New Insights into the Oenological Significance of Candida zemplinina: Impact of Selected Autochthonous Strains on the Volatile Profile of Apulian Wines.

In this investigation, we explored the oenological significance of Candida zemplinina (syn. Starmerella bacillaris) isolates from Apulian grape musts. Moreover, we provide the first evidence of the impact of different C. zemplinina strains on the wine aromatic properties tested as monocultures. We described the diversity of C. zemplinina strains isolated from grapes and the variability of 'volatile' phenotypes associated with this intraspecific variability. Thirty-three isolates were characterized at strain level by PCR-based approach and, among these, 16 strains were identified and then tested by microfermentation tests carried out in grape must. Analyzed strains were low producers of acetic acid and hydrogen sulphide, not able to decarboxylate a panel of representative amino acids, whereas they showed fructophilic character and significant glycerol production. Volatile profiles of produced wines were investigated by gas chromatography-mass spectrometry. The Odor Activity Values of all molecules were calculated and 12 compounds showed values above their odor thresholds. Two selected strains (35NC1 and 15PR1) could be considered as possible starter cultures since they were able to positively affect the sensory properties of obtained wine. This report firstly supplies evidence on the strain-specific impact of different C. zemplinina strains on the final aroma of produced wines.

Selection of non-Lactobacillus strains to be used as starters for sourdough fermentation.

The suitability of forty-one non-Lactobacillus strains to be used as selected starters for sourdough fermentation was evaluated. According to the data collected, Pediococcus pentosaceus OA1 and S3N3 and Leuconostoc citreum PRO17 were selected based on the optimal acidification and growth performances and the intense proteolytic activity (increase of TFFA up to 80%) on whole wheat flour doughs. A relevant degradation of phytic acid (up to 58%) and the increase of phenols content and scavenging activity (4- and 2-folds, respectively) were also observed. The technological performances were compared to two representative Lactobacillus strains (Lactobacillus plantarum and Lactobacillus sanfranciscensis). The investigation of the robustness of the selected strains during the propagation (back-slopping procedure) showed their long-term dominance only when singly-inoculated; while Leuc. citreum PRO17 dominated the fermentation when the strains were co-inoculated. The sourdoughs obtained by the non-Lactobacillus selected strains (singly or pooled) were used for breadmaking. Selected sourdoughs allowed the production of breads characterized by in-vitro protein digestibility (IVPD) higher than that of breads obtained with Lactobacillus strains or baker's yeast. The aroma profile, estimated by GC/MS, was complex and characterized by high concentration of the typical compounds (hexanol, 3-methylbutanol and 2-pentylfuran) of sourdough bread.

Wasted Bread as Substrate for the Cultivation of Starters for the Food Industry.

The amount of bread wasted daily worldwide, throughout its entire lifecycle, from production to distribution, is estimated to be hundreds of tons, therefore representing both economic and environmental issues. This work aimed at the valorization of wasted bread, setting-up a protocol for obtaining a growth medium to be used for the cultivation of food industry microbial starters. The optimization of the protocol included the set-up of parameters for the hydrolysis of the bread nutrient compounds with proteolytic and amylolytic enzymes and the supplementation with nitrogen and/or carbon sources. The suitability of the optimized medium for the growth of lactic acid bacteria, yeasts and fungi from dairy, bakery, and wine industries was assessed. Lactic acid bacteria growth was strongly affected by the quantity and quality of nitrogen sources employed, while yeasts and fungi growth exceeded that obtained with the reference media commonly employed for their cultivation. Wasted bread medium (WBM) represents a realistic option for the valorization and re-use of bread waste, responding to the modern vision of circular economy.

Lactic Acid Fermentation as a Pre-Treatment Process for Faba Bean Flour and Its Effect on Textural, Structural and Nutritional Properties of Protein-Enriched Gluten-Free Faba Bean Breads.

Lactic acid fermentation could be used as a potential modification tool for faba bean flour to enable its incorporation in boosting the nutritional profile of gluten-free breads. Gluten-free breads made with fermented or unfermented faba bean flours were compared with commercial soy flour. The amounts of faba- and soy-bean flours were adjusted to obtain the same protein content in bread (16%). Both fermented and unfermented faba bean flour resulted in larger bread volume (2.1 mL/g and 2.4 mL/g, respectively) compared to bread made with soybean flour (1.5 mL/g). Breads made with unfermented and fermented faba flour had higher porosity (82% and 72%, respectively) than bread with soy flour (61%). The faba breads also were softer than the soy bread. Fermentation of faba flour prior to bread making significantly increased crumb hardness (584 vs. 817 g). Fermentation increased in vitro protein digestibility (72.3% vs. 64.8%). Essential Amino Acid and Biological Value indexes were significantly higher for breads containing fermented faba flour compared to breads made with unfermented faba and soy flour. The Protein Efficiency Ratio and Nutritional Index increased by fermentation from 33 to 36 and 1.6 to 2.7, respectively. Pre-fermentation of faba bean flour improved the nutritional properties of high-protein, gluten-free faba bread. A sensory panel indicated that fermentation did not affect the crumbliness, evenness of pore size and springiness of breadcrumb.

Characterization of indigenous Pediococcus pentosaceus, Leuconostoc kimchii, Weissella cibaria and Weissella confusa for faba bean bioprocessing.

The interest towards legumes in food applications has risen over the past decades. However, the presence of antinutritional factors (ANF) and the poor technological performances still restricts their application in food fortification. In this study, four lactic acid bacteria (LAB) isolated from faba bean were applied as starter cultures for faba bean bioprocessing. None of the strains employed produced exopolysaccharides from raffinose, on the contrary, they did with sucrose as substrate. The fermented doughs were characterized and the strains were compared for their adaptation capacity and metabolic performance including the formation of dextrans, the degradation of ANF and the ability to improve antioxidant activity and in vitro protein digestibility (IVPD). A contribution to the proteolysis was given by the presence of endogenous enzymes, responsible for the increase of peptides and amino acids in dough from irradiated flour. However, the LAB strains further enhanced proteolysis. Weissella cibaria VTT E-153485 led to the highest peptide release and consequentially to the highest IVPD. In doughs fermented with Pediococcus pentosaceus VTT E-153483 and Leuconostoc kimchi VTT E-153484, phytic acid was reduced to more than half the initial concentration. Inoculated doughs had significantly lower content of oligosaccharides after 24 h of incubation compared to the controls. The most efficient raffinose consumption was found for Leuc. kimchi and W. cibaria. Doughs inoculated with weissellas contained >1% of dextrans. Weissella confusa VTT E-143403 induced a significant increment in viscosity (ca. 7 times higher than the controls). This study revealed that well-characterized, indigenous LAB provided beneficial biotechnological features in faba bean dough processing and contributed to its implementation in the food production.

Performance of Leuconostoc citreum FDR241 during wheat flour sourdough type I propagation and transcriptional analysis of exopolysaccharides biosynthesis genes.

This study focused on the performance of the dextran producer Leuconostoc citreum as starter culture during 30 days of wheat flour type I sourdough propagation (back-slopping). As confirmed by RAPD-PCR analysis, the strain dominated throughout the propagation procedure, consisting of daily fermentations at 20 °C. The sourdoughs were characterized by consistent lactic acid bacteria cell density and acidification parameters, reaching pH values of 4.0 and mild titratable acidity. Carbohydrates consumption remained consistent during the propagation procedure, leading to formation of mannitol and almost equimolar amount of lactic and acetic acid. The addition of sucrose enabled the formation of dextran, inducing an increase in viscosity of the sourdough of 2-2.6 fold, as well as oligosaccharides. The transcriptional analysis based on glucosyltransferases genes (GH70) showed the existence in L. citreum FDR241 of at least five different dextransucrases. Among these, only one gene, previously identified as forming only α-(1-6) glycosidic bonds, was significantly upregulated in sourdough fermentation conditions, and the main responsible of dextran formation. A successful application of a starter culture during long sourdough back-slopping procedure will depend on the strain robustness and fermentation conditions. Transcriptional regulation of EPS-synthetizing genes might contribute to increase the efficiency of industrial processes.

Influence of fermented faba bean flour on the nutritional, technological and sensory quality of fortified pasta.

Faba bean has gained increasing attention from the food industry and the consumers mainly due to the quality of its protein fraction. Fermentation has been recently recognized as the most efficient tool for improving its nutritional and organoleptic properties. In this study, faba bean flour fermented with Lactobacillus plantarum DPPMAB24W was used to fortify semolina pasta. Pasta samples including different percentages of fermented faba bean flour were produced at the pilot-plant level and characterized using an integrated approach for chemical, nutritional, technological, and sensory features. At a substitution level of 30%, pasta had a more homogeneous texture and lower cooking loss compared to 50% addition. The impact of faba bean flour addition on pasta technological functionality, particularly of the protein fraction, was also assessed by scanning electron microscopy and textural profile analysis. Compared to traditional (semolina) pasta and pasta containing unfermented faba bean flour, the nutritional profile (in vitro protein digestibility and nutritional indexes - chemical score (CS), sequence of limiting essential amino acids, Essential Amino Acid Index (EAAI), Biological Value (BV), Protein Efficiency Ratio (PER), and Nutritional Index (NI)) and the resistant starch content of pasta containing 30% fermented faba bean flour markedly improved, while the starch hydrolysis rate decreased, without negatively affecting technological and sensory features. The use of fermentation technology appears to be a promising tool to enhance the quality of pasta and to promote the use of faba bean flour.

Exploring the Microbiota of Faba Bean: Functional Characterization of Lactic Acid Bacteria.

This study investigated the metabolic traits of 27 lactic acid bacteria (LAB) strains belonging to different species, previously isolated from faba bean. The activities assayed, related to technological and nutritional improvement of fermented faba bean, included peptidases, ß-glucosidase, phytase, as well as exopolysaccharides synthesis and antimicrobial properties. In addition, the bacteria performance as starter cultures during faba bean fermentation on proteolysis, antioxidant potential, and degradation of condensed tannins were assessed. Fermentative profiling showed that only 7 out of 27 strains were able to metabolize D-raffinose, particularly Leuc. mesenteroides I01 and I57. All strains of Pediococcus pentosaceus exerted high PepN activity and exhibited ß-glucosidase activity higher than the median value of 0.015 U, while phytase activity was largely distributed among the different strains. All the weissellas, and in lower amount leuconostocs, showed ability to produce EPS from sucrose. None of the strains did not survive the simulated gastrointestinal tract with the exception of P. pentosaceus I56, I76, 147, I214, having a viability of 8-9 log CFU/ml at the end of the treatment. None of the strains showed antimicrobial activity toward Staphylococcus aureus, while eight strains of P. pentosaceus exhibited a strong inhibitory activity toward Escherichia coli and Listeria monocytogenes. Generally, the doughs fermented with pediococci exhibited high amount of total free amino acids, antioxidant activity, and condensed tannins degradation. These results allowed the identification of LAB biotypes as potential starter cultures for faba bean bioprocessing, aiming at the enhancement of faba bean use in novel food applications.

How organic farming of wheat may affect the sourdough and the nutritional and technological features of leavened baked goods.

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.

Impact of Enzymatic and Microbial Bioprocessing on Protein Modification and Nutritional Properties of Wheat Bran.

Besides providing dietary fiber, wheat bran is a recognized source of protein and is considered a very valuable substitute for other protein-rich sources in the food and feed industry. Nonetheless, several factors affect protein bioavailability, including bran's layered structure. This study showed the influence on the release and protein modification of wheat bran of different bioprocessing methods involving the activation of endogenous enzymes of bran, the addition of an enzyme mixture having carbohydrase activity, and microbial fermentation. Bioprocessing in acidic conditions significantly enhanced the solubilization of protein from wheat bran, reaching the highest value in the treatment where the sole endogenous protease activity was activated. Bioprocessing through controlled fermentation allowed a more intense proteolysis and strongly impacted the in vitro digestibility of proteins. The combined use of starter cultures and cell-wall-degrading enzymes was characterized by the highest increase of phytase activity and total phenols.

Quorum-sensing regulation of constitutive plantaricin by Lactobacillus plantarum strains under a model system for vegetables and fruits.

This study aimed at investigating the regulatory system of bacteriocin synthesis by Lactobacillus plantarum strains in vegetables and fruits in a model system. Sterile and neutralized cell-free supernatant (CFS) from L. plantarum strains grown in MRS broth showed in vitro antimicrobial activities toward various indicator strains. The highest activity was that of L. plantarum C2. The antimicrobial activity was further assayed on vegetable and fruit agar plates (solid conditions) and in juices (liquid conditions). A regulatory mechanism of bacteriocin synthesis via quorum sensing was hypothesized. The synthesis of antimicrobial compounds seemed to be constitutive under solid conditions of growth on vegetable and fruit agar plates. In contrast, it depended on the size of the inoculum when L. plantarum C2 was grown in carrot juice. Only the inoculum of ca. 9.0 log CFU ml(-1) produced detectable activity. The genes plnA, plnEF, plnG, and plnH were found in all L. plantarum strains. The genes plnJK and plnN were detected in only three or four strains. Reverse-phase high-performance liquid chromatography purification and mass spectrometry analysis revealed the presence of a mixture of eight peptides in the most active fraction of the CFS from L. plantarum C2. Active peptides were encrypted into bacteriocin precursors, such as plantaricins PlnJ/K and PlnH and PlnG, which are involved in the ABC transport system. A real-time PCR assay showed an increase in the expression of plnJK and plnG during growth of L. plantarum C2 in carrot juice.

How the sourdough may affect the functional features of leavened baked goods.

Sourdough fermentation is one of the oldest food biotechnologies, which has been studied and recently rediscovered for its effect on the sensory, structural, nutritional and shelf life properties of leavened baked goods. Acidification, proteolysis and activation of a number of enzymes as well as the synthesis of microbial metabolites cause several changes during sourdough fermentation, which affect the dough and baked good matrix, and influence the nutritional/functional quality. Currently, the literature is particularly rich of results, which show how the sourdough fermentation may affect the functional features of leavened baked goods. In the form of pre-treating raw materials, fermentation through sourdough may stabilize or to increase the functional value of bran fractions and wheat germ. Sourdough fermentation may decrease the glycaemic response of baked goods, improve the properties and bioavailability of dietary fibre complex and phytochemicals, and may increase the uptake of minerals. Microbial metabolism during sourdough fermentation may also produce new nutritionally active compounds, such as peptides and amino acid derivatives (e.g., γ-amino butyric acid) with various functionalities, and potentially prebiotic exo-polysaccharides. The wheat flour digested via fungal proteases and selected sourdough lactobacilli has been demonstrated to be probably safe for celiac patients.

Exploitation of the health-promoting and sensory properties of organic pomegranate (Punica granatum L.) juice through lactic acid fermentation.

Two strains (POM1 and C2) or LP09 of Lactobacillus plantarum, which were previously isolated from tomatoes and carrots, and another commercial strain of L. plantarum (LP09), were selected to singly ferment (30 °C for 120 h) pomegranate juice (PJ) under standardized protocol. PJs were further stored at 4 °C for 30 days. Filtered PJ, not added of starters (unstarted PJ), was used as the control. After fermentation, all starters grew to ca. 9.0 Log CFU/mL. Viable cells of strain LP09 sharply decreased during storage. The other two strains survived to ca. 7.0 and 8.0 Log CFU/mL. Lactic acid bacteria consumed glucose, fructose, malic acid, and branched chain and aromatic amino acids. The concentration of free fatty acids increased for all started PJs. Compared to unstarted PJ, color and browning indexes of fermented PJs were preferable. The concentration of total polyphenolic compounds and antioxidant activity were the highest for started PJs, with some differences that depended on the starter used. Fermentation increased the concentration of ellagic acid, and enhanced the antimicrobial activity. Fermented PJs scavenged the reactive oxygen species generated by H2O2 and modulated the synthesis of immune-mediators from peripheral blood mononuclear cells (PBMC). Unstarted and fermented PJs inhibited the growth of K562 tumor cells. The sensory attributes of fermented PJs were preferred. The fermentation of pomegranate juice would represent a novel technology option, which joins health-promoting, sensory and preservative features to exploit the potential of pomegranate fruits.

Antifungal activity of Meyerozyma guilliermondii: identification of active compounds synthesized during dough fermentation and their effect on long-term storage of wheat bread.

Preliminarily, 146 strains of yeasts were screened for the antifungal activity toward the indicator Penicillium roqueforti DPPMAF1. The strain Meyerozyma guilliermondii LCF1353 was selected and used for dough fermentation. The water/salt soluble extract of the dough was analyzed by HPLC and GC/MS-SPME. The synthesis of the extracellular cell wall-degrading enzyme ß-1,3-glucanase and ethyl-acetate was shown. The effect on conidia germination mainly suggested a fungistatic activity. M. guilliermondii LCF1353 was used as starter for dough fermentation in combination with Wickeramomyces anomalus 1695 and Lactobacillus plantarum 1A7, which were previously selected for antifungal activity. The growth of the strains was monitored by plate count and molecular techniques, and competitive or antagonistic interactions among them were excluded. Bread started with the combination of M. guilliermondii LCF1353, W. anomalus LCF1695 and L. plantarum 1A7 showed a more prolonged shelf life compared to the other breads. Fungal growth was delayed at least until 14 days of storage, under conditions of high artificial inoculum. The bread manufactured with the above combination showed good chemical and textural characteristics and, as shown by sensory analysis, it was appreciated for elasticity, color and taste.

Synthesis of the cancer preventive peptide lunasin by lactic acid bacteria during sourdough fermentation.

This study aimed to exploit the potential of sourdough lactic acid bacteria to release lunasin during fermentation of cereal and nonconventional flours. The peptidase activities of a large number of sourdough lactic acid bacteria were screened using synthetic substrates. Selected lactic acid bacteria were used as sourdough starters to ferment wholemeal wheat, soybean, barley, amaranth, and rye flours. Proteinase activity during fermentation was characterized by SDS-PAGE analysis of the water-soluble extracts. Albumins having molecular masses of 18 to 22 kDa, which included the size of lunasin precursors, were markedly affected by proteolysis of lactic acid bacteria. After fermentation, lunasin from the water-soluble extracts was quantified, purified, and identified through RP-HPLC and nano-LC-ESI-MS analyses. Compared to control doughs, the concentration of lunasin increased up to 2-4 times during fermentation. Lactobacillus curvatus SAL33 and Lactobacillus brevis AM7 synthesized the highest concentrations of lunasin in all the flours. Besides the presence of the entire lunasin sequence, fragments containing the immunoreactive epitope RGDDDDDDDDD were also found. This study shows that fermentation by lactic acid bacteria increased the concentration of lunasin to levels that would suggest new possibilities for the biological synthesis and for the formulation of functional foods.

Effect of lactic acid fermentation on antioxidant, texture, color and sensory properties of red and green smoothies.

Weissella cibaria, Lactobacillus plantarum, Lactobacillus sp. and Lactobacillus pentosus were variously identified from blackberries, prunes, kiwifruits, papaya and fennels by partial 16S rRNA gene sequence. Representative isolates from each plant species were screened based on the kinetics of growth on fruit juices. A protocol for processing and storage of red and green smoothies (RS and GS) was set up, which included fermentation by selected lactic acid bacteria starters and exo-polysaccharide producing strains. Starters grew and remained viable at ca. 9.0 log cfu g(-1) during 30 days of storage at 4 °C. No contaminating Enterobacteriaceae and yeast were found throughout storage. Values of soluble solids, total titratable acidity and viscosity distinguished started RS and GS compared to spontaneously (unstarted) fermented smoothies. Color difference dE∗(ab) and browning index were positively affected by lactic acid fermentation. Consumption of carbohydrates by lactic acid bacteria was limited as well as it was the lactic fermentation. Consumption of malic acid was evident throughout storage. Polyphenolic compounds and, especially, ascorbic acid were better preserved in started RS and GS compared to unstarted samples. This reflected on the free radical scavenging activity. A statistical correlation was only found between the level of ascorbic acid and free radical scavenging activity. As shown by a first-order equation, the rate of degradation of ascorbic acid through storage were found to be higher in the unstarted compared to started RS and GS. Fermentation by lactic acid bacteria clearly improved the sensory attributes of RS and GS.

Antifungal activity of Wickerhamomyces anomalus and Lactobacillus plantarum during sourdough fermentation: identification of novel compounds and long-term effect during storage of wheat bread.

This study aimed at investigating the antifungal activity of Wickerhamomyces anomalus and sourdough lactic acid bacteria to extend the shelf life of wheat flour bread. The antifungal activity was assayed by agar diffusion, growth rate inhibition, and conidial germination assays, using Penicillium roqueforti DPPMAF1 as the indicator fungus. Sourdough fermented by Lactobacillus plantarum 1A7 (S1A7) and dough fermented by W. anomalus LCF1695 (D1695) were selected and characterized. The water/salt-soluble extract of S1A7 was partially purified, and several novel antifungal peptides, encrypted into sequences of Oryza sativa proteins, were identified. The water/salt-soluble extract of D1695 contained ethanol and, especially, ethyl acetate as inhibitory compounds. As shown by growth inhibition assays, both water/salt-soluble extracts had a large inhibitory spectrum, with some differences, toward the most common fungi isolated from bakeries. Bread making at a pilot plant was carried out with S1A7, D1695, or a sourdough started with a combination of both strains (S1A7-1695). Slices of the bread manufactured with S1A7-1695 did not show contamination by fungi until 28 days of storage in polyethylene bags at room temperature, a level of protection comparable to that afforded by 0.3% (wt/wt) calcium propionate. The effect of sourdough fermentation with W. anomalus LCF1695 was also assessed based on rheology and sensory properties.

Utilization of African grains for sourdough bread making.

UNLABELLED: Acha and Iburu flours were singly subjected to sourdough fermentation with previously selected autochthonous starters. Sourdoughs were used (30%, wt/wt) as aroma carriers and acidifiers during short time fermentation with the addition of baker's yeast. Acha and Iburu sourdough breads were compared to wheat sourdough bread started with the same strains and to breads made with the same formula but using baker's yeast alone. During Acha and Iburu sourdough fermentations, starter lactic acid bacteria reached almost the same cell density found in wheat sourdoughs. Acidification was more intense. Iburu sourdough bread had the highest total titratable acidity, the lowest pH, and contained the highest levels of free amino acids and phytase activity. The values of in vitro protein digestibility did not differ between Acha sourdough and wheat sourdough breads, while Iburu sourdough bread showed a slightly lower value. Acha and Iburu sourdough breads showed lower specific volume and higher density with respect to wheat sourdough breads. Nevertheless, Acha and Iburu sourdough breads were preferred for hardness and resilience. As shown by sensory analysis, Acha and especially Iburu sourdough breads were appreciated for color, acid taste and flavor, and overall acceptability. PRACTICAL APPLICATION: This study was aimed at evaluating the technological and nutritional properties of the African cereals Acha and Iburu. Sourdough fermentation and the use of selected starters increased the nutritional and sensory qualities and the potential application for bakery industry.