Conjugation of gluten hydrolysates with glucosamine at mild temperatures enhances antioxidant and antimicrobial properties

Gluten represents one of the principal by-products of the wheat starch industry. Peptides obtained by wheat hydrolysis can be used for specific functional and biological activities, albeit at relatively low yields. Although the Maillard reaction (glycation) is widely used to increase functionality of proteins, its main disadvantage is the production of undesirable compounds due to high processing temperature. In this research, functional and biologically active glycopeptides were obtained from gluten. Alcalase or Flavourzyme proteases were used to hydrolyse gluten protein, and the resulting peptides were conjugated with glucosamine by enzymatic glycosylation, using transglutaminase, or through glycation. Both reactions were performed at mild temperatures (25 or 37°C). The formation of glycopeptides depended mostly on the glycation process, as demonstrated by MALDI-TOF-MS. The bioactivities of the conjugated hydrolysates were compared to the native hydrolysates. Although a reduction in the anti-ACE activity was detected, improved DPPH scavenging activity and enhanced antimicrobial activity against Escherichia coli were observed in the glycated Alcalase-derived hydrolysates and in the glycated Flavourzyme-derived hydrolysates, respectively. This study showed that mild conditions are an alternate approach to the traditional Maillard process conducted at elevated temperatures in creating conjugated gluten hydrolysates with enhanced bioactivities. © 2014 Elsevier Ltd

Dynamic Stresses of Lactic Acid Bacteria Associated to Fermentation Processes

Despite their negligible mass the microbial agents, starters and non starters, play a profound role in the characterization of the fermented foods in terms of chemical and sensorial properties. In fact, fermented foods may be defined as foods processed through the activity of microorganisms. Fermentation processes take a special place in the evolution of human cuisine, by altering the taste experience of food products, as well as extending the storage period. In particular, foods fermented with lactic acid bacteria (LAB) have constituted an important part of human diet and of fermentation processes (involving various foods, including milk, meat, vegetables and fruits) since ancient times. They have played an essential role in the preservation of agricultural resources and in the improvement of nutritional and organoleptic properties of human foods and animal feed. Moreover, these organisms nowadays are increasingly used as health promoting probiotics, enzyme and metabolite factories and vaccine delivery vehicles

Primi risultati del progetto LIFE+ sulle analisi microbiologiche delle acque nel Parco dei Gessi dell’Emilia Romagna

Dal 2010 è in corso il Progetto Life + 08NAT/IT/000369 “Gypsum” 2, cofinanziato dall’Unione Europea, finalizzato alla tutela e gestione dei principali ambienti gessosi dell’Emilia Romagna. Nell’ambito dell’Azione A3 è previsto un monitoraggio pluriennale dei principali acquiferi carsici sotto l’aspetto chimico e microbiologico. Nel corso del primo anno sono state analizzate le acque carsiche su circa 50 punti di controllo (inghiottitoi, fiumi, torrenti in grotta, e risorgenti). In generale l’obiettivo di questa sperimentazione è quello di valutare l’impatto di sostanze di origine agricola o di altre forme di inquinamento, legate ad insediamenti o attività antropiche o fattori naturali, in acque di grotta. La sperimentazione è stata sviluppata tramite tecniche microbiologiche classiche e di biologia molecolare (PCR 16S rRNA e PCR-DGGE), finalizzate alla caratterizzazione delle popolazioni microbiche presenti nei diversi siti di prelievo e alla determinazione di loro eventuali variazioni e/o evoluzioni. I valori di carica microbica totale determinati oscillavano da un massimo di 3.32 ad un minimo di 0.18 log UFC/ ml e da un massimo di 2.26 fino a valori al di sotto del limite di determinazione (1 log UFC/ml) per quanto riguarda i coliformi totali e fecali. Le analisi genetiche hanno mostrato la presenza di numerosi specie batteriche (Agrobacterium tumefaciens, Pseudomonas spp., Rahnella aquatilis, Stenotrophomonas maltophilia, Pedobacter swuonensis, Enterobacter spp., Aeromonas hydrophila, Citrobacter, Klebsiella and Raoultella). I microrganismi identificati possono avere diverse origini, alcuni provengono dal terreno, altri possono essere comuni contaminanti delle acque ed altri avere un’origine antropica (batteri fecali). Fino a questo step del progetto, l’analisi PCR-DGGE ha evidenziato le evoluzioni ecologiche, in termine di popolazioni microbiche, presenti tra i diversi campioni e i diversi siti di campionamento all’interno di una stessa grotta.The Project Life + 08NAT/IT/000369 “Gypsum” 2, co-financed by the European Union, has started in the spring of 2010. This project aims to protect and manage the main karst caves and sites of Emilia-Romagna region. The A3 action provides a periodic monitoring of the main karst aquifers in terms of chemistry and microbiology. During the first year and a half, karst waters of 50 control points were analysed (sinking streams, rivers and streams in caves, and resurgences). The objective of this study is to evaluate the impact, in the waters of the cave, of agricultural substances or other forms of pollution or settlements related to human activities or natural factors. The experiment was developed using traditional microbiology techniques and molecular biology techniques (PCR and 16S rRNA PCR-DGGE), focused on the characterization of microbial populations in the different sampling sites and determination of their variations and/or changes. The total microbial concentration ranged from a maxiimum of 3.32 or 2.26 to values below the limit of detection (1 log CFU/ml) for total and faecal colifroms, respectively. The genetic analysis showed the presence of numerous bacterial species (Agrobacterium tumefaciens, Pseudomonas spp., Rahnella aquatilis, Stenotrophomonas maltophilia, Pedobacter swuonensis, Enterobacter spp., Aeromonas hydrophila, Citrobacter, Klebsiella and Raoultella). The organisms identified have different origins, some come from the ground, others are common water contaminants and others derive from human activities (faecal bacteria). Up to now, PCR-DGGE revealed the ecological changes, in terms of microbial populations present in the samples, and different sampling sites within the same cave

Use of Yarrowia lipolytica to Obtain Fish Waste Functional Hydrolysates Rich in Flavoring Compounds

Fishery processing industries generate large amounts of by-products. These by-products come from fish heads, skin, bones, thorns, and viscera. The disposal of these wastes represents an increasing environmental and health problem. Nowadays, there is a growing interest in how to utilize fish materials that are not used for human consumption. Among the different solutions proposed, the use of proteolytic and lipolytic microorganisms represents a green solution for waste valorization. In this work, first we screened several conventional and non-conventioal microorganisms for their proteolytic and lipolytic functions. Then, the most promising strains (Yarrowia lipolytica YL2, Y. lipolytica YL4, Bacillus amyloliquefaciens B5M and B. subtilis B5C) were tested on a fish waste-based solution. After 72 h incubation at room temperature, the supernatants obtained using the strains of Y. lipolytica showed the highest degree of hydrolysis (10.03 and 11.80%, respectively, for YL2 and YL4), the strongest antioxidant activity (86.4% in DPPH assay for YL2) and the highest formation of aldehydes (above 50% of the total volatile compounds detected). Hydrolysates of fish waste obtained with Y. lipolytica may be reused in feed and food formulations for their functional and flavoring characteristics

Effects of Sub-Lethal High Pressure Homogenization Treatment on the Adhesion Mechanisms and Stress Response Genes in Lactobacillus acidophilus 08

Cell surface hydrophobicity (CSH) and adhesion are very important phenotypical traits for probiotics that confer them a competitive advantage for the resilience in the human gastrointestinal tract. This study was aimed to understand the effects over time of a 50 MPa hyperbaric treatment on the surface properties of Lactobacillus acidophilus 08 including CSH, autoaggregation, and in vitro adhesion (mucin layer and Caco-2 cells). Moreover, a link between the hurdle applied and the expression of genes involved in the general stress response (groEL and clpP) and adhesion processes (efTu and slpA) was evaluated. High pressure homogenization (HPH) at 50 MPa significantly increased the CSH percentage (H%), autoaggregation and in vitro adhesion on mucin of L. acidophilus 08 cells compared with the untreated cells. Moreover, the hyperbaric hurdle induced an upregulation of the stress response genes groEL and ef-TU together with a down regulation of the clpP and S-layer slpA genes. Looking at the protein profile, HPHtreatment showed an increase in the number or intensity of protein bands at high and low molecular weights

Effects of Starter Cultures and Type of Casings on the Microbial Features and Volatile Profile of Fermented Sausages

In the literature, the effect of the type of casing on fermented sausages is quite unexplored, while several studies are focused on the impact of starter cultures. Therefore, this paper studied the effect of three commercial starter cultures and two casings (natural or collagen) on Italian fermented sausages. Physico-chemical parameters (aw, pH, weight loss), microbiota, aroma profile and sensory analysis were evaluated. Results showed that collagen casings promoted a higher reduction of pH and weight loss. Concerning the microbiota, samples with natural casing had higher counts of lactic acid bacteria, while yeast proliferation was promoted in those with collagen. Regardless of the starters and casings applied, levels of enterococci and Enterobacteriaceae were low (≤2 log CFU/g). The aroma profile was significantly affected by casing: despite the starter applied, the presence of collagen casing favoured acid accumulation (mainly acetate and butanoate) and reduction of ketones. Sensory analysis highlighted significant differences only for odour, colour intensity and sourness. The differences observed suggest that collagen casings may provide a greater availability of oxygen. Overall, casings rather than starter cultures impact the microbial and sensorial features of fermented sausages

Development and characterization of fermented soy beverages containing encapsulated or non-encapsulated vaginal probiotics

Human microbial niches such as the healthy vagina, are recently emerging as “unconventional” sources of candidate probiotics capable of preventing from different vaginal diseases. These microorganisms could be provided as oral preparations since they can reach the vaginal niche passing through the gastrointestinal tract. However, their use in food would be challenging. The aim of this work was to develop and characterize fermented soy beverages with encapsulated and non-encapsulated vaginal lactobacilli, namely Lactobacillus crispatus BC4 and Lactobacillus gasseri BC9, as future dietary strategies for vaginal dysbiosis. The viability of vaginal strains remained stable at 7 log CFU/mL of product during the entire 28 days of storage, despite the use of encapsulated or non-encapsulated bacteria. Samples containing encapsulated bacteria, especially E-BC4+BC9, showed higher Water Holding Capacity (62.29%), lactic acid content (1.43%), and a remarkable antagonistic activity against enteropathogens. Moreover, encapsulation protected the strains from simulated GIT conditions (>1 Log) but reduced the acceptability of the final products. Overall, strain BC4 and BC9, alone or in mix, demonstrated to be promising co-starter cultures providing a characteristic flavor (pleasant smell and taste) and aroma (lower hexanal, benzaldehyde and higher diacetyl, and 2,3-pentanedione, compared to control) to the fermented soy beverages

Eucalyptus essential oil as a natural food preservative: in vivo and in vitro antiyeast potential

In this study, the application of eucalyptus essential oil/vapour as beverages preservative is reported. The chemical composition of eucalyptus oil was determined by gas chromatography-mass spectrometry (GC-MS) and solid phase microextraction GC-MS (SPME/GC-MS) analyses. GC-MS revealed that the major constituents were 1,8-cineole (80.5%), limonene (6.5%), \u3b1-pinene (5%), and \u3b3-terpinene (2.9%) while SPME/GC-MS showed a relative reduction of 1,8-cineole (63.9%) and an increase of limonene (13.8%), \u3b1-pinene (8.87%), and \u3b3-terpinene (3.98%). Antimicrobial potential of essential oil was initially determined in vitro against 8 different food spoilage yeasts by disc diffusion, disc volatilization, and microdilution method. The activity of eucalyptus vapours was significantly higher than the eucalyptus oil. Minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC) varied from 0.56 to 4.50\u2009mg/mL and from 1.13 to 9\u2009mg/mL, respectively. Subsequently, the combined efficacy of essential oil and thermal treatment were used to evaluate the preservation of a mixed fruit juice in a time-dependent manner. These results suggest eucalyptus oil as a potent inhibitor of food spoilage yeasts not only in vitro but also in a real food system. Currently, this is the first report that uses eucalyptus essential oil for fruit juice preservation against food spoiling yeast

Essential Oils and Their Combination with Lactic Acid Bacteria and Bacteriocins to Improve the Safety and Shelf Life of Foods: A Review

The use of plant extracts (e.g., essential oils and their active compounds) represents an interesting alternative to chemical additives and preservatives applied to delay the alteration and oxidation of foods during their storage. Essential oils (EO) are nowadays considered valuable sources of food preservatives as they provide a healthier alternative to synthetic chemicals while serving the same purpose without affecting food quality parameters. The natural antimicrobial molecules found in medicinal plants represent a possible solution against drug-resistant bacteria, which represent a global health problem, especially for foodborne infections. Several solutions related to their application on food have been described, such as incorporation in active packaging or edible film and direct encapsulation. However, the use of bioactive concentrations of plant derivatives may negatively impact the sensorial characteristics of the final product, and to solve this problem, their application has been proposed in combination with other hurdles, including biocontrol agents. Biocontrol agents are microbial cultures capable of producing natural antimicrobials, including bacteriocins, organic acids, volatile organic compounds, and hydrolytic enzymes. The major effect of bacteriocins or bacteriocin-producing LAB (lactic acid bacteria) on food is obtained when their use is combined with other preservation methods. The combined use of EOs and biocontrol agents in fruit and vegetables, meat, and dairy products is becoming more and more important due to growing concerns about potentially dangerous and toxic synthetic additives. The combination of these two hurdles can improve the safety and shelf life (inactivation of spoilage or pathogenic microorganisms) of the final products while maintaining or stabilizing their sensory and nutritional quality. This review critically describes and collects the most updated works regarding the application of EOs in different food sectors and their combination with biocontrol agents and bacteriocins

Survival, growth, and biogenic amine production of Enterococcus faecium FC12 in response to extracts and essential oils of Rubus fruticosus and Juniperus oxycedrus

Enterococci are lactic acid bacteria (LAB) usually found as food contaminants in fermented products such as cheeses and fermented sausages. Due to their antibiotic resistance, the presence of virulence factors, and the ability to produce biogenic amines (BAs), the determination of these bacteria is crucial to assure food quality and safety. BAs production and consequent accumulation in foods can cause toxicological eects on human health. Plant phenolic compounds are promising alternatives to chemical preservatives and reflect consumers’ demand for “green” solutions. In this study, the antimicrobial eect of blackberry (Rubus fruticosus) leaves and prickly juniper (Juniperus oxycedrus) needles, both as phenolic extracts (PE) and essential oils (EO), were evaluated against Enterococcus faecium FC12, a known tyramine-producing strai