Glutathione release in extracellular form by S. cerevisiae strains

Glutathione (GSH, L-\uf067-glutamyl-L-cysteinyl-glycine) is the most abundant non-protein thiol compound widely present in living organisms, from prokaryotes to eukaryotes (Anderson 1998). It is synthesised intracellularly by the consecutive actions of \uf067-glutamylcysteine synthetase, feedback inhibited by GSH, and GSH synthetase. This tripeptide\u2019s very low redox potential gives it the properties of a cellular redox buffer (Udeh and Achremowicz 1997). In living tissues, GSH plays a pivotal role in bioreduction, protection against oxidative stress, xenobiotic and endogenous toxic metabolite detoxification, enzyme activity and sulphur and nitrogen metabolism (Penninckx 2002). These characteristics make this active tripeptide an important aid and/or support for the treatment of numerous diseases, such as HIV infections, liver cirrhosis, pancreatic inflammations and aging (Wu et al., 2004). In addition, GSH is of interest in the food additive industry and sports nutrition (Lomaestro and Malone 1995). Yeasts, in particular belonging to the genus Saccharomyces, are the most commonly used microorganisms on an industrial scale for GSH fermentative production; however GSH contents of the wild-type strains are usually variable (0.1 \u2013 1% dw) and always in intracellular form (Rollini and Manzoni 2006). The present research was aimed at obtaining GSH in extracellular form, released from cells, at high levels. Samples of S. cerevisiae (baker\u2019s yeast) from different suppliers were tested, together with reference strains belonging to international collections. Cells were comparatively treated employing physical and chemical procedures. The best result (2.9 g/l, 90% of produced GSH in extracellular form) was achieved at 24 h reaction, employing lyophilised cells from compressed baker\u2019s yeast. The possibility of obtaining GSH directly in extracellular form, skipping the downstream cell extraction step, represents an interesting opportunity of reducing GSH production cost and furthering the range of application and utilization of this molecule

Zymomonas mobilis in Bread Dough: Characterization of Dough Leavening Performance in Presence of Sucrose

Zymomonas mobilis, because of its fermentative metabolism, has potential food applications in the development of leavened baked goods consumable by people with adverse responses to Saccharomyces cerevisiae. Since Z. mobilis is not able to utilize maltose present in flour, the effect of sucrose addition (2.5 g/100 g flour) on bread dough leavening properties was studied. For comparison purposes, leavening performances of S. cerevisiae with and without sucrose were also investigated. Doughs leavened by Z. mobilis without sucrose addition showed the lowest height development (14.95 +/- 0.21 mm) and CO2 production (855 +/- 136 mL). When sucrose was added, fermentative performances of Z. mobilis significantly (p < 0.05) improved (+80% and +85% of gas production and retention, respectively), with a dough maximum height 2.6 times higher, results indicating that Z. mobilis with sucrose can be leavened in shorter time with respect to the sample without addition. S. cerevisiae did not benefit the sucrose addition in terms of CO2 production and retention, even if lag leavening time was significantly (p < 0.05) shorter (about the half) and time of porosity appearance significantly (p < 0.05) longer (about 26%) with respect to S. cerevisiae alone. Results demonstrate that in the presence of sucrose, Z. mobilis can efficiently leaven a bread dough, thus providing innovation possibilities in the area of yeast-free leavened products

Posidonia Oceanica And Green Tea Extract As Active Agents For Shelf Life Extension Of Fresh Cut Peach

Nowadays the reduction of food waste and the availability of food all over the world areas are priority keys for society. In this context, the shelf life extension of fresh products in a sustainable way may represent one of the main goals. The aim of the present research work was to extend the shelf life of fresh cut peach slices by using extracts of Posidonia oceanica (POS) and green tea (GT), and compared to the control. Total polyphenols content was assessed by Folin-Ciocalteu method and the antioxidant capacity by DPPH (2,2-Diphenyl-1-Picrylhydrazyl) assay. Antimicrobial activity of the two extracts was initially carried out in vitro by determining the Minimal Inhibitory Concentration (MIC) against Escherichia coli, Listeria innocua, Pseudomonas putida, Staphylococcus aureus, Saccharomyces cerevisiae, Aspergillus niger and Penicillium chrysogenum. The two extracts in vivo on peach slices by dipping were applied (only water for the control), than, slices were placed into low-density polyethylene (LDPE) bag and stored at 3 ± 1°C for up to 7 days. Microbiological analyses, color (L*,a*,b*), total soluble solid (TSS) and titratable acidity (TA) on peach slices were carried out every 3 days. POS showed the highest polyphenols content (615±45 mg Gallic Acid Equivalent/g – mg GAE/g) while GT 526 ± 28 mg GAE/g. The values of EC50, for POS, was 72.42 ± 22.90 mg/L, while for GT 3.80 ± 0.11 mg/L. Trials in vitro showed MIC values of above 2 g/L for POS and above 1 g/L in GT mainly against Gram positive bacteria. POS was also found to lead a growth delay in the case of fungi, approximately 60 h compared to the control. Results related to the Total Aerobic (TAC) and yeasts and moulds (Y&M) counts highlighted that peach slices dipped with POS maintained in the first 5 days of storage the microbiota at lower or analogous levels than those found at t0 (1.80 ± 0.09 log cfu/g respect 2.30 ± 0.11 log cfu/g and 1.80 ± 0.05 log cfu/g respect 1.70 ± 0.03 log cfu/g of the control, respectively for TAC and Y&M). Regarding pomological analysis, after seven days, lightness of the peach slices decreased 28% in POS, 24% in GT and 35% in control. During shelf life TSS increased 8% in the control, 2% in POS and remained unchanged in GT, after seven days. At the same time, TA decreased 30% in POS, 20% in GT treatment and 63% in the control. During the experiment TSS increased in all treatments, more rapidly in the control; meanwhile the TA decreased slowly in treated samples compared to the control. Concluding, Posidonia oceanica and green tea extracts were found able to delay the colour and pomological parameters decay, maintaining a good fruit quality during shelf life. Data already showed evidence a better performance of the Posidonia extract, limiting the growth of microorganisms responsible for fresh fruit deterioration

INTERACTIVE POLYPHENOLS-BASED BIOPACKAGING FOR FOOD PRESERVATION: AN IN VITRO STUDY

Producing green, sustainable and renewable materials is one of the major challenge nowadays in the food-packaging sector. In this context, the aim of our study is to develop an active cellulose-based packaging, cellulose in fact is one of the most plentiful polymer on the earth. Tetrahydrocurcumin (THC), a derivative of curcuma, chosen as antioxidant and antimicrobial natural substance, was added into a cellulose matrix (1.5% w/w) and the resulting material was then studied. In addition to this active compound, the paper also contained chitosan and carboxymethylcellulose, to improve the retention of THC and the mechanical properties. Mechanical, chemical and microbiological analyses were done to completely characterize the active papers. Grammage, dry and wet strength were determined. The polyphenol content was determined by the Folin-Ciolcalteu method. Antimicrobial activity of THC, in solution and after its incorporation into the papers, was tested against Gram negative bacteria (Escherichia coli and Pseudomonas putida), Gram positive bacteria (Staphylococcus aureus and Listeria innocua) and moulds (Penicillium chrysogenum and Aspergillus niger). Antioxidant capacity was also tested through the DPPH method. THC solution presented a good antioxidant capacity: it showed an EC50 equal to 4,49 ppm ( EC50 of Trolox was 2,86 ppm). Its minimal inhibitory concentration is 0,4 g/L for Gram positive bacteria, and higher than 0,6 g/L for Gram negative. THC solution was found effective in delaying the development of moulds. THC polyphenols were quantified as 0,8% on paper weight for the paper containing THC and these quantities together with chitosan resulted able to slow down the growth of microorganism. Pseudomonas is inhibited by the presence of chitosan and THC is able to amplify the antimicrobial activity of chitosan, by inhibiting also the growth of Staphylococcus and E. Coli. The presence of THC does neither affect the mechanical properties of papers, nor the color and the odor. Based on these results, it is possible to conclude that THC exhibits good antioxidant and moderate antimicrobial properties. Paper sheets didn’t lose their mechanical properties. These data will pave the way to the use of THC for the production of an active paper-based packaging to improve the shelf life of food items

Influence of substrate on beta-galactosidase production by Kluyveromyces strains

The aim of the present research was to investigate the influence of culture conditions on the levels of β-galactosidase (EC 3.2.1.23) activity produced by Kluyveromyces strains. Interest was focused on evaluating enzyme activity levels when lactose or cheese whey was employed as substrate in culture medium formulation. From an overall look at the obtained results, the tested strains were found to be able to produce β-galactosidase at promising levels. The use of cheese whey, either for strain maintenance and production trials, allowed to obtain a high cell yield associated with β-galactosidase production. The maximum β-galactosidase volumetric activity, EA max 66.5 IU/ml, corresponding to 3184 EAspec IU/g cell dw, was obtained with K. marxianus MIM 782 at 37 °C and 72 h incubation

Evaluation of the antioxidant/antimicrobial performance of Posidonia oceanica in comparison with three commercial natural extracts and as a treatment on fresh-cut peaches (Prunus persica Batsch)

This research aimed at extending the choice of natural antimicrobials/antioxidants for food applications. Four plant extracts, Posidonia oceanica (PO), Green Tea (GT), Grape seeds (GS) and Grape skin (GK), were analyzed to determine their total phenolic content, antioxidant activity and in vitro antimicrobial performance. PO extract showed the highest total phenolic content (711 mg gallic acid/g extract) and antifungal activity against Aspergillus niger and Penicillium chrysogenum. The highest antioxidant (3.81 mg/L EC50) and antibacterial activities (bactericidal against Gram positives and bacteriostatic against Gram negatives) were found for GT extract. The best performing extracts (PO and GT) were applied by dipping on peach slices in storage trials. Microbiological and pomological parameters were evaluated during 7 d storage. Total aerobic count, Pseudomonas as well as yeasts and moulds populations, were reduced by about 0.5 log cfu/g, mainly up to 5 d in all treated samples compared to the control. Total soluble solids, titratable acidity and colour (L*a*b*) changes were also delayed in treated fruit

An alternative encapsulation approach for production of active chitosan-propolis beads

Encapsulation is a promising technology to carry natural active substances, preventing their loss and maintaining their stability until use. Beads of chitosan-containing propolis have been prepared using a mono-pore filter device, which permits the encapsulation of natural polyphenols avoiding heat treatments, high shear rates and the use of toxic solvents. Beads proved to be active against Bacillis cereus, Escherichia coli, Listeria innocua, Pseudomonas fluorescens, Yarrovia lipolytica and three moulds strains; the highest effect was found against Staphylococcus aureus (MIC 0.8 mg beads mL-1). Results in liquid cultures of S. aureus evidenced that beads were able to release the flavonoids from propolis: the diffusion of the active compounds is a key factor in the exploitation of the microbial activity. The obtained chitosan-propolis beads represent an example of natural antimicrobial delivery system that could be used to prevent the growth of pathogenic/spoilage bacteria in food applications

Glutathione–enriched baker’s yeast: production, bioaccessibility and intestinal transport assays

AIMS: a glutathione (GSH) yeast-based biomass (S. cerevisiae) was used to investigate GSH stability, solubilization during gastrointestinal digestion and GSH intestinal transport. METHODS AND RESULTS: a post-growing procedure was applied to improve intracellular GSH yeast content. The presence of adenine (ADE) in the biotransformation solution (CYS-GLY-GLU mixture) and alternatively, a glucose shot after 4 h incubation, allowed to obtain cells containing about GSH 1.6-1.7% dcw (dry cell weight) (control 0.5%). Yeast samples were subjected to in vitro gastro-intestinal digestion and absorption assays employing Caco-2 and HT29-MTX cell lines in different proportions (100/0, 70/30 and 50/50). Trials were also performed to verify intestinal cell viability. CONCLUSIONS: at least 87% of ingested GSH is available in reduced form for intestinal absorption. In vitro GSH transport assays indicated that GSH is poorly absorbed (<20%). Nevertheless, studies in response to oxidative stress induced by H2 O2 demonstrated a protective role of the GSH-enriched biomass towards intestinal cell viability. SIGNIFICANCE AND IMPACT OF STUDY: an enriched GSH yeast-based biomass has been obtained using a post-growing procedure. Although GSH present in enriched yeasts is poorly absorbed by intestinal cells, this biomass showed an intestinal local protective effect, improving cells viability when a simulated oxidative stress was applied

Sakacin-A antimicrobial packaging for decreasing Listeria contamination in thincut meat: preliminary assessment

BACKGROUND: Minimally processed ready-to-eat products are considered a high-risk food because of the possibility of contamination with pathogenic bacteria, including Listeria monocytogenes from the animal reservoir, and the minimal processing they undergo. In this study, a sakacin-A anti-Listeria active package was developed and tested on thin-cut veal meat slices (carpaccio). RESULTS: Enriched food-grade sakacin-A was obtained from a cell-free supernatant of a Lactobacillus sakei culture and applied (0.63 mg cm 122) onto the surface of polyethylene-coated paper sheets to obtain an active antimicrobial package. The coating retained antimicrobial features, indicating that the process did not affect sakacin-A functionality, as evidenced in tests carried out in vitro. Thin-cut veal meat slices inoculated with Listeria innocua (a surrogate of pathogenic L. monocytogenes) were laid on active paper sheets. After 48 h incubation at 4 \ub0C, the Listeria population was found to be 1.5 log units lower with respect to controls (3.05 vs 4.46 log colony-forming units (CFU) g 121). CONCLUSION: This study demonstrates the possibility of using an antimicrobial coating containing sakacin-A to inhibit or decrease the Listeria population in ready-to-eat products, thus lowering the risk of food-related diseases

Can Zymomonas mobilis Substitute Saccharomyces cerevisiae in Cereal Dough Leavening?

Baker\u2019s yeast intolerance is rising among Western populations, where Saccharomyces cerevisiae is spread in fermented food and food components. Zymomonas mobilis is a bacterium commonly used in tropical areas to produce alcoholic beverages, and it has only rarely been considered for dough leavening probably because it only ferments glucose, fructose and sucrose, which are scarcely present in flour. However, through alcoholic fermentation, similarly to S. cerevisiae, it provides an equimolar mixture of ethanol and CO\u2082 that can rise a dough. Here, we propose Z. mobilis as a new leavening agent, as an alternative to S. cerevisiae, overcoming its technological limit with different strategies: (1) adding glucose to the dough formulation; and (2) exploiting the maltose hydrolytic activity of Lactobacillus sanfranciscensis associated with Z. mobilis. CO\u2082 production, dough volume increase, pH value, microbial counts, sugars consumption and ethanol production were monitored. Results suggest that glucose addition to the dough lets Z. mobilis efficiently leaven a dough, while glucose released by L. sanfranciscensis is not so well fermented by Z. mobilis, probably due to the strong acidification. Nevertheless, the use of Z. mobilis as a leavening agent could contribute to increasing the variety of baked goods alternative to those leavened by S. cerevisiae