α-(1→4) linkages when compared to reuteran. ReuteranNS has the lowest molecular weight as measured by asymmetric flow-field-flow fractionation. The reuteransucrase negative mutant L. reuteri TMW1.656ΔgtfA was generated as EPS-negative derivative of L. reuteri TMW1.656. Cell counts, pH, and organic acid levels of sourdough fermented with L. reuteri TMW1.656 and TMW1.656ΔgtfA were comparable. Reuteran produced by L. reuteri TMW1.656 during growth in sourdough and reuteran produced ex situ by GtfA-ΔN had comparable effects on bread volume and crumb hardness. Enzymatically produced dextran improved volume and texture of wheat bread, and of bread containing 20% rye flour. ReuteranNS but not reuteranPI or reuteran was as efficient as dextran in enhancing wheat bread volume and texture. Overall, reuteran linkage type and molecular weight are determinants of EPS effects on bread quality. This study established a valuable method to elucidate structure-function relationships of glucans in baking applications., (Copyright © 2016 Elsevier B.V. All rights reserved.)

A simple and specific method for the rapid detection and identification of Lactobacillus brevis was developed. A fAFLP (Fluorescent Amplified Fragment Length Polymorphisms) marker for L. brevis was used to design oligonucleotide primers for a species-specific PCR assay, targeting a 125bp fragment of the gene encoding the aldo/keto reductase of the diketogulonate-reductase family of L. brevis. This assay resulted in 100% inclusivity and exclusivity of assignment of strains to the species L. brevis. The analytical specificity of this assay was successfully tested to identify L. brevis isolates from sourdoughs., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Barley flour is not commonly used for baking because of its negative effects on bread dough rheology and loaf volume. However, barley sourdoughs are promising ingredients to produce improved barley-based breads. Spontaneous barley sourdough fermentations were performed through backslopping (every 24h, 10days) under laboratory (fermentors, controlled temperature of 30°C, high dough yield of 400) and bakery conditions (open vessels, ambient temperature of 17-22°C, low dough yield of 200), making use of the same batch of flour. They differed in pH evolution, microbial community dynamics, and lactic acid bacteria (LAB) species composition. After ten backsloppings, the barley sourdoughs were characterized by the presence of the LAB species Lactobacillus fermentum, Lactobacillus plantarum, and Lactobacillus brevis in the case of the laboratory productions (fast pH decrease, pH<4.0 after two backslopping steps), and of Leuconostoc citreum, Leuconostoc mesenteroides, Weissella confusa and Weissella cibaria in the case of the bakery productions (slow pH decrease, pH4.0 after eight backslopping steps). In both sourdough productions, Saccharomyces cerevisiae was the sole yeast species. Breads made with wheat flour supplemented with 20% (on flour basis) barley sourdough displayed a firmer texture, a smaller volume, and an acceptable flavour compared with all wheat-based reference breads. Hence, representative strains of the LAB species mentioned above, adapted to the environmental conditions they will be confronted with, may be selected as starter cultures for the production of stable barley sourdoughs and flavourful breads., (Copyright © 2016. Published by Elsevier B.V.)

Lactobacillus sanfranciscensis is the predominant key lactic acid bacterium in traditionally fermented sourdoughs. Despite its prevalence, sourdough and their related breads could be different regarding their physicochemical and sensorial characteristics. The intraspecific diversity of L. sanfranciscensis might explain these observations. Fifty-nine strains isolated from French sourdoughs were typed by a polyphasic approach including Multilocus Sequence Typing (MLST) and Pulsed-field Gel Electrophoresis (PFGE), in order to study their genotypic diversity. MLST scheme can be reduced from six to four gene fragments (gdh, gyrA, nox and pta) without a major loss of discrimination between strains. The genes mapA and pgmA are not good candidates for inclusion in an MLST scheme to type L. sanfranciscensis strains, as they could not be amplified for a set of 18 strains among the 59 studied. This method revealed 20 sequence types (STs). Of these, 19 STs were grouped in one clonal complex, showing a strong relatedness between these strains. PFGE using SmaI discriminated 41 pulsotypes and so distinguished isolates better than the MLST scheme. Both genotypic methods indicate a low diversity between strains isolated from the same sourdough and a higher diversity between strains isolated from different sourdoughs, suggesting an influence of baker practices and/or environmental conditions on the selection of strains. The use of these two methods targeting genetic variations gives an optimal genotypic characterization of L.sanfranciscensis strains., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Wheat bread is the most popular staple food consumed in Tunisia and, despite the niche production of some typical breads (e.g. Tabouna, Mlawi, Mtabga), the major part is currently produced with baker's yeast at industrial or, mainly, at artisanal level, while the use of sourdough fermentation is rarely reported. Considering the growing national demand for cereal baked goods, it can be hypothesized that sourdough fermentation through the use of selected lactic acid bacteria as starters could improve the overall quality and the diversification of local products. Different cereal grains were collected from the regions of Ariana, Bizerta, Beja Nabeul, and Seliana, and the autochthonous lactic acid bacteria were isolated, identified, characterized and selected on the basis of the kinetics of acidification, the proteolytic activity, and the quotient of fermentation. Lactobacillus curvatus MA2, Pediococcus pentosaceus OA2, and Pediococcus acidilactici O1A1 were used together as mixed starter to obtain a selected sourdough. According to the backslopping procedure, a type I sourdough was made from a Tunisian flour (spontaneous sourdough). Compared to the use of the spontaneous sourdough, the one obtained with selected and mixed starters by a unique fermentation step, favored the increase of the concentrations of organic acids, phenols, and total free amino acids, the most suitable quotient of fermentation, and the most intense phytase and antioxidant activities, that increased ca. 20% compared to the control. Moreover, the selected starters improved the in vitro protein digestibility (ca. 82% when selected sourdough was used), textural and sensory features of the breads, as determined by textural profile analysis and panel test, respectively. This study aimed at exploiting the potential of selected autochthonous lactic acid bacteria and extending the use of a sourdough (type II), thanks to the set-up of a two-step fermentation protocol designed for application at the industrial level, and the confirmed nutritional, textural, and sensory advantages of the final product., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Phytases are enzymes capable of sequentially dephosphorylating phytic acid to products of lower chelating capacity and higher solubility, abolishing its inhibitory effect on intestinal mineral absorption. Genetic constructions were made for expressing two phytases from bifidobacteria in Lactobacillus casei under the control of a nisin-inducible promoter. L. casei was able of producing, exporting and anchoring to the cell wall the phytase of Bifidobacterium pseudocatenulatum. The phytase from Bifidobacterium longum spp. infantis was also produced, although at low levels. L. casei expressing any of these phytases completely degraded phytic acid (2mM) to lower myo-inositol phosphates when grown in MRS medium. Owing to the general absence of phytase activity in lactobacilli and to the high phytate content of whole grains, the constructed L. casei strains were applied as starter in a bread making process using whole-grain flour. L. casei developed in sourdoughs by fermenting the existing carbohydrates giving place to an acidification. In this food model system the contribution of L. casei strains expressing phytases to phytate hydrolysis was low, and the phytate degradation was mainly produced by activation of the cereal endogenous phytase as a consequence of the drop in pH. This work shows the capacity of lactobacilli to be modified in order to produce enzymes with relevance in food technology processes. The ability of these strains in reducing the phytate content in fermented food products must be evaluated in further models., (Copyright © 2015 Elsevier B.V. All rights reserved.)

Sixteen sourdoughs (FS1-FS16) used for the manufacture of traditional French breads were characterized by strongly acid conditions (median value of pH 3.5). The concentration of free amino acids (FAA) was highly variable, due to different proteolytic activity of flour used for back slopping and of dominant microorganisms. Median value of cell density of lactic acid bacteria (LAB) was 9.2 log CFU/g. The ratio between LAB and yeasts ranged from 10,000:1 to 10:1. According to the culture-dependent method and 16S metagenetics, Lactobacillus sanfranciscensis was the dominant species in French sourdoughs. FS5 and FS15, propagated according to protocols including one back slopping step at 14 °C, were the only exceptions. High positive correlations were found between L. sanfranciscensis, temperature of back slopping and FAA. The results of this study highlighted the broad adaptability of L. sanfranciscensis to very acid sourdough. Besides species frequently encountered (e.g., Lactobacillus parabrevis/Lactobacillus hammesii, Lactobacillus plantarum and Leuconostoc mesenteroides), first Lactobacillus xiangfangensis (FS5) and Lactobacillus diolivorans (FS15) were found in sourdough. As determined by RAPD-PCR analyses, the sourdough samples showed a different number of strains, ranging from 5 (FS9, FS11 and FS15) to 12 (FS1 and FS13), meaning a highly variable bacterial diversity. Cluster analysis showed that different sourdoughs, especially when propagated in the same bakery, may harbor similar strains. Except for L. plantarum (FS5) and Ln. mesenteroides (FS3), all the dominant species were detected by both 16S metagenetics and culture-dependent method. Yeast diversity was lower than LAB. Except for FS4 (solely dominated by Kazachstania servazzii), yeast microbiota of French sourdoughs was dominated by Saccharomyces cerevisiae. Strains isolated in this study could be a useful base for developing new basic researches on physiology, metabolism, and intraspecific diversity of L. sanfranciscensis, as well as for standardizing the quality of traditional French breads., (Copyright © 2015 Elsevier B.V. All rights reserved.)

Fourteen bakeries located in different regions of France were selected. These bakers use natural sourdough and organic ingredients. Consequently, different organic sourdoughs used for the manufacture of French bread were studied by the enumeration of lactic acid bacteria (LAB) and 16S rRNA sequencing of the isolates. In addition, after DNA extraction the bacterial diversity was assessed by pyrosequencing of the 16S rDNA V1-V3 region. Although LAB counts showed significant variations (7.6-9.5log10CFU/g) depending on the sourdough studied, their identification through a polyphasic approach revealed a large predominance of Lactobacillus sanfranciscensis in all samples. In ten sourdoughs, both culture and independent methods identified L. sanfranciscensis as the dominant LAB species identified. In the remaining sourdoughs, culture methods identified 30-80% of the LAB as L. sanfranciscensis whereas more than 95% of the reads obtained by pyrosequencing belonged to L. sanfranciscensis. Other sub-dominant species, such as Lactobacillus curvatus, Lactobacillus hammesii, Lactobacillus paralimentarius, Lactobacillus plantarum, Lactobacillus pentosus, and Lactobacillus sakei, were also identified. Quantification of L. sanfranciscensis by real-time PCR confirmed the predominance of this species ranging from 8.24 to 10.38log10CFU/g. Regarding the acidification characteristics, sourdough and related bread physico-chemical characteristics varied, questioning the involvement of sub-dominant species or L. sanfranciscensis intra-species diversity and/or the role of the baker's practices., (Copyright © 2015 Elsevier B.V. All rights reserved.)

While phages of lactobacilli are extensively studied with respect to their structure and role in the dairy environment, knowledge about phages in bacteria residing in sourdough fermentation is limited. Based on the previous finding that the Lactobacillus sanfranciscensis phage EV3 carries a putative dextranase gene (dex), we have investigated the distribution of similar dex(+) phages in L. sanfranciscensis, the chance of gene transfer and the properties of the dextranase encoded by phage EV3. L. sanfranciscensis H2A (dex(-)), originally isolated from a wheat sourdough, expressed a Dex(+) phenotype upon infection with EV3. The dextranase gene was isolated from the transductant and heterologously expressed in Escherichia coli. The gene encoded a protein of 801 amino acids with a calculated molecular weight (Mw) of 89.09 kDa and a calculated pI of 5.62. Upon purification aided by a 6-His tag, enzyme kinetic parameters were determined. The Km value was 370 mM, and the Vmax was calculated in about 16 μmol of glucose released from dextran by 1 mg of enzyme in 1 min in a buffer solution at pH 5.0. The optimum conditions were 60 °C and pH 4.5. The enzyme retained its activity for >3h at 60 °C and exhibited only 40% activity at 30 °C; the highest homology of 72% was found to a dextranase gene from Lactobacillus fermentum phage φPYB5. Within 25 L. sanfransiscensis isolates tested, the strain 4B5 carried a similar prophage encoding a dextranase gene. Our data suggest a phage-mediated transfer of dextranase genes in the sourdough environment resulting in superinfection-resistant L. sanfranciscensis Dex(+) strains with a possible ecological advantage in dextran-containing sourdoughs., (Copyright © 2015 Elsevier B.V. All rights reserved.)

Lactobacillus sanfranciscensis is a bacterium used in sourdough that provides desirable properties such as better flavor and texture to the sourdough bread. Here, the intra-species diversity of L. sanfranciscensis strains isolated from Korean sourdough was studied using genotypic (multiplex-RAPD-PCR: multiplex-Randomly Amplified Polymorphic DNA-polymerase chain reaction) and phenotypic (VITEK2 Compact system) analyses. For this, a novel species-specific set of PCR primers was developed to identify L. sanfranciscensis using the recently published genome database. The primers were able to detect L. sanfranciscensis isolated from Korean sourdough with 100% accuracy. Genotyping and phenotyping analyses at the strain level demonstrated that Korean sourdough possesses various biotypes of L. sanfranciscensis strains. These strains were clustered into 5 subtypes (genotyping) or 7 subtypes (phenotyping). In summary, this strategy to construct novel primers reduced the chance of cross amplification and was able to identify the desired strain. The various strains isolated in this study can be used to develop a sourdough starter after the analysis of their fermentation characteristics., (Copyright © 2015 Elsevier B.V. All rights reserved.)

This study aimed at evaluating the composition of nineteen traditional Italian legumes and at investigating the potential of the sourdough fermentation with selected lactic acid bacteria to improve the nutritional and functional features. Traditional Italian legumes, all with product certifications and belonging to Phaseolus vulgaris, Cicer arietinum, Lathyrus sativus, Lens culinaris and Pisum sativum species, were used in this study. Seeds were milled, and flours were analyzed for proximate composition and subjected to sourdough fermentation at 30°C for 24h. Lactobacillus plantarum C48 and Lactobacillus brevis AM7 were used as selected starters. Compared to control doughs, without bacterial inoculum, the concentrations of free amino acids (FAA), soluble fibres, and total phenols increased for all legume sourdoughs. Raffinose decreased of up to ca. 64%. During sourdough fermentation, the level of GABA markedly increased and reached values up to 624mg/kg. Condensed tannins decreased. At the same time, almost all legume sourdoughs showed increases of the antioxidant and phytase activities. As shown by PCA analysis based on data of total FAA, GABA, raffinose, soluble/insoluble dietary fibre, condensed tannins and antioxidant and phytase activities, all legume sourdoughs were clearly differentiated from control doughs. The traditional Italian legumes are bio-diverse, and all showed high levels of nutritional elements and suitability for optimal sourdough fermentation. Legume flours subjected to sourdough fermentation would be suitable to be used alone or better in mixture with cereals, and as gluten-free ingredients for making novel and healthy foods., (Copyright © 2014 Elsevier B.V. All rights reserved.)

This study aimed at investigating the addition of legume (chickpea, lentil and bean) flours to wheat flour bread. Type I sourdough containing legumes or wheat-legume flours were prepared and propagated (back slopped) in laboratory, according to traditional protocols that are routinely used for making typical Italian breads. Based on kinetic of acidification and culture-dependent data, the wheat-legume sourdough was further characterized and selected for bread making. As determined by RAPD-PCR and partial sequencing of 16S rDNA gene analyses, lactic acid bacteria in wheat-legume sourdough included Lactobacillus plantarum, Lactobacillus sanfranciscensis, Leuconostoc mesenteroides, Lactobacillus fermentum, Weissella cibaria, Lactobacillus pentosus, Lactobacillus coryneformis, Lactobacillus rossiae, Lactobacillus brevis, Lactobacillus parabuchneri and Lactobacillus paraplantarum. Two breads containing 15% (w/w) of legume (chickpea, lentil and bean) flours were produced using selected wheat-legume sourdough (WLSB) and traditional wheat sourdough (WSB). Compared to wheat yeasted bread (WYB), the level of total free amino acids (FAA) was higher in WSB and WLSB. Phytase and antioxidant activities were the highest in WLSB. Compared to bread WYB, the addition of legume flours decreased the in vitro protein digestibility (IVPD) (WYB versus WSB). However, the dough fermentation with WSLB favored an increase of IVPD. According to the levels of carbohydrates, dietary fibers and resistant starch, WSB and WLSB showed lower values of hydrolysis index (HI) compared to WYB. As showed by texture and image analyses and sensory evaluation of breads, a good acceptability was found for WSB and, especially, WLSB breads., (Copyright © 2014 Elsevier B.V. All rights reserved.)