Emulsifying activity of potato proteins in the presence of k-carrageenan at different pH conditions.

Oil in Water (3:1) emulsions were prepared using potato proteins in the presence or absence of 0.2% k-carrageenan at different pH conditions (3.0, 7.0, and 4.8). These emulsions showed different droplet sizes, stability, appearance, and rheological properties. The best emulsion stability was achieved combining potato proteins and k-carrageenan at pH 3.0, where uniform and small oil droplets (30 µm) were observed. The rheological properties of the emulsions were also different. The highest viscosity and G' were shown by the emulsion prepared with the addition of k-carrageenan at pH 3.0, this being attributed to the onset of a gel-like viscoelastic structure in these conditions. SDS-PAGE indicated that the superior properties of the emulsion prepared with k-carrageenan at pH 3.0 can be attributed to an electrostatic interaction between the positively charged potato proteins and the anionic polysaccharide. This interaction allowed the formation of a strong molecular network able to stabilize the system.

Increasing the Safety and Storage of Pre-Packed Fresh-Cut Fruits and Vegetables by Supercritical CO2 Process.

This work presents a feasibility lab-scale study for a new preservation method to inactivate microorganisms and increase the shelf life of pre-packed fresh-cut products. Experiments were conducted on coriander leaves and fresh-cut carrots and coconut. The technology used the combination of hydrostatic pressure (<15 MPa), low temperature (≤45 °C), and CO2 modified atmosphere packaging (MAP). The inactivation was achieved for the naturally present microorganisms (total mesophilic bacteria, yeasts and molds, total coliforms) and inoculated E. coli. Yeasts and molds and coliform were under the detection limit in all the treated samples, while mesophiles were strongly reduced, but below the detection limit only in carrots. Inoculated E. coli strains were completely inactivated (>6.0 log CFU/g) on coconut, while a reduction >4.0 log CFU/g was achieved for carrots and coriander. For all the treated products, the texture was similar to the fresh ones, while a small alteration of color was detected. Microbiological stability was achieved for up to 14 days for both fresh-cut carrots and coconut. Overall, the results are promising for the development of a new mild and innovative food preservation technique for fresh food.

Protein profile of commercial soybean milks analyzed by label-free quantitative proteomics.

The consumption of soy milk is increasing worldwide for its nutritional value and health benefits, however, its protein composition after commercialization is not well known. Technological and thermal treatments to which soy milk is subjected could affect the protein composition of the commercial products. This study compared the protein profile of 15 different commercial soy milks using a label-free quantitative proteomics approach. Proteins related to nutrient reservoir activity, endopeptidase inhibitor activity, lipid binding, and seed maturation contribute the most in terms of percentage mass. Their associated Gene Ontology terms are also enriched. Samples clustered into three groups based on their protein composition, with glycinins and beta-conglycinins being the most influential for determining the clustering. Amino acid composition estimated from the proteomics data also reflects the clustering of samples. Twenty allergenic proteins varying in abundance were identified, with Gly m 5 and Gly m 6 being the predominantly abundant allergens.

Current and future strategies for wine yeast lees valorization.

Wine lees is a sludge material mainly composed of dead yeast precipitated at the bottom of wine tanks. Along with grape pomace and grape stalks, it is one of the main by-products of the winemaking industry. Given that wine lees are considered a soil pollutant, their disposal represents a cost for wineries. Numerous wine lees recovery and valorization strategies have been proposed, with a particularly steep increase in published research in recent years. This attention is strictly linked to the concepts of circular economy and environmental sustainability that are attracting the interest of the scientific community. In this review, an overview on the available wine lees recovery and valorization strategies is reported. Additionally, the methods for the extraction and valorization of yeast's cell wall polysaccharides (ß-glucans and mannoproteins) are discussed. Finally, current and future innovative applications in different sectors of yeast ß-glucans and mannoproteins are described and critically discussed.

Research Note: Microbial inactivation of raw chicken meat by supercritical carbon dioxide treatment alone and in combination with fresh culinary herbs.

The objective of the present study was to assess the potential synergistic effect between supercritical carbon dioxide (SC-CO2) and fresh culinary herbs (Coriandrum sativum and Rosmarinus officinalis) on the microbial inactivation of raw chicken meat. The microbiological inactivation was performed on Escherichia coli and natural flora (total mesophilic bacteria, yeasts, and molds). High pressure treatments were carried out at 40°C, 80 or 140 bar from 15 to 45 min. Microbial inactivation had a strong dependence on treatment time, achieving 1.4 log CFU/g reduction of E. coli after 15 min, and up to 5 log after 45 min, while a pressure increase from 80 up to 140 bar was not significant on the microbial inactivation. Mesophilic microorganisms were strongly reduced (>2.6 log CFU/g) after 45 min, and yeasts and molds were below the detection limits of the technique (<100 CFU/g) in most cases. The combination of fresh herbs together with SC-CO2 treatment did not significantly increase the inactivation of either E. coli or natural flora, which was similar to the SC-CO2 alone. The synergistic effect was obtained on the inactivation of E. coli using a proper concentration of coriander essential oil (EO) (0.5% v/w), while rosemary EO did not show a significant effect. Color analysis after the treatment showed an increment of lightness (L*), and a decrease of redness (a*) on the surface of the sample, making the product visually similar to cooked meat. Texture analysis demonstrated the modification of the texture parameters as a function of the process pressure making the meat more similar to the cooked one.

Characterization and emulsifying properties of extracts obtained by physical and enzymatic methods from an oenological yeast strain.

BACKGROUND: Glycosylated compounds are one of the main fractions of the yeast cell wall. Thanks to their amphiphilic structure, they have been studied as stabilizers in food emulsions over a broad range of pH conditions with encouraging results. Nevertheless, extraction costs still represent an important limit for their application in the food industry. RESULTS: In this research, four extraction methods were applied to yeast cells exploiting both physical (heating and sonication) and enzymatic approaches (use of three industrial enzyme preparations, namely Glucanex®, Sur Lies and Elevage). A fifth method involving a pure ß-glucanase enzyme (Zymolyase) was taken as reference. These extraction methods were applied to the oenological strain Saccharomyces cerevisiae EC1118, and their extraction yields and chemical properties (quantitative and qualitative determination of sugars and proteins) were studied. Emulsifying activities were determined at three different pH values (3, 5 and 7). Extractions with Physical, Glucanex and Sur Lies methods were the most successful approaches to obtain relevant amounts of yeast compounds with good emulsifying activities for 2:1 oil-in-water emulsions at pH 3 and 7 over 48 h. CONCLUSIONS: These results indicate that there is the potential for the extraction approaches here proposed to become viable tools for the recovery of yeast compounds to be used as emulsifiers in foods. This approach can be considered as the starting point to explore the possibility to exploit yeast by-products from the fermentation processes (e.g. fermentation lees from wine and beer making) as valuable compounds for food applications. © 2019 Society of Chemical Industry.

A Novel Method for the Quantification of White Wine Mannoproteins by a Competitive Indirect Enzyme-Linked Lectin Sorbent Assay (CI-ELLSA).

Mannoproteins (MPs) are cell wall proteoglycans released in wine by yeast during fermentation and ageing on lees, a procedure used for the production of several wines to enrich them in these components with consequences from both a technological and sensory point of view. Given the significance that wine MPs have for wine quality, winemakers would welcome a simple and accurate method for their quantification, as this would allow them to have a better control of this aspect at different winemaking stages. This study develops and validates a novel, simple and accurate method for MPs quantification in white wines based on a competitive indirect enzyme-linked lectin sorbent assay (CI-ELLSA), using the highly mannosylated yeast invertase as the standard. The method utilizes the lectin concanavalin A (ConA) as the immobilized ligand for MPs, and peroxidase, an enzyme rich in mannose, as the competitor for ConA. After addition of the peroxidase substrate, the intensity of the signal produced by the activity of this enzyme (absorbance at 450 nm) is inversely proportional to the amount of mannosylated proteins in the sample. Results have been validated on several wine styles including still, sparkling and sweet wines.

Nuclease expression in efficient polyhydroxyalkanoates-producing bacteria could yield cost reduction during downstream processing.

Industrial manufacturing of polyhydroxyalkanoates (PHAs) requires purification of PHAs granules from high-cell-density cultures. Cells are broken by homogenization and PHAs granules are cleansed and treated to obtain PHAs latexes. However, cell lysis releases large amounts of DNA which results in an increasing viscosity of the medium, hampering the following downstream steps. Drop in viscosity is generally achieved by costly procedures such as heat treatment or the supplementation of hypochlorite and commercially available nucleases. Searching for a cost-effective solution to this issue, a nuclease gene from Staphylococcus aureus has been integrated into two efficient PHAs-producing bacteria: Cupriavidus necator DSM 545 and Delftia acidovorans DSM 39. Staphylococcal nuclease has been proficiently expressed in both microbial hosts without affecting PHAs production. Moreover, the viscosity of the lysates of recombinant C. necator cells was greatly reduced, indicating that the engineered strain is expected to yield large reduction cost in PHAs downstream processing.

Garlic (Allium sativum L.) fed to dairy cows does not modify the cheese-making properties of milk but affects the color, texture, and flavor of ripened cheese.

Garlic and garlic components have recently been proposed as ruminal activity modulators to reduce the enteric methane emissions of ruminants, but little is known of their influence on milk coagulation properties, nutrient recovery, cheese yield, and sensorial and rheological characteristics of milk and cheese. The present study assessed the effects of garlic and diallyl sulfide supplements on dry matter intake (DMI), productive performance, milk coagulation properties, cheese yield, milk and cheese sensory profiles, and rheological characteristics. Four dairy cows were fed a total mixed ration either alone (control) or supplemented with 100 or 400 g/d of garlic cloves or 2 g/d of diallyl sulfide in 4 consecutive experimental periods in a 4 × 4 Latin square design. The diallyl sulfide dose was established to provide approximately the same amount of allyl thiosulfinate compounds as 100 g of fresh garlic cloves. The total mixed ration was composed of 0.29 corn silage, 0.23 corn-barley mixture, 0.17 sunflower-soybean mixture, 0.12 alfalfa hay, 0.12 grass hay, 0.04 sugar beet pulp, and 0.02 other additives, and contained 0.253 starch, 0.130 crude protein, and 0.375 neutral detergent fiber, on a dry matter basis. Each experimental period consisted of 7 d of transition and 14 d of treatment. On d 18 and 21 of each period, milk samples (10 L) were collected from each cow for chemical analysis and cheese-making. The organoleptic properties of the milk and 63-d-ripened cheeses were assessed by a panel of 7 trained sensory evaluators. The experimental treatments had no effects on DMI, milk yield, feed efficiency (milk yield/DMI), milk coagulation properties, nutrient recovery, or cheese yield. Garlic-like aroma, taste, and flavor of milk and cheese were significantly influenced by the treatments, particularly the highest dose of garlic cloves, and we found close exponential relationships between milk and cheese for garlic-like aroma (R2 = 0.87) and garlic-like flavor (R2 = 0.79). Diallyl sulfide and 400 g/d of garlic cloves resulted in lower pH, shear force, and shear work of ripened cheeses compared with the other treatments. Garlic cloves and diallyl sulfide had opposite effects on cheese color indices. We conclude that adding 400 g/d of garlic to the feed of lactating dairy cows highly influences the sensory and rheological characteristics of cheese.

Exploring the use of Saccharomyces cerevisiae commercial strain and Saccharomycodes ludwigii natural isolate for grape marc fermentation to improve sensory properties of spirits.

In Mediterranean countries the most diffuse practice to obtain the valorization of grape marc, the main by-product from winemaking, is the production of spirits. During this process, marc storage for sugar fermentation represents a crucial step, since side-fermentations leading to off-flavours production can very easily occur. In this study we evaluated the effect of the addition of two yeast strains, inoculated separately at the beginning of the storage period, into marcs from two Italian grape varieties with the aim to control the development of autochthonous microbiota and to improve spirit quality. The presence of the inoculated strains was monitored by means of PCR-based approaches. A commercial Saccharomyces cerevisiae strain, chosen as this species is notably the best ethanol producer, showed excellent ability to dominate the autochthonous microflora and to reduce off-flavours as demonstrated by chemical analysis and sensory evaluation. A Saccharomycodes ludwigii strain, chosen for increasing varietal compounds thus enhancing spirit aroma, showed a level of implantation not sufficient to assure a clear beneficial effect on quality. The implantation level of this strain was affected by S. cerevisiae competition since the highest level was found in grape marc with lower sugar content, where indigenous S. cerevisiae were less persistent.

Antioxidant activity of bovine casein hydrolysates produced by Ficus carica L.-derived proteinase.

A Ficus carica L. latex proteinase preparation was investigated for its ability to produce antioxidant hydrolysates/peptides from bovine casein (CN). The Oxygen Radical Absorbance Capacity (ORAC) values for NaCN and ß-CN hydrolysates ranged from 0.06 to 0.18, and from 0.51 to 1.19µmol Trolox equivalents/mg freeze-dried sample, respectively. Gel permeation HPLC showed that the ß-CN hydrolysate with a degree of hydrolysis of 21% had 65% of peptide material with a molecular mass <500Da. The RP-UPLC profiles also indicated that ß-CN was substantially hydrolysed during the early stages of hydrolysis. Analysis of the 4h ß-CN hydrolysate by LC-ESI-MS/MS allowed identification of 8 peptide sequences with potential antioxidant properties.

Antibrowning potential of Brassicacaea processing water.

The effects of Brassicacaea processing water on polyphenol oxidases from different plant sources were investigated. Results showed that processing water (PW) prepared by cooking young Brassicacaea leaves with water has the capacity to inhibit both commercial tyrosinase and plant polyphenol oxidases. PW was freeze-dried (LPW) in an attempt to increase its utility and improve its efficiency at inhibition of polyphenol oxidases, however this resulted in a loss of activity, probably due to the stresses of the freezing and/or drying processes. The addition of PW to ascorbic acid resulted in complete inhibition of grape polyphenol oxidase, suggesting that the combination of the two antibrowning agents may be a promising tool to control this enzymatic activity.

3,4-Dihydroxyphenylalanine gel diffusion assay for polyphenol oxidase quantification.

We have developed a simple, inexpensive plate assay to detect and quantify polyphenol oxidase (PPO) activity from different origins. The logarithm of enzyme activity is linearly correlated with the diameter of the dark, l-3,4-dihydroxyphenylalanine (l-DOPA) oxidized circles produced in the gel, thereby allowing quantification of PPO. Moreover, precision and high reproducibility of the assay were confirmed by statistical analysis.

Deriphat 2-DE to visualize polyphenol oxidase in Moscato and Prosecco grapes.

The Deriphat 2-DE was used to visualize polyphenol oxidase (PPO) isoforms of Moscato and Prosecco grape extracts, partially purified and characterized. Catecholase has similar values in the two varieties, whereas Moscato cresolase data are almost 54% higher. In the first dimension, the PPO of both varieties may be detected by SDS-PAGE, but native PAGE (N-PAGE) gave negative results. For this reason, the samples were solubilized in the zwitteronic detergent Deriphat, which was also included in the gel and the cathodic buffer. Deriphat migrated together with the cathodic buffer, maintaining protein solubility and revealing the PPO profiles of Moscato and Prosecco extracts in native conditions. The combination of Deriphat-PAGE (D-PAGE) and SDS-PAGE (2-DE) also resulted in improved separation efficiency in resolving PPO and specialized stains in evaluating PPO activities. The control, represented by IEF for the first-dimensional separation, had a lower number of spots, demonstrating the higher capacity of Deriphat 2-DE to isolate PPO isoforms from grape extracts. The Deriphat 2-DE method described here is simple but powerful, and the resulting information will be a useful tool for further proteomic research.

Protein haze formation in white wines: effect of saccharomyces cerevisiae cell wall components prepared with different procedures.

Cell wall material was extracted by five different methods from an oenological strain of Saccharomyces cerevisiae. Enzyme preparations containing beta-glucanase activity (Zymolyase, Glucanex, and Finizym 250L) allowed a better extraction yield compared to that of dithiothreitol (DTT) and EDTA. The yeast extracts were only soluble in part in wine. The wine-soluble fraction (WSF) of the five extracts, differing in both protein and sugar contents, when added in increasing amounts to white wine differently affected protein haze formation, as determined by the heat test, giving dose/response curves of different shapes. The curves obtained with the WSF derived from DDT and Zymolyase extracts showed a plateau value corresponding to 90% and 80% of wine haze reduction, respectively. In contrast, addition of the WSF derived from the other extracts resulted in increased turbidity with respect to the original wine. The mannoproteins (MP), isolated from each yeast extract by Concanavalin-A chromatography, gave dose/response curves showing shapes more similar among them than those obtained with the whole WSFs. The best wine stabilization was obtained with the MP of the DTT and Zymolyase extracts. The supernatants obtained after heating the MP of the different extracts were also tested. The stabilizing effect of the heat-stable MP (HSMP) was always larger than that of the corresponding total (un-heated) MP. The HSMP obtained starting from the DTT and Zymolyase extracts showed the best haze-protecting effect, which was, however, lower than that obtained with their corresponding WSF. This result suggests that wine protein stabilization by compounds of the yeast cell wall could be related, in addition to the action of the MP, also to the presence of other substances of different nature.