Effect of Air Classification and Enzymatic and Microbial Bioprocessing on Defatted Durum Wheat Germ: Characterization and Use as Bread Ingredient.

Its high dietary fiber and protein contents and nutritional quality make defatted wheat germ (DWG) a valuable cereal by-product, yet its negative impact on food structure limits its use as a food ingredient. In this research, DWG underwent air classification, which identified two fractions with high fiber (HF) and low fiber/high protein (LF) contents, and a bioprocessing protocol, involving treatment with xylanase and fermentation with selected lactic acid bacterial strains. The degree of proteolysis was evaluated through electrophoretic and chromatographic techniques, revealing differences among fractions and bioprocessing options. Fermentation led to a significant increase in free amino acids (up to 6 g/kg), further enhanced by the combination with xylanase. When HF was used as an ingredient in bread making, the fiber content of the resulting bread exceeded 3.6 g/100 g, thus reaching the threshold required to make a "source of fiber" claim according to Regulation EC No.1924/2006. Meanwhile, all breads could be labeled a "source of protein" since up to 13% of the energy was provided by proteins. Overall, bioprocessed ingredients lowered the glycemic index (84 vs. 89) and increased protein digestibility (80 vs. 63%) compared to control breads. Technological and sensory analysis showed that the enzymatic treatment combined with fermentation also conferred a darker and more pleasant color to the bread crust, as well as better crumb porosity and elasticity.

Sourdough fermentation for the valorization of sorghum flour: Microbiota characterization and metabolome profiling.

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.

Role of tailored sourdough fermentation in the flavor of wholegrain-oat bread.

Sourdough technology has been known for its role in the improvement of texture, flavor, and quality of mainly wheat and rye-based breads for decades. However, little is reported about its use in the improvement of whole-grain oat bread, especially concerning flavor formation, which is one major consumer drivers. This study investigated the effects of sourdough obtained by different lactic acid bacteria and yeast starters consortia on the texture and flavor of 100% oat bread. Four different consortia were selected to obtain four oat sourdoughs, which were analyzed to assess the main features due to the different starter fermentation metabolism. Sourdoughs were added to breads as 30% dough weight. Bread quality was technologically monitored via hardness and volume measurements. Sourdough breads were softer and had higher specific volume. The sensory profile of sourdoughs and breads was assessed by a trained panel in sensory laboratory conditions, and the volatile profile was analyzed by HS-SPME-GC-MS. Sourdoughs were rated with higher intensities than untreated control for most of attributes, especially concerning sour aroma and flavor attributes. Sourdough breads were rated with higher intensities than control bread for sour vinegar flavor and total odor intensity, in addition they had richer volatile profile. Our results confirmed that sourdough addition can lead to an enhanced flavor, moreover, it demonstrated that the use of different consortia of lactic acid bacteria and yeast strains leads to the improvement of texture and altered sensory profile of whole-oat bread.

Comparison of Milk Kefirs Obtained from Cow's, Ewe's and Goat's Milk: Antioxidant Role of Microbial-Derived Exopolysaccharides.

Different types of milk are used in the production of milk kefir, but little information is available on the release of potentially antioxidant exopolysaccharides (EPS). The aim of this study was to investigate whether the microbial dynamics and EPS release are dependent on the milk substrate. In our study, the inoculated microbial consortium was driven differently by each type of milk (cow, ewe, and goat). This was evident in the sugar consumption, organic acid production, free amino release, and EPS production. The amount and the composition of the secreted EPS varied depending on the milk type, with implications for the structure and functional properties of the EPS. The low EPS yield in ewe's milk was associated with a higher lactic acid production and thus with the use of carbon sources oriented towards energy production. Depending on the milk used as substrate, the EPS showed different monosaccharide and FT-IR profiles, microstructures, and surface morphologies. These differences affected the antiradical properties and reducing power of the EPS. In particular, EPS extracted from cow's milk had a higher antioxidant activity than other milk types, and the antioxidant activity was concentration dependent.

Up-Cycling Grape Pomace through Sourdough Fermentation: Characterization of Phenolic Compounds, Antioxidant Activity, and Anti-Inflammatory Potential.

Despite its appealing composition, because it is rich in fibers and polyphenols, grape pomace, the major by-product of the wine industry, is still discarded or used for feed. This study aimed at exploiting grape pomace functional potential through fermentation with lactic acid bacteria (LAB). A systematic approach, including the progressively optimization of the grape pomace substrate, was used, evaluating pomace percentage, pH, and supplementation of nitrogen and carbon sources. When grape pomace was used at 10%, especially without pH correction, LAB cell viability decreased up to 2 log cycles. Hence, the percentage was lowered to 5 or 2.5% and supplementations with carbon and nitrogen sources, which are crucial for LAB metabolism, were considered aiming at obtaining a proper fermentation of the substrate. The optimization of the substrate enabled the comparison of strains performances and allowed the selection of the best performing strain (Lactiplantibacillus plantarum T0A10). A sourdough, containing 5% of grape pomace and fermented with the selected strain, showed high antioxidant activity on DPPH and ABTS radicals and anti-inflammatory potential on Caco2 cells. The anthocyanins profile of the grape pomace sourdough was also characterized, showing qualitative and quantitative differences before and after fermentation. Overall, the grape pomace sourdough showed promising applications as a functional ingredient in bread making.

Sourdough "Biga" Fermentation Improves the Digestibility of Pizza Pinsa Romana: An Investigation through a Simulated Static In Vitro Model.

Baked goods manufacturing parameters and fermentation conditions interfere with the nutrients content and affect their gastrointestinal fate. Pinsa Romana is a type of pizza that, recently, has been commercially rediscovered and that needed elucidation from a nutritional and digestibility perspective. In this study, six types of Pinsa Romana (five made with indirect method and one produced with straight dough technology) were characterized for their biochemical and nutritional features. Several variables like indirect (biga) Pinsa Romana production process, fermentation time and use of sourdough were investigated. The Pinsa Romana made with biga including sourdough and fermented for 48 h at 16 °C ((PR_48(SD)) resulted in the lowest predicted glycemic index, in the highest content of total peptides, total and individual free amino acids and gamma-amino butyric acid (GABA), and in the best protein quality indexes (protein efficiency ratio and nutritional index). The static in vitro digestion showed that the digesta from PR_48(SD) confirmed a reduced in vitro glycemic response after intake, and it showed a lower bioavailability of hydrophilic peptides. Furthermore, the inclusion of sourdough in biga enhanced the bioavailability of protein-related end-products including human health promoting compounds such as essential amino acids.

Use of Selected Lactic Acid Bacteria and Carob Flour for the Production of a High-Fibre and "Clean Label" Plant-Based Yogurt-like Product.

Carob, an underutilized crop with several ecological and economic advantages, was traditionally used as animal feed and excluded from the human diet. Yet, nowadays, its beneficial effects on health are making it an interesting candidate as a food ingredient. In this study, a carob-based yogurt-like product was designed and fermented with six lactic acid bacteria strains, whose performances after fermentation and during shelf life were assessed through microbial and biochemical characterization. The strains showed different aptitudes to ferment the rice-carob matrix. Particularly, Lactiplantibacillus plantarum T6B10 was among the strains with the lowest latency phase and highest acidification at the end of fermentation. T6B10 also showed discrete proteolysis during storage, so free amino acids were up to 3-fold higher compared to the beverages fermented with the other strains. Overall, fermentation resulted in the inhibition of spoilage microorganisms, while an increase in yeasts was found in the chemically acidified control. The yogurt-like product was characterized by high-fiber and low-fat content; moreover, compared to the control, fermentation decreased the predicted glycemic index (-9%) and improved the sensory acceptability. Thus, this work demonstrated that the combination of carob flour and fermentation with selected lactic acid bacteria strains represents a sustainable and effective option to obtain safe and nutritious yogurt-like products.

Bioprocessing to Preserve and Improve Microalgae Nutritional and Functional Potential: Novel Insight and Perspectives.

Microalgae are aquatic unicellular microorganisms and, although various species are approved for human consumption, Arthrospira and Chlorella are the most widespread. Several nutritional and functional properties have been bestowed to microalgae principal micro- and macro-nutrients, with antioxidant, immunomodulatory and anticancer being the most common. The many references to their potential as a food of the future is mainly ascribed to the high protein and essential amino acid content, but they are also a source of pigments, lipids, sterols, polysaccharides, vitamins, and phenolic compounds with positive effects on human health. Nevertheless, microalgae use is often hindered by unpleasant color and flavor and several strategies have been sought to minimize such challenges. This review provides an overview of the strategies so far proposed and the main nutritional and functional characteristic of microalgae and the foods made thereof. Processing treatments have been used to enrich microalgae-derived substrates in compounds with antioxidant, antimicrobial, and anti-hypertensive properties. Extraction, microencapsulation, enzymatic treatments, and fermentation are the most common, each with their own pros and cons. Yet, for microalgae to be the food of the future, more effort should be put into finding the right pre-treatments that can allow the use of the whole biomass and be cost-effective while bringing about features that go beyond the mere increase of proteins.

Probiotic Potential and Safety Assessment of Type Strains of Weissella and Periweissella Species.

Although numerous strains belonging to the Weissella genus have been described in the last decades for their probiotic and biotechnological potential, others are known to be opportunistic pathogens of humans and animals. Here, we investigated the probiotic potential of two Weissella and four Periweissella type strains belonging to the species Weissella diestrammenae, Weissella uvarum, Periweissella beninensis, Periweissella fabalis, Periweissella fabaria, and Periweissella ghanensis by genomic and phenotypic analyses, and performed a safety assessment of these strains. Based on the results of the survival to simulated gastrointestinal transit, autoaggregation and hydrophobicity characteristics, as well as adhesion to Caco-2 cells, we showed that the P. beninensis, P. fabalis, P. fabaria, P. ghanensis, and W. uvarum type strains exhibited a high probiotic potential. The safety assessment, based on the genomic analysis, performed by searching for virulence and antibiotic resistance genes, as well as on the phenotypic evaluation, by testing hemolytic activity and antibiotic susceptibility, allowed us to identify the P. beninensis type strain as a safe potential probiotic microorganism. IMPORTANCE A comprehensive analysis of safety and functional features of six Weissella and Periweissella type strains was performed. Our data demonstrated the probiotic potential of these species, indicating the P. beninensis type strain as the best candidate based on its potential probiotic features and the safety assessment. The presence of different antimicrobial resistance profiles in the analyzed strains highlighted the need to establish cutoff values to perform a standardized safety evaluation of these species, which, in our opinion, should be mandatory on a strain-specific basis.

Design of a Plant-Based Yogurt-Like Product Fortified with Hemp Flour: Formulation and Characterization.

Plant-based milk alternatives have gained massive popularity among consumers because of their sustainable production compared to bovine milk and because of meeting the nutritional requests of consumers affected by cow milk allergies and lactose intolerance. In this work, hemp flour, in a blend with rice flour, was used to design a novel lactose- and gluten-free yogurt-like (YL) product with suitable nutritional, functional, and sensory features. The growth and the acidification of three different lactic acid bacteria strains were monitored to better set up the biotechnological protocol for making the YL product. Hemp flour conferred the high fiber (circa 2.6 g/100 g), protein (circa 4 g/100 g), and mineral contents of the YL product, while fermentation by selected lactic acid bacteria increased the antioxidant properties (+8%) and the soluble fiber (+0.3 g/100 g), decreasing the predicted glycemic index (-10%). As demonstrated by the sensory analysis, the biotechnological process decreased the earthy flavor (typical of raw hemp flour) and increased the acidic and creamy sensory perceptions. Supplementation with natural clean-label vanilla powder and agave syrup was proposed to further decrease the astringent and bitter flavors. The evaluation of the starter survival and biochemical properties of the product under refrigerated conditions suggests an estimated shelf-life of 30 days. This work demonstrated that hemp flour might be used as a nutritional improver, while fermentation with a selected starter represents a sustainable and effective option for exploiting its potential.

Modeling of Growth and Organic Acid Kinetics and Evolution of the Protein Profile and Amino Acid Content during Lactiplantibacillus plantarum ITM21B Fermentation in Liquid Sourdough.

The application of mathematical modeling to study and characterize lactic acid bacterial strains with pro-technological and functional features has gained attention in recent years to solve the problems relevant to the variabilities of the fermentation processes of sourdough. Since the key factors contributing to the sourdough quality are relevant to the starter strain growth and its metabolic activity, in this study, the cardinal growth parameters for pH, temperature (T), water activity (aw), and undissociated lactic acid of the sourdough strain Lactiplantibacillus plantarum ITM21B, were determined. The strain growth, pH, organic acids (lactic, acetic, phenyllactic, and hydroxy-phenyllactic), total free amino acids, and proteins were monitored during fermentation of a liquid sourdough based on wheat flour and gluten (Bio21B) after changing the starting T, pH, and inoculum load. Results demonstrated that the different fermentation conditions affected the strain growth and metabolite pattern. The organic acid production and growth performance were modeled in Bio21B, and the resulting predictive model allowed us to simulate in silico the strain performances in liquid sourdough under different scenarios. This mathematical predictive approach can be useful to optimize the fermentation conditions needed to obtain the suitable nutritional and technological characteristics of the L. plantarum ITM21B liquid sourdough.

The Potential of Food By-Products: Bioprocessing, Bioactive Compounds Extraction and Functional Ingredients Utilization.

Achieving sustainability in the agro-food sector can only be possible with the valorization of food industry waste and side streams, products with an extremely high intrinsic value but often discarded because they are unfit for further processing that meets consumer expectations [...].

Use of Selected Lactic Acid Bacteria for the Fermentation of Legume-Based Water Extracts.

In this study, the effect of selected Lactobacillus acidophilus ATCC 4356, Limosilactobacillus fermentum DSM 20052, and Lacticaseibacillus paracasei subsp. paracasei DSM 20312 strains on the sensory characteristics, and protein and amino acid content of fermented water extracts derived from lupin, pea, and bean grains is reported. Even though all strains were able to grow over 7 log cfu mL-1 and to decrease pH in the range of -0.52 to -1.25 within 24 h, the release of an unpleasant ferric-sulfurous off-odor from the fermented bean water extract prohibited further characterization. Lupin and pea grain-based beverages underwent an in-depth sensory evaluation using a simplified check-all-that-apply (CATA) method, finding new and appreciable sensory notes such as cooked ham, almonds, and sandalwood. Fermented lupin water extract showed higher total protein content (on average, 0.93 mg mL-1) in comparison to that of pea grains (on average, 0.08 mg mL-1), and a free amino acid content (on average, 3.9 mg mL-1) close to that of cow milk. The concentrations of these nutrients decreased during refrigerated storage, when the lactic acid bacteria load was always higher than 7 log cfu mL-1. The results of this study indicated that lactic fermentation improves the sensory characteristics of these innovative legume-based beverages, which sustained high loads of viable lactobacilli up to the end of cold storage.

Exploitation of wasted bread as substrate for polyhydroxyalkanoates production through the use of Haloferax mediterranei and seawater.

The use of the halophile microorganism Haloferax mediterranei, able to synthesize poly(hydroxybutyrate-hydroxyvalerate) (PHBV), is considered as a promising tool for the industrial production of bioplastic through bioprocessing. A consistent supplementation of the growth substrate in carbohydrates and minerals is overall necessary to allow its PHBV production. In this work, wasted bread was used as substrate for bioplastic production by microbial fermentation. Instead of the consistent and expensive minerals supplement required for Hfx. mediterranei DSM1411 growth, microfiltered seawater was added to the wasted bread-derived substrate. The suitable ratio of wasted bread homogenate and seawater, corresponding to 40:60, was selected. The addition of proteases and amylase to the bread homogenate promoted the microbial growth but it did not correspond to the increase of bioplastic production by the microorganism, that reach, under the experimental conditions, 1.53 g/L. An extraction procedure of the PHBV from cells, based on repeated washing with water, followed or not by a purification through ethanol precipitation, was applied instead of the conventional extraction with chloroform. Yield of PHBV obtained using the different extraction methods were 21.6 ± 3.6 (standard extraction/purification procedure with CHCl3:H2O mixture), 24.8 ± 3.0 (water-based extraction), and 19.8 ± 3.3 mg PHAs/g of wasted bread (water-based extraction followed by ethanol purification). Slightly higher hydroxyvalerate content (12.95 vs 10.78%, w/w) was found in PHBV obtained through the water-based extraction compared to the conventional one, moreover, the former was characterized by purity of 100% (w/w). Results demonstrated the suitability of wasted bread, supplemented with seawater, to be used as substrate for bioplastic production through fermentation. Results moreover demonstrated that a solvent-free extraction, exclusively based on osmotic shock, could be used to recover the bioplastic from cells.

Interaction between Fish Skin Gelatin and Pea Protein at Air-Water Interface after Ultrasound Treatment.

The interaction between fish skin gelatin (FG) and pea protein isolate (PPI) was investigated at the air-water interface (A-W) before and after a high intensity (275 W, 5 min) ultrasound treatment (US). We analyzed the properties of the single protein suspensions as well as an equal ratio of FG:PPI (MIX), in terms of ζ-potential, particle size, molecular weight, bulk viscosity and interfacial tension. The foaming properties were then evaluated by visual analysis and by Turbiscan Tower. Confocal laser scanning microscopy (CLSM) was employed to explore the role of the proteins on the microstructure of foams. The results showed that the ultrasound treatment slightly influenced physicochemical properties of the proteins, while in general, did not significantly affect their behavior both in bulk and at the air-water interface. In particular, PPI aggregate size was reduced (-48 nm) while their negative charges were increased (-1 mV) after the treatment. However, when the proteins were combined, higher molecular weight of aggregates, higher foam stability values (+14%) and lower interfacial tension (IFT) values (47.2 ± 0.2 mN/m) were obtained, leading us to assume that a weak interaction was developed between them.

Reuse of Wasted Bread as Soil Amendment: Bioprocessing, Effects on Alkaline Soil and Escarole (Cichorium endivia) Production.

In an era characterized by land degradation, climate change, and a growing population, ensuring high-yield productions with limited resources is of utmost importance. In this context, the use of novel soil amendments and the exploitation of plant growth-promoting microorganisms potential are considered promising tools for developing a more sustainable primary production. This study aimed at investigating the potential of bread, which represents a large portion of the global food waste, to be used as an organic soil amendment. A bioprocessed wasted bread, obtained by an enzymatic treatment coupled with fermentation, together with unprocessed wasted bread were used as amendments in a pot trial. An integrated analytical plan aimed at assessing (i) the modification of the physicochemical properties of a typical Mediterranean alkaline agricultural soil, and (ii) the plant growth-promoting effect on escarole (Cichorium endivia var. Cuartana), used as indicator crop, was carried out. Compared to the unamended soils, the use of biomasses raised the soil organic carbon content (up to 37%) and total nitrogen content (up to 40%). Moreover, the lower pH and the higher organic acid content, especially in bioprocessed wasted bread, determined a major availability of Mn, Fe, and Cu in amended soils. The escaroles from pots amended with raw and bioprocessed bread had a number of leaves, 1.7- and 1.4-fold higher than plants cultivated on unamended pots, respectively, showing no apparent phytotoxicity and thus confirming the possible re-utilization of such residual biomasses as agriculture amendments.

Biosynthesis of γ-aminobutyric acid by lactic acid bacteria in surplus bread and its use in bread making.

AIMS: The aim of this study was to investigate the effectiveness of bread as substrate for γ-aminobutyric acid (GABA) biosynthesis, establishing a valorization strategy for surplus bread, repurposing it within the food chain. METHODS AND RESULTS: Surplus bread was fermented by lactic acid bacteria (LAB) to produce GABA. Pediococcus pentosaceus F01, Levilactobacillus brevis MRS4, Lactiplantibacillus plantarum H64 and C48 were selected among 33 LAB strains for the ability to synthesize GABA. Four fermentation experiments were set up using surplus bread as such, added of amylolytic and proteolytic enzymes, modifying the pH or mixed with wheat bran. Enzyme-treated slurries led to the release of glucose (up to 20 mg g-1 ) and free amino acid, whereas the addition of wheat bran (30% of bread weight) yielded the highest GABA content (circa 800 mg kg-1 of dry weight) and was the most suitable substrate for LAB growth. The selected slurry was ultimately used as an ingredient in bread making causing an increase in free amino acids. CONCLUSIONS: Besides the high GABA concentration (148 mg kg-1 dough), the experimental bread developed in this study was characterized by good nutritional properties, highlighting the efficacy of tailored bioprocessing technologies as means to mitigate food wastage. SIGNIFICANCE AND IMPACT OF STUDY: Our results represent a proof of concept of effective strategies to repurpose food industry side streams.

Lactic Acid Bacteria Fermentation and Endopeptidase Treatment Improve the Functional and Nutritional Features of Arthrospira platensis.

This study aimed at investigating the effect of fermentation and enzymatic treatment on the degree of proteolysis of wet (WB), dried at low temperature (DB), and freeze-dried Spirulina (LB) proteins that affect the nutritional (e.g., amino acid content and profiles, and protein digestibility) and functional (e.g., antioxidant and antimicrobial activities) properties. The desiccation treatments influenced the unprocessed Spirulina characteristics because, compared with that in WB, peptides and free amino acids content was 73% lower in DB and 34% higher in LB. An integrated approach, including chromatographic and electrophoresis analyses, was used to evaluate the effect of the different bioprocessing options on protein profiles, release of peptides and amino acids, and the overall protein digestibility. Compared with the application of fermentation with the selected Lactiplantibacillus plantarum T0A10, the treatment with the endopeptidase Alcalase®, alone or combined, determined the most intense proteolysis. Moreover, the treatment with Alcalase® of LB allowed the release of potentially bioactive compounds that are able to inhibit Penicillium roqueforti growth, whereas the combination of fermentation with L. plantarum T0A10 and Alcalase® treatment increased Spirulina antioxidant properties, as determined by the scavenging activity toward ABTS radical (up to 60%) and antimicrobial activity against food pathogen Escherichia coli.