Pasquale Filannino

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.

Metabolic responses of Lactobacillus plantarum strains during fermentation and storage of vegetable and fruit juices.

ABSTRACT Strains of Lactobacillus plantarum were grown and stored in cherry (ChJ), pineapple (PJ), carrot (CJ), and tomato (TJ) juices to mimic the chemical composition of the respective matrices. Wheat flour hydrolysate (WFH), whey milk (W), and MRS broth were also used as representatives of other ecosystems. The growth rates and cell densities of L. plantarum strains during fermentation (24 h at 30°C) and storage (21 days at 4°C) differed only in part, being mainly influenced by the matrix. ChJ and PJ were the most stressful juices for growth and survival. Overall, the growth in juices was negatively correlated with the initial concentration of malic acid and carbohydrates. The consumption of malic acid was noticeable for all juices, but mainly during fermentation and storage of ChJ. Decreases of branched-chain amino acids (BCAA)—with the concomitant increase of their respective branched alcohols—and His and increases of Glu and gamma-aminobutyric acid (GABA) were the main traits of the catabolism of free amino acids (FAA), which were mainly evident under less acidic conditions (CJ and TJ). The increase of Tyr was found only during storage of ChJ. Some aldehydes (e.g., 3-methyl-butanal) were reduced to the corresponding alcohols (e.g., 3-methyl-1-butanol). After both fermentation and storage, acetic acid increased in all fermented juices, which implied the activation of the acetate kinase route. Diacetyl was the ketone found at the highest level, and butyric acid increased in almost all fermented juices. Data were processed through multidimensional statistical analyses. Except for CJ, the juices (mainly ChJ) seemed to induce specific metabolic traits, which differed in part among the strains. This study provided more in-depth knowledge on the metabolic mechanisms of growth and maintenance of L. plantarum in vegetable and fruit habitats, which also provided helpful information to select the most suitable starters for fermentation of targeted matrices.

Lactic acid fermentation as a tool to enhance the functional features of Echinacea spp

BackgroundExtracts and products (roots and/or aerial parts) from Echinacea ssp. represent a profitable market sector for herbal medicines thanks to different functional features. Alkamides and polyacetylenes, phenols like caffeic acid and its derivatives, polysaccharides and glycoproteins are the main bioactive compounds of Echinacea spp. This study aimed at investigating the capacity of selected lactic acid bacteria to enhance the antimicrobial, antioxidant and immune-modulatory features of E. purpurea with the prospect of its application as functional food, dietary supplement or pharmaceutical preparation.ResultsEchinacea purpurea suspension (5%, wt/vol) in distilled water, containing 0.4% (wt/vol) yeast extract, was fermented with Lactobacillus plantarum POM1, 1MR20 or C2, previously selected from plant materials. Chemically acidified suspension, without bacterial inoculum, was used as the control to investigate functional features. Echinacea suspension fermented with Lb. plantarum C2 exhibited a marked antimicrobial activity towards Gram-positive and -negative bacteria. Compared to control, the water-soluble extract from Echinacea suspension fermented with Lactobacillus plantarum 1MR20 showed twice time higher radical scavenging activity on DPPH. Almost the same was found for the inhibition of oleic acid peroxidation. The methanol extract from Echinacea suspension had inherent antioxidant features but the activity of extract from the sample fermented with strain 1MR20 was the highest. The antioxidant activities were confirmed on Balb 3T3 mouse fibroblasts. Lactobacillus plantarum C2 and 1MR20 were used in association to ferment Echinacea suspension, and the water-soluble extract was subjected to ultra-filtration and purification through RP-FPLC. The antioxidant activity was distributed in a large number of fractions and proportional to the peptide concentration. The antimicrobial activity was detected only in one fraction, further subjected to nano-LC-ESI-MS/MS. A mixture of eight peptides was identified, corresponding to fragments of plantaricins PlnH or PlnG. Treatments with fermented Echinacea suspension exerted immune-modulatory effects on Caco-2 cells. The fermentation with Lb. plantarum 1MR20 or with the association between strains C2 and 1MR20 had the highest effect on the expression of TNF-α gene.ConclusionsE. purpurea subjected to lactic acid fermentation could be suitable for novel applications as functional food dietary supplements or pharmaceutical preparations.

Hydroxycinnamic Acids Used as External Acceptors of Electrons: an Energetic Advantage for Strictly Heterofermentative Lactic Acid Bacteria

ABSTRACT The metabolism of hydroxycinnamic acids by strictly heterofermentative lactic acid bacteria (19 strains) was investigated as a potential alternative energy route. Lactobacillus curvatus PE5 was the most tolerant to hydroxycinnamic acids, followed by strains of Weissella spp., Lactobacillus brevis, Lactobacillus fermentum, and Leuconostoc mesenteroides, for which the MIC values were the same. The highest sensitivity was found for Lactobacillus rossiae strains. During growth in MRS broth, lactic acid bacteria reduced caffeic, p-coumaric, and ferulic acids into dihydrocaffeic, phloretic, and dihydroferulic acids, respectively, or decarboxylated hydroxycinnamic acids into the corresponding vinyl derivatives and then reduced the latter compounds to ethyl compounds. Reductase activities mainly emerged, and the activities of selected strains were further investigated in chemically defined basal medium (CDM) under anaerobic conditions. The end products of carbon metabolism were quantified, as were the levels of intracellular ATP and the NAD+/NADH ratio. Electron and carbon balances and theoretical ATP/glucose yields were also estimated. When CDM was supplemented with hydroxycinnamic acids, the synthesis of ethanol decreased and the concentration of acetic acid increased. The levels of these metabolites reflected on the alcohol dehydrogenase and acetate kinase activities. Overall, some biochemical traits distinguished the common metabolism of strictly heterofermentative strains: main reductase activity toward hydroxycinnamic acids, a shift from alcohol dehydrogenase to acetate kinase activities, an increase in the NAD+/NADH ratio, and the accumulation of supplementary intracellular ATP. Taken together, the above-described metabolic responses suggest that strictly heterofermentative lactic acid bacteria mainly use hydroxycinnamic acids as external acceptors of electrons.

Quorum-Sensing Regulation of Constitutive Plantaricin by Lactobacillus plantarum Strains under a Model System for Vegetables and Fruits

ABSTRACT 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.

Metabolic and functional paths of lactic acid bacteria in plant foods: get out of the labyrinth

Even though lactic acid bacteria are only a small part of the plant autochthonous microbiota, they represent the most important microbes having the capability to promote significant changes in the health-promoting properties of plant foods. Owing to the variety of plant chemical components and the possible pathways for bioconversion, plant fermentation is like a metabolic labyrinth undertaken by bacteria. The winding metabolic pathways involve several secondary plant metabolites (e.g. phenolics). The success of these paths is connected to the adaptive growth and survival of lactic acid bacteria. A panel of various interacting omics approaches unraveled the specific traits of lactic acid bacteria to adapt to plants, which allow the optimal design of fermentation strategies for targeted raw matrices.

Transcriptional reprogramming and phenotypic switching associated with the adaptation of Lactobacillus plantarum C2 to plant niches.

Lactobacillus plantarum has been isolated from a large variety of ecological niches, thus highlighting its remarkable environmental adaptability as a generalist. Plant fermentation conditions markedly affect the functional features of L. plantarum strains. We investigated the plant niche-specific traits of L. plantarum through whole-transcriptome and phenotypic microarray profiles. Carrot (CJ) and pineapple (PJ) juices were chosen as model systems, and MRS broth was used as a control. A set of 3,122 genes was expressed, and 21 to 31% of genes were differentially expressed depending on the plant niche and cell physiological state. L. plantarum C2 seemed to specifically respond to plant media conditions. When L. plantarum was cultured in CJ, useful pathways were activated, which were aimed to sense the environment, save energy and adopt alternative routes for NAD+ regeneration. In PJ the acidic environment caused a transcriptional switching, which was network-linked to an acid tolerance response involving carbohydrate flow, amino acid and protein metabolism, pH homeostasis and membrane fluidity. The most prominent phenotypic dissimilarities observed in cells grown in CJ and PJ were related to carbon and nitrogen metabolism, respectively. Summarising, a snapshot of a carrot and pineapple sensing and adaptive regulation model for L. plantarum C2 was proposed.

How to improve the gluten-free diet: The state of the art from a food science perspective

The celiac disease is the most common food intolerance and its prevalence is increasing. Consequently, use of gluten-free diet has expanded, notwithstanding consumption as therapy for other gluten-related disorders or by wellbeing people without any medical prescription. Even the therapeutic efficiency has undoubtedly proven, several drawbacks mainly regarding the compliance, nutritional deficits and related diseases, and the alteration of the intestinal microbiota have described in the literature. Food science has been considered as one of the primary area of intervention to limit or eliminate such drawbacks. Efforts have approached shelf life, rheology and palatability aspects but more recently have mainly focused to improve the nutritional features of the gluten-free diet, and to propose dietary alternatives. The sourdough fermentation has shown the most promising results, also including a biotechnology strategy that has allowed the complete degradation of gluten prior to consumption.

Lactic acid fermentation drives the optimal volatile flavor-aroma profile of pomegranate juice

Pomegranate juice (PJ) fermented with Lactobacillus plantarum C2, POM1, and LP09, unstarted-PJ, and raw-PJ were characterized for the profile of the volatile components (VOC) by PT-GC-MS. Lactic acid fermentation through selected strains enhanced the flavor profile of PJ. Concentrations of desired compounds (e.g., alcohols, ketones, and terpenes) were positively affected, whereas those of non-desired aldehydes decreased. Unstarted-PJ mainly differentiated from fermented PJs for the highest levels of aldehydes and sulfur compounds, and in lesser extent of furans, whereas alcohols, ketones, and alkenes followed by terpenes and benzene derivatives mainly differentiated fermented PJs. As expected, the lowest level of VOC was found in raw-PJ. VOC profile reflected on the sensory features of fermented PJs, unstarted-PJ, and raw-PJ, which were evaluated using a consensus modified flavor profile based on 13 attributes. Fermented PJs were mainly discriminated by the higher intensity of floral, fruity and anise notes than the controls.

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 bakers practices. Besides, this review would provide a critical view of this topic in order to avoid the speculation that in this field unavoidably arise.