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11 pages, 2224 KiB  
Article
Color Stability of Various Orthodontic Clear Aligner Systems after Submersion in Different Staining Beverages
by Nicolae Daniel Olteanu, Ionut Taraboanta, Tinela Panaite, Carina Balcos, Sorana Nicoleta Rosu, Raluca Maria Vieriu, Stefania Dinu and Irina Nicoleta Zetu
Materials 2024, 17(16), 4009; https://doi.org/10.3390/ma17164009 - 12 Aug 2024
Abstract
This study aimed to compare the color changes in two different orthodontic clear aligner systems after submersion in various beverages for 14 days. The tested aligner systems were Taglus Premium made of polyethylene terephthalate glycol (the TAG group) and CA® Prodin+ made [...] Read more.
This study aimed to compare the color changes in two different orthodontic clear aligner systems after submersion in various beverages for 14 days. The tested aligner systems were Taglus Premium made of polyethylene terephthalate glycol (the TAG group) and CA® Prodin+ made of a transparent copolyester and a thermoplastic elastomer (the PRO group). A total of 56 samples were firstly divided into two groups according to the tested system—TAG and PRO. Each group was subsequently divided in four subgroups according to immersion solution: A—artificial saliva, B—cola, C—coffee, D—red wine. Color measurements were performed on Days 1, 7 and 14 using a portable colorimeter and the CIE L*a*b* system. The obtained results showed significant color changes in both materials when exposed to coffee and red wine (p > 0.05). Samples in the PRO group showed a greater susceptibility to discoloration (higher ΔE values) when compared to the TAG group after submersion in cola (p = 0.025), coffee (p = 0.005) and red wine (p = 0.041) solutions. Statistical analysis revealed that all of the color parameters ΔL*, Δa*, Δb* and ΔE of both tested materials were affected by submersion in coffee solution for 14 days. In conclusion, the CA® Pro+ aligner system is more prone to staining compared to the Taglus material after submersion for 14 days in cola, coffee and red wine solutions. Submersion for 14 days in coffee solution alters all of the color parameters (ΔL, Δa, Δb and ΔE) of both tested aligner materials. Full article
(This article belongs to the Special Issue Advanced Dental Materials: From Design to Application)
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<p>Study design and distribution of samples in groups and subgroups.</p>
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<p>Mean values and standard deviations of the ΔL* parameter within each study stage for the TAG and PRO groups.</p>
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<p>Mean values and standard deviations of the Δa* parameter within each study stage for the TAG and PRO groups.</p>
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<p>Mean values and standard deviations of the Δb* parameter within each study stage for the TAG and PRO groups.</p>
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<p>Box-plot representation of the distribution of ΔE values of each group and subgroup on each test day.</p>
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9 pages, 724 KiB  
Brief Report
Cell Wall Profiling of the Resurrection Plants Craterostigma plantagineum and Lindernia brevidens and Their Desiccation-Sensitive Relative, Lindernia subracemosa
by John P. Moore, Brock Kuhlman, Jeanett Hansen, Leonardo Gomez, Bodil JØrgensen and Dorothea Bartels
Plants 2024, 13(16), 2235; https://doi.org/10.3390/plants13162235 - 12 Aug 2024
Abstract
Vegetative desiccation tolerance has evolved within the genera Craterostigma and Lindernia. A centre of endemism and diversification for these plants appears to occur in ancient tropical montane rainforests of east Africa in Kenya and Tanzania. Lindernia subracemosa, a desiccation-sensitive relative of Craterostigma [...] Read more.
Vegetative desiccation tolerance has evolved within the genera Craterostigma and Lindernia. A centre of endemism and diversification for these plants appears to occur in ancient tropical montane rainforests of east Africa in Kenya and Tanzania. Lindernia subracemosa, a desiccation-sensitive relative of Craterostigma plantagineum, occurs in these rainforests and experiences adequate rainfall and thus does not require desiccation tolerance. However, sharing this inselberg habitat, another species, Lindernia brevidens, does retain vegetative desiccation tolerance and is also related to the resurrection plant C. plantagineum found in South Africa. Leaf material was collected from all three species at different stages of hydration: fully hydrated (ca. 90% relative water content), half-dry (ca. 45% relative water content) and fully desiccated (ca. 5% relative water content). Cell wall monosaccharide datasets were collected from all three species. Comprehensive microarray polymer profiling (CoMPP) was performed using ca. 27 plant cell-wall-specific antibodies and carbohydrate-binding module probes. Some differences in pectin, xyloglucan and extension epitopes were observed between the selected species. Overall, cell wall compositions were similar, suggesting that wall modifications in response to vegetative desiccation involve subtle cell wall remodelling that is not reflected by the compositional analysis and that the plants and their walls are constitutively protected against desiccation. Full article
(This article belongs to the Special Issue New Perspectives on the Plant Cell Wall)
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Graphical abstract

Graphical abstract
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<p>Monosaccharide compositional analysis of the total AIR isolated from leaf material of <span class="html-italic">Craterostigma plantagineum</span> (<b>a</b>), and <span class="html-italic">Lindernia brevidens</span> (<b>b</b>). White bars represent hydrated leaves, mid-grey shaded bars represent partially hydrated leaves and shaded bars represent desiccated leaves. Monosaccharide codes are for arabinose (Ara), rhamnose (Rha), fucose (Fuc), xylose (Xyl), mannose (Man), galactose (Gal), galacturonic acid (GalUA), glucose (Glc) and glucuronic acid (GlcUA). Error bars represent the standard error (SE) of the mean of four biological samples with two technical replicates per biological sample. Statistically significant differences, based on one-way ANOVA variance testing, are indicated on the bar graphs as an asterisk.</p>
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<p>Comprehensive microarray polymer profiling (CoMPP) analysis of plant-leaf cell wall fractions from CDTA-extractable material (<b>a</b>) and NaOH-extractable material (<b>b</b>) isolated from <span class="html-italic">Craterostigma plantagineum</span>, <span class="html-italic">Lindernia brevidens</span> and <span class="html-italic">Lindernia subracemosa</span> leaves that were hydrated (H), partially hydrated (PD) or desiccated (D). The heatmaps indicate the relative abundance of plant cell wall glycan-associated epitopes present in the AIR, and the colour intensity is correlated to the mean spot signals. The values in the heatmap are the mean spot signals from three experiments. The highest signal in the entire data set was set to 100, and all other data were adjusted accordingly.</p>
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21 pages, 3062 KiB  
Article
Bioprocess Optimization by Taguchi Design and Response Surface Analysis for Obtaining Active Yeast Used in Vinification
by Corina Dumitrache, Mihaela Violeta Ghica, Mihai Frîncu, Iuliana Diana Bărbulescu, Mihaela Begea, Camelia Filofteia Diguță, Cornel Baniță, Valeriu V. Cotea, Florentina Israel-Roming and Răzvan Ionuț Teodorescu
Fermentation 2024, 10(8), 413; https://doi.org/10.3390/fermentation10080413 - 9 Aug 2024
Viewed by 226
Abstract
This study presents the behavior of the identified yeast strain S. cerevisiae, isolated from Busuioacă de Bohotin grapes from the Pietroasa winery, during the process of fermentation to obtain dry active yeast biomass for the winemaking process. In this respect, we promoted [...] Read more.
This study presents the behavior of the identified yeast strain S. cerevisiae, isolated from Busuioacă de Bohotin grapes from the Pietroasa winery, during the process of fermentation to obtain dry active yeast biomass for the winemaking process. In this respect, we promoted an optimization strategy for obtaining active dry yeast biomass. The cultivation conditions for micropilot fermentation (temperature, pH, carbon source, and nitrogen source) were selected and designed according to a Taguchi design with four factors and three levels. Reproducibility testing was conducted under specific fermentation parameters: temperature (32 °C), pH (4.5), carbon source (12%), and nitrogen source (0.7%). Following the optimization process, two combinations of cultivation parameters were selected, and one of them, based on the results, was selected for further analysis. Quantitative data were obtained, showing dry yeast biomass (DCW) at 1.39 g/100 mL and protein content at 45.57%. The active yeast was then used in the winemaking process for Tămâioasă Românească and Busuioacă de Bohotin varieties at Pietroasa winery for validation. This optimization aims to facilitate easy and rapid production of fresh wine yeast tailored to the local winemaking practices of Pietroasa winery, with real application potential in other viticultural areas, aligning with the terroir concept. Full article
(This article belongs to the Section Microbial Metabolism, Physiology & Genetics)
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<p>Yeast biomass after freeze-drying process.</p>
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<p>Biotechnological process for yeast biomass.</p>
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<p>(<b>a</b>,<b>b</b>). Plots showing the correlation between experimental and theoretical values for (<b>a</b>) dry biomass (g/100 mL) and (<b>b</b>) protein content (%).</p>
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<p>(<b>a</b>,<b>b</b>). Plots showing the correlation between expected normal values and residuals for (<b>a</b>) dry biomass (g/100 mL) and (<b>b</b>) protein content (%).</p>
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<p>Three-dimensional response surface and contour levels indicating the effect of different cultivation parameters on dry biomass response (Y<sub>1</sub>): (<b>a</b>) temperature (X<sub>1</sub>) and pH (X<sub>2</sub>); (<b>b</b>) temperature (X<sub>1</sub>) and carbon source (X<sub>3</sub>); (<b>c</b>) temperature (X<sub>1</sub>) and yeast extract (X<sub>4</sub>); (<b>d</b>) pH (X<sub>2</sub>) and carbon source (X<sub>3</sub>); (<b>e</b>) pH (X<sub>2</sub>) and yeast extract (X<sub>4</sub>); (<b>f</b>) carbon source (X<sub>3</sub>) and yeast extract (X<sub>4</sub>). The dark red color refers to the maximum values for the dependent variable Y1, and the green color to their minimum values.</p>
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<p>Three-dimensional response surface and contour levels indicating the effect of different cultivation parameters on the protein content response (Y<sub>2</sub>): (<b>a</b>) temperature (X<sub>1</sub>) and pH (X<sub>2</sub>); (<b>b</b>) temperature (X<sub>1</sub>) and carbon source (X<sub>3</sub>); (<b>c</b>) temperature (X<sub>1</sub>) and yeast extract (X<sub>4</sub>); (<b>d</b>) pH (X<sub>2</sub>) and carbon source (X<sub>3</sub>); (<b>e</b>) pH (X<sub>2</sub>) and yeast extract (X<sub>4</sub>); (<b>f</b>) carbon source (X<sub>3</sub>) and yeast extract (X<sub>4</sub>). The dark red color refers to the maximum values for the dependent variable Y2, and the green color to their minimum values.</p>
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<p>Effects of controlled factors on S/N ratio for the dependent variable: (<b>a</b>) Y<sub>1</sub>—dry biomass (%); (<b>b</b>) Y<sub>2</sub>—protein content.</p>
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<p>Sensorial evaluation of Busuioacă de Bohotin wines.</p>
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<p>Sensorial evaluation of Tămâioasă Românească wines.</p>
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18 pages, 5372 KiB  
Article
Elucidating the Subcellular Localization of GLRaV-3 Proteins Encoded by the Unique Gene Block in N. benthamiana Suggests Implications on Plant Host Suppression
by Patrick Lameront, Mehdi Shabanian, Laura M. J. Currie, Catherine Fust, Caihong Li, Alyssa Clews and Baozhong Meng
Biomolecules 2024, 14(8), 977; https://doi.org/10.3390/biom14080977 - 9 Aug 2024
Viewed by 410
Abstract
Grapevine leafroll-associated virus 3 (GLRaV-3) is a formidable threat to the stability of the global grape and wine industries. It is the primary etiological agent of grapevine leafroll disease (GLD) and significantly impairs vine health, fruit quality, and yield. GLRaV-3 is a member [...] Read more.
Grapevine leafroll-associated virus 3 (GLRaV-3) is a formidable threat to the stability of the global grape and wine industries. It is the primary etiological agent of grapevine leafroll disease (GLD) and significantly impairs vine health, fruit quality, and yield. GLRaV-3 is a member of the genus Ampelovirus, Closteroviridae family. Viral genes within the 3′ proximal unique gene blocks (UGB) remain highly variable and poorly understood. The UGBs of Closteroviridae viruses include diverse open reading frames (ORFs) that have been shown to contribute to viral functions such as the suppression of the host RNA silencing defense response and systemic viral spread. This study investigates the role of GLRaV-3 ORF8, ORF9, and ORF10, which encode the proteins p21, p20A, and p20B, respectively. These genes represent largely unexplored facets of the GLRaV-3 genome. Here, we visualize the subcellular localization of wildtype and mutagenized GLRaV-3 ORFs 8, 9, and 10, transiently expressed in Nicotiana benthamiana. Our results indicate that p21 localizes to the cytosol, p20A associates with microtubules, and p20B is trafficked into the nucleus to carry out the suppression of host RNA silencing. The findings presented herein provide a foundation for future research aimed at the characterization of the functions of these ORFs. In the long run, it would also facilitate the development of innovative strategies to understand GLRaV-3, mitigate its spread, and impacts on grapevines and the global wine industry. Full article
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<p>Genome structure of grapevine leafroll associated virus 3 (GLRaV-3) and protein expression constructs. (<b>A</b>). Genome structure of GLRaV-3. The RNA genome contains 12–13 open reading frames (ORFs). ORF1a/b encodes a replicase polyprotein. ORFs 3–7 make up the “Quintuple Gene Block” (QGB) shared among the family <span class="html-italic">Closteroviridae</span>, which encode five polypeptides designated p5, Hsp70h, p55, the coat protein, and the coat protein minor. ORFs 8–12 make up the “Unique Gene Block” (UGB), which encodes the polypeptides p21, p20A, p20B, p4, and p7. (<b>B</b>). Graphic representation of the viral protein expression constructs. All constructs were made based on pRTL3-GFP and pRTL3-mRFP. Each contains the cauliflower mosaic virus (CaMV) 35S promoter, the tobacco etch virus translation enhancer (TE), and the CaMV 35S termination signal (TERM). Restriction sites used to make these constructs are given above each construct. GFP: green fluorescent protein; mRFP: monomeric red fluorescent protein. (<b>C</b>). Graphic representation of the organelle and cytoskeletal fluorescent markers used in this study.</p>
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<p>In silico analysis of the predicted structure and subcellular localization of p21 using CLSM. In silico protein structural prediction and analysis of p21 as predicted by ColabFold. (<b>A</b>). Surface electrostatics of p21 generated by APBS (Jurrus E et al.). (<b>B</b>). Cartoon representation of predicted p21 structure. N and C termini are labeled. (<b>C</b>). <span class="html-italic">N. benthamiana</span> agro-infiltrated with p21-EGFP. (<b>D</b>). <span class="html-italic">N. benthamiana</span> agro-infiltrated with p21-EGFP and mRFP.</p>
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<p>In silico analysis of the predicted structure and subcellular localization of p20A using CLSM. (<b>A</b>). Predicted structure of p20A, colored by electrostatics generated by APBS. (<b>B</b>). Cartoon representation of the predicted structure of p20A. Positively charged residues are colored blue. N and C termini are labeled. (<b>C</b>). <span class="html-italic">N. benthamiana</span> leaf tissue agro-infiltrated with CD3-987 and p20A-mRFP. Green and red channels are overlayed in the rightmost panel. (<b>D</b>). <span class="html-italic">N. benthamiana</span> leaf tissue agro-infiltrated with mTalin-GFP and p20A-mRFP. Green and red channels are overlayed in the rightmost panel. (<b>E</b>). <span class="html-italic">N. benthamiana</span> leaf tissue agro-infiltrated with MBD-GFP and p20A-mRFP. Green and red channels are overlayed in the rightmost panel. White triangles indicate examples of colour shift in overlayed image.</p>
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<p>In silico analysis of the predicted structure and subcellular localization of p20B using CLSM. (<b>A</b>). Predicted structure of p20B, colored by electrostatics calculated by APBS. (<b>B</b>). Cartoon representation of the predicted structure of p20B. Predicted nuclear localization signal is colored in blue and highlighted. N and C termini are labeled. (<b>C</b>). Chloroplast autofluorescence captured from <span class="html-italic">N. benthamiana</span>. agro-infiltrated with p20B-EGFP. Chloroplast autofluorescence and green channels are overlayed in the rightmost panel. (<b>D</b>). <span class="html-italic">N.benthamiana</span> leaf tissue infiltrated with pCP:mRFP and p20B-EGFP. Chloroplast autofluorescence; red and green channels are overlayed in the rightmost panel.</p>
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<p>p20B GUS tagging, p20B NLS mutagenesis, and p20B NLS tagging of subcellular localization analysis using CLSM. (<b>A</b>). <span class="html-italic">N. benthamiana</span> agro-infiltrated with pCP:mRFP and p20B-EGFP-GUS. Chloroplast autofluorescence; red and green channels are overlayed in the rightmost panel. (<b>B</b>). <span class="html-italic">N. benthamiana</span> agro-infiltrated with pCP:mRFP and p20B-ΔNLS2-EGFP-GUS. Chloroplast autofluorescence, red and green channels are overlayed in the rightmost panel. (<b>C</b>). <span class="html-italic">N. benthamiana</span> agro-infiltrated with pCP:mRFPand p20B-ΔNLSε-EGFP-GUS. Chloroplast autofluorescence; red and green channels are overlayed in the rightmost panel. (<b>D</b>). <span class="html-italic">N. benthamiana</span> agro-infiltrated with pCP:mRFP and NLS-EGFP-GUS. Chloroplast autofluorescence, red and green channels are overlayed in the rightmost panel.</p>
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16 pages, 1037 KiB  
Article
Foliar Co-Applications of Nitrogen and Iron on Vines at Different Developmental Stages Impacts Wine Grape (Vitis vinifera L.) Composition
by Xiaoke Fu, Xi Chen, Yiwen Chen, Yueran Hui, Rui Wang and Yaqi Wang
Plants 2024, 13(16), 2203; https://doi.org/10.3390/plants13162203 - 9 Aug 2024
Viewed by 214
Abstract
The co-application of N and Fe can improve wine grape composition and promote the formation of flavor compounds. To understand the effects of foliar co-application of N and Fe on wine grape quality and flavonoid content, urea and EDTA-FE were sprayed at three [...] Read more.
The co-application of N and Fe can improve wine grape composition and promote the formation of flavor compounds. To understand the effects of foliar co-application of N and Fe on wine grape quality and flavonoid content, urea and EDTA-FE were sprayed at three different developmental stages. Urea and EDTA-Fe were sprayed during the early stage of the expansion period, at the end of the early stage of the expansion period to the late stage of the veraison period, and during the late stage of the veraison period. The results demonstrated that the co-application of urea and EDTA-Fe, particularly N application during the late stage of the veraison period and Fe application during the early stage of the berry expansion period (N3Fe1), significantly improved grape quality. Specifically, the soluble solid content of berries increased by 2.78–19.13%, titratable acidity decreased by 6.67–18.84%, the sugar-acid ratio became more balanced, and yield increased by 13.08–40.71%. Further, there was a significant increase in the relative content of amino acids and flavonoids. In conclusion, the application of Fe and N fertilizers at the pre-expansion and late veraison stages of grapes can significantly improve the quality and yield of berries; ultimately, this establishes a foundation for future improvement in the nutritional value of grapes and wine. Full article
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<p>Chlorophyll content in wine grape leaves treated with co-applications of N and Fe. Effect of N and Fe co-application on the chlorophyll of wine grape leaves. Different lowercase letters (abcd) indicate significant differences (<span class="html-italic">p</span> &lt; 0.05).</p>
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<p>Effect of co-application of N, Fe, and leaves/petiole on N and Fe contents in leaves and petioles of wine grapes at different developmental stages. Different lowercase letters indicate significant differences (<span class="html-italic">p</span> &lt; 0.05).</p>
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16 pages, 1297 KiB  
Article
Polyphenols Extraction from Different Grape Pomaces Using Natural Deep Eutectic Solvents
by Alessandro Frontini, Andrea Luvisi, Carmine Negro, Massimiliano Apollonio, Rita Accogli, Mariarosaria De Pascali and Luigi De Bellis
Separations 2024, 11(8), 241; https://doi.org/10.3390/separations11080241 - 8 Aug 2024
Viewed by 281
Abstract
Exploiting by-products from the oenological industry to extract antioxidant chemicals is a shared goal that combines the need to reduce the wine sector’s environmental impact with the need to improve the availability of these biomolecules, according to a circular economy approach. Natural deep [...] Read more.
Exploiting by-products from the oenological industry to extract antioxidant chemicals is a shared goal that combines the need to reduce the wine sector’s environmental impact with the need to improve the availability of these biomolecules, according to a circular economy approach. Natural deep eutectic solvents (NaDES) have recently captured researchers’ interest as a safer and more environmentally friendly alternative to traditional solvents due to their effectiveness, low toxicity, and stability. In this work, we set out to investigate several NaDES for the extraction of phenolic chemicals from local monovarietal grape pomace resulting from different vinification procedures (including both red and rosé vinification of Negroamaro and Primitivo grapes; rosé vinification of Susumaniello grapes and white vinification of Chardonnay, Fiano and Malvasia bianca grapes), with the additional goal of generalizing the use of NaDES to extract chemicals of interest from organisms selected from the wide plant biodiversity. Three binary choline chloride-based NaDES (DES-Lac, DES-Tar, and DES-Gly, with lactic acid, tartaric acid, and glycerol as hydrogen bond donors, respectively) were compared to ethanol as a conventional solvent, and the extracts were evaluated using HPLC/MS and colorimetric techniques. The results revealed that each NaDES produces a substantially higher total phenolic yield than ethanol (up to 127.8 mg/g DW from Primitivo rosé grape pomace). DES-Lac and DES-Tar were more effective for anthocyanins extraction; the most abundant compound was malvidin 3-O-glucoside (highest extraction yield with DES-Lac from Susumaniello pomace: 29.4 mg/g DW). Regarding phenolic compounds, DES-Gly was the most effective NaDES producing results comparable to ethanol. Unexpectedly, Chardonnay pomace has the greatest content of astilbin. In most cases, grape pomace extracts obtained by rosé and white vinification provided the maximum yield. As a result, NaDES have emerged as a viable alternative to traditional organic solvent extraction techniques, allowing for higher (or equal) yields while significantly lowering costs, hazards, and environmental impact. Full article
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<p>Total phenolic content, expressed as mg of gallic acid equivalent (GAE) per gram (dry weight) of pomace. For each sample, a one-way ANOVA test was performed to assess the statistically significant difference among different solvents tested, followed by Tukey post hoc test (HSD); for each pomace sample, bars with different letters differ at <span class="html-italic">p</span> &lt; 0.05.</p>
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<p>Representative HPLC/DAD chromatograms at λ = 520 nm. (<b>a</b>) Anthocyanins extracted with DES-Lac from Susumaniello rosé grape pomace; (<b>b</b>) anthocyanins extracted with ethanol from Susumaniello rosé grape pomace. The capital letters above each peak indicate the chemical compound as shown in <a href="#separations-11-00241-t003" class="html-table">Table 3</a>.</p>
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<p>HPLC/DAD chromatograms at λ = 280 nm. (<b>a</b>) Phenolic compounds extracted with DES-Gly from Primitivo rosé grape pomace; (<b>b</b>) Phenolic compounds extracted with ethanol from Primitivo rosé grape pomace. The numbers above each peak indicate the chemical compound as shown in <a href="#separations-11-00241-t005" class="html-table">Table 5</a>.</p>
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12 pages, 825 KiB  
Article
Production of Functional Vinegar Enriched with γ-Aminobutyric Acid through Serial Co-Fermentation of Lactic Acid and Acetic Acid Bacteria Using Rice Wine Lees
by Yun-Ho Park, Min-Jeong Kwon, Dong-Min Shin and Sam-Pin Lee
Appl. Microbiol. 2024, 4(3), 1203-1214; https://doi.org/10.3390/applmicrobiol4030082 - 8 Aug 2024
Viewed by 219
Abstract
Functional vinegar with high γ-aminobutyric acid (GABA) content was manufactured through a two-stage serial co-fermentation of rice wine lees, a by-product of Korean rice wine, using lactic acid bacteria (LAB) and acetic acid bacteria (AAB). The first LAB fermentation elevated GABA content by [...] Read more.
Functional vinegar with high γ-aminobutyric acid (GABA) content was manufactured through a two-stage serial co-fermentation of rice wine lees, a by-product of Korean rice wine, using lactic acid bacteria (LAB) and acetic acid bacteria (AAB). The first LAB fermentation elevated GABA content by utilizing monosodium glutamate (MSG) as a precursor. Lactiplantibacillus plantarum KS2020 converted up to 10% of MSG into GABA and indicated a GABA content of 65.49 mg/g. The concentration of LAB-fermented rice wine lees was then optimized for the second co-fermentation, and Acetobacter aceti was used to produce vinegar. Co-fermentation using 40% first LAB-fermented rice wine lees yielded vinegar with 55.34 mg/g acetic acid and 22.61 mg/g GABA. The temperature-dependent reduction in GABA in GABA-enriched vinegar followed the Arrhenius relationship during storage, with an activation energy of 9.94 kcal/mol (20–35 °C, R2 = 0.99). The GABA present in the vinegar showed evidence of a temperature-/time-dependent decrease, decreasing by 40% over five months. This study first proved the higher GABA-enriched vinegar production from rice wine lees using Lb. plantarum KS2020 and A. aceti. Full article
(This article belongs to the Special Issue Applied Microbiology of Foods 2.0)
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<p>Viable bacterial count, pH, acidity, and TLC of rice wine lees fermented by <span class="html-italic">Lb. plantarum</span> KS 2020. (<b>A</b>) Viable bacterial count of <span class="html-italic">Lb. plantarum</span> in first LAB fermentation. (<b>B</b>) pH of first LAB fermentation. (<b>C</b>) Acidity of first LAB fermentation. (<b>D</b>) TLC of first LAB fermentation. <sup>(1)</sup> A: 8% MSG; <sup>(2)</sup> B: 10% MSG; <sup>(3)</sup> C: 12% MSG. MSG: Monosodium L-glutamate. The error bars indicate standard deviations. Different letters denote statistical difference (<span class="html-italic">p</span> &lt; 0.05). n = 3 for each group.</p>
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<p>Viable bacterial count, pH, and acidity of <span class="html-italic">Lb. plantarum</span> and <span class="html-italic">A. aceti</span> in co-fermented rice wine lees vinegar by <span class="html-italic">Lb. plantarum</span> KS 2020 and <span class="html-italic">A. aceti</span>. (<b>A</b>) Viable bacterial count of <span class="html-italic">Lb. plantarum</span> in second fermentation. (<b>B</b>) Viable bacterial count of <span class="html-italic">A. aceti</span> in second fermentation. (<b>C</b>) Acidity in second fermentation. <sup>(1)</sup> B1: First fermented rice wine lees (40%); <sup>(2)</sup> B2: first fermented rice wine lees (45%); <sup>(3)</sup> B3: first fermented rice wine lees (50%). LAB fermentation was performed with rice wine lees containing 10% MSG. The error bars indicate standard deviations. Different letters denote statistical difference (<span class="html-italic">p</span> &lt; 0.05). n = 3 for each group.</p>
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<p>GABA stability test modeling in rice wine lees vinegar co-fermented with <span class="html-italic">Lb. plantarum</span> KS2020 and <span class="html-italic">A. aceti</span> using the Arrhenius equation.</p>
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13 pages, 5120 KiB  
Article
The Metabolomic Profiling of the Flavonoid Compounds in Red Wine Grapes and the Impact of Training Systems in the Southern Subtropical Region of China
by Huan Yu, Hong-Yan Li, Si-Hong Zhou, Guo Cheng, Rong-Fu Wei, Yong-Mei Zhou, Ying Zhang, Tai-Li Xie and Lan Zhang
Int. J. Mol. Sci. 2024, 25(16), 8624; https://doi.org/10.3390/ijms25168624 - 7 Aug 2024
Viewed by 278
Abstract
Flavonoids play an important role in forming wine grapes and wine quality characteristics. The flavonoids of three winter red wine grapes, Yeniang No. 2 (YN2), Marselan (Mar), and Guipu No. 6 (GP6), were analyzed by ultra-high-performance liquid chromatography–triple quadrupole mass spectrometry (UPLC-QQQ-MS). Furthermore, [...] Read more.
Flavonoids play an important role in forming wine grapes and wine quality characteristics. The flavonoids of three winter red wine grapes, Yeniang No. 2 (YN2), Marselan (Mar), and Guipu No. 6 (GP6), were analyzed by ultra-high-performance liquid chromatography–triple quadrupole mass spectrometry (UPLC-QQQ-MS). Furthermore, the flavonoids in GP6 grapevines using two types of training systems, namely, trellis (T) and espaliers (E), were also compared in this study. Overall, 196 flavonoid metabolites, including 96 flavones, 38 flavonols, 19 flavanones, 18 polyphenols, 15 anthocyanins, 7 isoflavones, and 3 proanthocyanidins, were identified. The flavonoid profiles were remarkably different among these three grape varieties, while they did not change much in the GP6 managed on trellis and espaliers. Grape varieties with different genetic backgrounds have their own unique flavonoid profiles. Compared with Mar-T, isoflavones and flavonols presented higher contents in GP6-T and YN2-T, which mainly contain glycitein, genistin, calycosin, kaempferide, isotrifoliin, and ayanin. The anthocyanin content was significantly higher in YN2-T than in the other two varieties. YN2 and GP6-T present a more stable color, with significantly more acetylated diglucosides and methylated anthocyanins in YN2-T and GP6-T than in Mar-T. Notably, GP6 had more varied flavonoids and the better characteristics to its flavonoid profile out of these three varieties, due to it containing a higher number of anthocyanins, flavone, and flavonols and the greatest number of different flavonoid metabolites (DFMs), with higher contents than YN2 and Mar. Compared with the trellis training system, the espaliers training system increased the content of flavonoids detected in GP6 grape berries; however, the composition of flavonoids strictly depends on the grape variety. Full article
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<p>(<b>A</b>) Hierarchical cluster analysis (HCA). (<b>B</b>) The numbers of all groups of flavonoids in the four treatments. (<b>C</b>) The peak area of different groups of flavonoids. (<b>D</b>) Principal component analysis (PCA). Different letters indicate significant differences among samples according to Duncan’s test (<span class="html-italic">p</span> &lt; 0.05).</p>
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<p>(<b>A</b>) Categorical metabolite statistics and (<b>B</b>) pairwise comparison of differential metabolites.</p>
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<p>(<b>A</b>) Venn diagram illustrating the overlapping and specific differential metabolites of the three comparison groups (Mar-T vs. YN2-T, Mar-T vs. GP6-T, GP6-T vs. YN2-T). (<b>B</b>) The proportions of the 17 shared metabolites in GP6-T vs. YN2-T and Mar-T vs. GP6-T. (<b>C</b>) The proportions of the 38 shared metabolites in Mar-T vs. YN2-T and GP6-T vs. YN2-T. (<b>D</b>) The proportions of the 30 shared metabolites in Mar-T vs. YN2-T and Mar-T vs. GP6-T.</p>
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<p>KEGG pathway analysis of differential metabolites in four comparison groups: (<b>A</b>) Mar-T vs. YN2-T, (<b>B</b>) Mar-T vs. GP6-T, (<b>C</b>) GP6-T vs. YN2-T, and (<b>D</b>) GP6-T vs. GP6-E.</p>
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<p>Flavonoid biosynthesis pathways in the pairwise comparisons of four treatments. The log<sub>2</sub>-transformed FPKM values were used to prepare the heatmaps and present the biological pathways connected with different metabolite expression levels.</p>
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<p>(<b>A</b>) Cyanidin, Delphinidin and Pelargonidin modified anthocyanins, (<b>B</b>) Methylated and Non-methoxyled modified anthocyanins, (<b>C</b>) Diglucosides and Monoglucoside modified anthocyanins, (<b>D</b>) Acetylated and Non-acetylated modified anthocyanins. The peak area of different modified anthocyanins among the four treatments. Different letters indicate significant differences among samples according to Duncan’s test (<span class="html-italic">p</span> &lt; 0.05).</p>
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14 pages, 1690 KiB  
Article
Enrichment of White Wine Vinegar with Aromatic Plants: The Impact on Aromatic, Polyphenolic, and Sensory Profiles
by Marin Krapac, Nikola Major, Tomislav Plavša, Ana Jeromel, Ivana Tomaz and Danijela Poljuha
Appl. Sci. 2024, 14(16), 6909; https://doi.org/10.3390/app14166909 - 7 Aug 2024
Viewed by 417
Abstract
The food industry is developing intensively, and products that, with their characteristics, enrich the food taste and aroma are widely used in the culinary arts. White wine vinegar is often used as a food condiment and as dressing in salads. This research aims [...] Read more.
The food industry is developing intensively, and products that, with their characteristics, enrich the food taste and aroma are widely used in the culinary arts. White wine vinegar is often used as a food condiment and as dressing in salads. This research aims to explore the impact of the maceration of selected aromatic plants on the organoleptic properties, bioactive compounds, and sensory profile of white wine vinegar. The plants selected for white wine (cv ‘Malvazija istarska’) vinegar aromatization were rosemary (Rosmarinus officinalis L.) and thyme (Thymus serpyllum L.) dried leaves and black elder (Sambucus nigra L. ssp. nigra) flowers (elderflowers). Vinegar flavored with rosemary had more pronounced pinewood and herbal aromas, while spicy aromas dominated the vinegar with thyme. The elderflower-flavored vinegar, on the other hand, was characterized by a floral and fruity aroma. Among the analyzed vinegars, white wine vinegar flavored with elderflower had the highest polyphenolic content. Full article
(This article belongs to the Special Issue Natural Products and Bioactive Compounds)
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<p>Scheme of vinegar production and analysis.</p>
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<p>Differences in the odorant series (herbal, floral, fruity, sweet, spicy, camphor-like, and unpleasant) of the flavored vinegars (shown as a bar graph) compared to the control treatment (shown on the <span class="html-italic">x</span>-axis) expressed as percentage changes. The odor series is represented by the sum of mean values of all compounds of the same odorant series where a significant difference between treatments was determined.</p>
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<p>Principal component analysis (PCA) for vinegars: (<b>a</b>) treatments (cases); (<b>b</b>) aromatic groups—blue letters; phenolic groups—green letters (variables). Abbreviations: Alc—alcohols, Acd—acids, Est—esters, Ter—terpenes, Otha—miscellaneous aromas, Ant—anthocyanins, Fla—flavones, FlaOL—flavonols, Fla-3-ol—flavan-3-ol, Dhf—dihydroflavonols, HbA—hydroxybenzoic acids, HcA—hydroxycinnamic acids, HppA—hydroxyphenylpropanoic acids, Stil—stilbenes, Hba—hydroxybenzaldehydes, Ty—tyrosols.</p>
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<p>Comparison of the odor properties of flavored vinegars (elderflower, rosemary, and thyme) and the control treatment. An analysis of variance (ANOVA) was performed to determine if there was a significant difference among the odor properties of the vinegars. Odor properties marked with asterisks (**, <span class="html-italic">p</span> ≤ 0.01; ***, <span class="html-italic">p</span> ≤ 0.001) show significant differences.</p>
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13 pages, 1036 KiB  
Article
Effective Ethyl Carbamate Prevention in Red Wines by Treatment with Immobilized Acid Urease
by Elisa Tavilli, Marco Esti and Marcello Fidaleo
Foods 2024, 13(16), 2476; https://doi.org/10.3390/foods13162476 - 6 Aug 2024
Viewed by 338
Abstract
Climate change poses several challenges in the wine industry, including increasing risks related to chemical food contaminants such as biogenic amines and ethyl carbamate (EC). In this work, we focused on urea removal in red wines by immobilized acid urease aiming at limiting [...] Read more.
Climate change poses several challenges in the wine industry, including increasing risks related to chemical food contaminants such as biogenic amines and ethyl carbamate (EC). In this work, we focused on urea removal in red wines by immobilized acid urease aiming at limiting EC formation during wine storage. By considering separable kinetics of catalyst deactivation and urea hydrolysis, it was possible to model the time course of urea removal in repeated uses in stirred batch reactors. Treatments based on immobilized urease of red wine enriched with 30 mg/L of urea allowed the reduction in the contaminant concentration to <5 mg/L. After 28.5 h of treatment, the observed urea level was reduced to about 0.5 mg/L, corresponding to a decrease in the potential ethyl carbamate (PEC) from 1662 μg/L to 93 μg/L, below the level of the non-enriched wine (187 μg/L). As a comparison, when treating the same wine with the free enzyme at maximum doses allowed by the EU law, urea and PEC levels decreased to only 12 mg/L and 415 μg/L respectively, after 600 h of treatment. These results show that, for red wines, urease immobilization is an effective strategy for urea removal and, thus, effective reduction in ethyl carbamate as a process contaminant. This study provides the scientific background for the future scaling-up of the process at an industrial level. Full article
(This article belongs to the Section Food Engineering and Technology)
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<p>Effect of urea concentration additions (ΔC<sub>A</sub>) on measured urea (C<sub>A</sub>, ●) and ammonia (C<sub>B</sub>, ○) concentrations in the red wine.</p>
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<p>Repeated bioconversions in the stirred batch reactor: concentrations of urea (C<sub>A</sub>, ●) and ammonia (C<sub>B</sub>, ○) as a function of time (t) (<b>a</b>); increase in ammonia concentration (ΔC<sub>B</sub>) as a function of decrease in urea concentration (−ΔC<sub>A</sub>) (□) (<b>b</b>).</p>
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<p>Effect of urea consumption and urease deactivation on the time course of urea concentration (C<sub>A</sub>). Symbols refer to the first use of the immobilized catalyst (●) or to the reuse after having been used for 24 h (○). The continuous line was obtained by fitting the data of the first and second cycles of use by using Equation (10). The square symbols (□) refer to the final urea content in the two hydrolysis batches used to determine PEC; however, their corresponding values were not used in the fitting procedure. The dashed and dash-dotted lines refer to simulations based on Equation (10) of urea hydrolysis at the hypothetical third and fourth 24 h cycles of reuse, respectively. The continuous, virtually horizontal line (✕) corresponds to urea hydrolysis occurring with the use of free urease according to the maximum dosage (375 IU/L, corresponding to 150 mg of enzyme preparation per liter of wine treated) allowed by the EU law [<a href="#B25-foods-13-02476" class="html-bibr">25</a>] assessed in a previous work on the same red wine [<a href="#B3-foods-13-02476" class="html-bibr">3</a>].</p>
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<p>Effect of free [<a href="#B3-foods-13-02476" class="html-bibr">3</a>] or immobilized urease treatments on potential ethyl carbamate (PEC) of red wine as a function of residual urea concentration (C<sub>A</sub>). The closed symbols refer to the red non-enriched wine (♦), enriched with urea at 30 mg/L (●) or enriched with urea and treated with immobilized urease for 14.5 (▲) or 28.5 h (■). The data obtained with the free enzyme refer to the red wine enriched with 30 mg/L of urea (□) and treated for 600 h at a urease dose of 375 IU/L (as set by EU law [<a href="#B25-foods-13-02476" class="html-bibr">25</a>]) (○). Bars represent standard deviations of three replicates.</p>
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12 pages, 487 KiB  
Perspective
Soil Microbial Communities and Wine Terroir: Research Gaps and Data Needs
by Gabriela Crystal Franco, Jasmine Leiva, Sanjiev Nand, Danica Marvi Lee, Michael Hajkowski, Katherine Dick, Brennan Withers, LuzMaria Soto, Benjamin-Rafael Mingoa, Michael Acholonu, Amari Hutchins, Lucy Neely and Archana Anand
Foods 2024, 13(16), 2475; https://doi.org/10.3390/foods13162475 - 6 Aug 2024
Viewed by 432
Abstract
Microbes found in soil can have a significant impact on the taste and quality of wine, also referred to as wine terroir. To date, wine terroir has been thought to be associated with the physical and chemical characteristics of the soil. However, there [...] Read more.
Microbes found in soil can have a significant impact on the taste and quality of wine, also referred to as wine terroir. To date, wine terroir has been thought to be associated with the physical and chemical characteristics of the soil. However, there is a fragmented understanding of the contribution of vineyard soil microbes to wine terroir. Additionally, vineyards can play an important role in carbon sequestration since the promotion of healthy soil and microbial communities directly impacts greenhouse gas emissions in the atmosphere. We review 24 studies that explore the role of soil microbial communities in vineyards and their influence on grapevine health, grape composition, and wine quality. Studies spanning 2015 to 2018 laid a foundation by exploring soil microbial biogeography in vineyards, vineyard management effects, and the reservoir function of soil microbes for grape-associated microbiota. On the other hand, studies spanning 2019 to 2023 appear to have a more specific and targeted approach, delving into the relationships between soil microbes and grape metabolites, the microbial distribution at different soil depths, and microbial influences on wine flavor and composition. Next, we identify research gaps and make recommendations for future work. Specifically, most of the studies utilize targeted sequencing (16S, 26S, ITS), which only reveals community composition. Utilizing high-throughput omics approaches such as shotgun sequencing (to infer function) and transcriptomics (for actual function) is vital to determining the specific mechanisms by which soil microbes influence grape chemistry. Going forward, understanding the long-term effects of vineyard management practices and climate change on soil microbiology, grapevine trunk diseases, and the role of bacteriophages in vineyard soil and wine-making would be a fruitful investigation. Overall, the studies presented shed light on the importance of soil microbiomes and their interactions with grapevines in shaping wine production. However, there are still many aspects of this complex ecosystem that require further exploration and understanding to support sustainable viticulture and enhance wine quality. Full article
(This article belongs to the Topic Advances in Analysis of Food and Beverages)
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<p>Dominant bacterial phyla in vineyard soil microbial communities.</p>
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22 pages, 5523 KiB  
Article
Elevating Skincare Science: Grape Seed Extract Encapsulation for Dermatological Care
by Maria Leonor Castro, João Azevedo-Silva, Diana Valente, Adriana Machado, Tânia Ribeiro, João Paulo Ferreira, Manuela Pintado, Oscar L. Ramos, Sandra Borges and Sara Baptista-Silva
Molecules 2024, 29(16), 3717; https://doi.org/10.3390/molecules29163717 - 6 Aug 2024
Viewed by 608
Abstract
The skin is the largest organ in the human body and serves multiple functions such as barrier protection and thermoregulation. The maintenance of its integrity and healthy structure is of paramount importance. Accordingly, technological advances in cosmetic sciences have been directed towards optimizing [...] Read more.
The skin is the largest organ in the human body and serves multiple functions such as barrier protection and thermoregulation. The maintenance of its integrity and healthy structure is of paramount importance. Accordingly, technological advances in cosmetic sciences have been directed towards optimizing these factors. Plant-derived ingredients have been explored for their bioactivity profiles and sustainable sources. Grape by-products contain a group of bioactive molecules that display important biological activities. Nonetheless, many of these molecules (e.g., phenolic compounds) are unstable and susceptible to degradation. So, their encapsulation using nano/microsystems (i.e., microdispersions) has been explored as a promising solution. In this work, two grape seed extracts were obtained, one from a single grape variety (GSE-Ov) and another from a mix of five grape varieties (GSE-Sv). These extracts were analysed for their antioxidant and antimicrobial activities, as well as their chemical composition and molecular structure. The extract that showed the most promising properties was GSE-Ov with a DPPH IC50 of 0.079 mg mL−1. This extract was encapsulated in soy lecithin microdispersions coated with pectin, with an encapsulation efficiency of 88.8%. They showed an in vitro release of polyphenols of 59.4% during 24 h. The particles displayed a zeta potential of −20.3 mV and an average diameter of 13.6 µm. Microdispersions proved to be safe under 5 and 2.5 mg mL−1 in HaCaT and HDF cell models, respectively. Additionally, they demonstrated anti-inflammatory activity against IL-1α when tested at 2 mg mL−1. This work enabled the valorisation of a by-product from the wine industry by using natural extracts in skincare products. Full article
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<p>Schematic representation of the extraction of the GSEs. Created with <a href="http://BioRender.com" target="_blank">BioRender.com</a>.</p>
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<p>Schematic representation of the encapsulation process. Created with <a href="http://BioRender.com" target="_blank">BioRender.com</a>.</p>
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<p>Chromatogram of the HPLC analysis of the GSEs and the respective identified phenolic compounds at 280 nm (blue: GSE-Sv; black: GSE-Ov).</p>
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<p>FTIR spectra of the GSEs. Red lines indicate the wavelengths of the functional groups, discussed below.</p>
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<p>Size distribution of the GSE-Ov-loaded microdispersion.</p>
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<p>SEM images of the powdered ((<b>A</b>) magnification of 4000×) and splintered ((<b>B</b>) magnification of 2000×) lyophilized GSE-Ov-loaded microdispersions.</p>
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<p>FTIR spectra of GSE-OV and the empty and GSE-OV-loaded microdispersions. Red lines indicate the wavelengths of the functional groups, discussed below.</p>
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<p>Cytotoxicity study of GSE-Ov, the GSE-Ov-loaded microdispersion, and the empty microdispersion on the HaCaT and HDF cell lines.</p>
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<p>Effect of GSE-Ov, the GSE-Ov-loaded microdispersion, and the empty microdispersion on keratinocytes (HaCaT) by assessing pro-inflammatory cytokine IL-1α and IL-6 levels under an inflammatory stimulus (pollution particles). Mean values (solid bars) are expressed as pg cytokine mg<sup>−1</sup> total protein, and standard deviations are represented by bars; * significantly different from positive control (CTR+) (<span class="html-italic">p</span> &lt; 0.05).</p>
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<p>Effect of GSE-OV, the GSE-Ov-loaded microdispersion, and the empty microdispersion on the production of pro-Collagen I α1 by HDF cells. Mean values (solid bars) are expressed as pg collagen µg<sup>−1</sup> protein and the standard deviation is represented by bars; * significantly different from the control (CTR) (<span class="html-italic">p</span> &lt; 0.05).</p>
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20 pages, 8083 KiB  
Article
Biochemical and Structural Characterization of a Novel Psychrophilic Laccase (Multicopper Oxidase) Discovered from Oenococcus oeni 229 (ENOLAB 4002)
by Isidoro Olmeda, Francisco Paredes-Martínez, Ramón Sendra, Patricia Casino, Isabel Pardo and Sergi Ferrer
Int. J. Mol. Sci. 2024, 25(15), 8521; https://doi.org/10.3390/ijms25158521 - 5 Aug 2024
Viewed by 349
Abstract
Recently, prokaryotic laccases from lactic acid bacteria (LAB), which can degrade biogenic amines, were discovered. A laccase enzyme has been cloned from Oenococcus oeni, a very important LAB in winemaking, and it has been expressed in Escherichia coli. This enzyme has [...] Read more.
Recently, prokaryotic laccases from lactic acid bacteria (LAB), which can degrade biogenic amines, were discovered. A laccase enzyme has been cloned from Oenococcus oeni, a very important LAB in winemaking, and it has been expressed in Escherichia coli. This enzyme has similar characteristics to those previously isolated from LAB as the ability to oxidize canonical substrates such as 2,2-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), 2,6-dimethoxyphenol (2,6-DMP), and potassium ferrocyanide K4[Fe(CN6)], and non-conventional substrates as biogenic amines. However, it presents some distinctiveness, the most characteristic being its psychrophilic behaviour, not seen before among these enzymes. Psychrophilic enzymes capable of efficient catalysis at low temperatures are of great interest due to their potential applications in various biotechnological processes. In this study, we report the discovery and characterization of a new psychrophilic laccase, a multicopper oxidase (MCO), from the bacterium Oenococcus oeni. The psychrophilic laccase gene, designated as LcOe 229, was identified through the genomic analysis of O. oeni, a Gram-positive bacterium commonly found in wine fermentation. The gene was successfully cloned and heterologously expressed in Escherichia coli, and the recombinant enzyme was purified to homogeneity. Biochemical characterization of the psychrophilic laccase revealed its optimal activity at low temperatures, with a peak at 10 °C. To our knowledge, this is the lowest optimum temperature described so far for laccases. Furthermore, the psychrophilic laccase demonstrated remarkable stability and activity at low pH (optimum pH 2.5 for ABTS), suggesting its potential for diverse biotechnological applications. The kinetic properties of LcOe 229 were determined, revealing a high catalytic efficiency (kcat/Km) for several substrates at low temperatures. This exceptional cold adaptation of LcOe 229 indicates its potential as a biocatalyst in cold environments or applications requiring low-temperature processes. The crystal structure of the psychrophilic laccase was determined using X-ray crystallography demonstrating structural features similar to other LAB laccases, such as an extended N-terminal and an extended C-terminal end, with the latter containing a disulphide bond. Also, the structure shows two Met residues at the entrance of the T1Cu site, common in LAB laccases, which we suggest could be involved in substrate binding, thus expanding the substrate-binding pocket for laccases. A structural comparison of LcOe 229 with Antarctic laccases has not revealed specific features assigned to cold-active laccases versus mesophilic. Thus, further investigation of this psychrophilic laccase and its engineering could lead to enhanced cold-active enzymes with improved properties for future biotechnological applications. Overall, the discovery of this novel psychrophilic laccase from O. oeni expands our understanding of cold-adapted enzymes and presents new opportunities for their industrial applications in cold environments. Full article
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<p>Multiple sequence alignments and conservation of laccases from psychrophilic, mesophilic, and thermophilic organisms is shown. Scoring of the Hmer-R1t3 (<span class="html-italic">Halomonas meridiana</span> CopA), Hpie-resA (<span class="html-italic">Halomonas piezotolerans</span>), Hcol-FME66 (<span class="html-italic">Halomonas colorata</span>), PhaMOx (<span class="html-italic">Pseudoalteromonas haloplanktis</span>), Ecol-6898 (<span class="html-italic">Escherichia coli</span> CueO), Bsub-6077 (<span class="html-italic">Bacillus subtilis</span> CotA), Ppar-3909 (<span class="html-italic">Pediococcus parvulus</span>), Llac-5298 (<span class="html-italic">Lactococcus lactis</span>), Ppen-4816 (<span class="html-italic">Pediococcus pentosaceus</span>), Lpar-4314 (<span class="html-italic">Lacticaseibacillus paracasei</span>), Lpla-J16 (<span class="html-italic">Lactiplantibacillus plantarum</span>), Paci-5930 (<span class="html-italic">Pediococcus acidilactici</span>), Lsak-LS (<span class="html-italic">Latilactobacillus sakei</span>), Ooen-2291 (<span class="html-italic">Oenococcus oeni</span> LcOe 229), and Tth-MCO (<span class="html-italic">Thermus thermophilus</span>) laccases was performed using Promals3D software (<a href="http://prodata.swmed.edu/promals3d/promals3d.php" target="_blank">http://prodata.swmed.edu/promals3d/promals3d.php</a>, accessed on 18 May 2024) [<a href="#B23-ijms-25-08521" class="html-bibr">23</a>,<a href="#B24-ijms-25-08521" class="html-bibr">24</a>]. The sequence of Paci-5930 is representative of the shortest C-terminus in LAB. The first row in each block shows conservation indices for positions with a conservation index greater than 4. The last two rows show the consensus amino acid sequence (Consensus_aa) and consensus-predicted secondary structures (Consensus_ss). Representative sequences have magenta names and they are coloured according to predicted secondary structures (red: alpha-helix, blue: beta-strand). The sequences with black names directly under a representative sequence are in the same pre-aligned group and are aligned in a fast way. Consensus predicted secondary structure symbols: alpha-helix: h; beta-strand: e. Consensus amino acid symbols are: conserved amino acids are in bold and uppercase letters; aliphatic (I, V, L): <span class="html-italic"><span style="color:red">l</span></span>; aromatic (Y, H, W, F): <span class="html-italic"><span style="color:red">@</span></span>; hydrophobic (W, F, Y, M, L, I, V, A, C, T, H): <span class="html-italic"><span style="color:red">h</span></span>; alcohol (S, T): o; polar residues (D, E, H, K, N, Q, R, S, T): p; tiny (A, G, C, S): t; small (A, G, C, S, V, N, D, T, P): s; bulky residues (E, F, I, K, L, M, Q, R, W, Y): b; positively charged (K, R, H): <b><span style="color:#00B0F0">+</span></b>; negatively charged (D, E): <b><span style="color:#00B0F0">-</span></b>; charged (D, E, K, R, H): c. Dashes indicate gaps to maximise alignment. The motifs forming the four copper ligands, which are highly conserved in laccases (conserved sequences of these motifs are HXHG, HXH, HXXHXH and HCHXXXHXXXXM/L/F), are enclosed in boxes [<a href="#B25-ijms-25-08521" class="html-bibr">25</a>].</p>
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<p>Purification and characterization of the recombinant protein LcOe 229. (<b>A</b>) PageBlue-stained 7.5% SDS-polyacrylamide gel electrophoresis of the different steps of the purification process. Lane M, page ruler unstained marker (Thermo Scientific). Lane 1: whole-cell extract from post-induction cells; lane 2: unbound proteins collected in the flowthrough; lane 3: proteins collected after washing with 5 mM imidazole buffer; lane 4: proteins collected after washing with 20 mM imidazole buffer; lanes 5–13: successive fractions of the elution from the metal-chelating chromatography on Ni<sup>2+</sup>-NTA-agarose; arrow marks recombinant protein. (<b>B</b>) Absorption spectrum of the purified protein. (<b>C</b>) Optimum pH for the oxidation of the substrates 2,2-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), 2,6-dimethoxyphenol (2,6-DMP), and potassium ferrocyanide K<sub>4</sub>[Fe(CN<sub>6</sub>)], of the LcOe 229 laccase. (<b>D</b>) Optimum temperature and thermostability of the LcOe 229 measured using ABTS as substrate. (<b>E</b>) Effect of different putative inhibitors on the laccase LcOe 229. ZnCl<sub>2</sub>: zinc chloride; Bipy.: 2,20-bipyridyl; EDTA: ethylenediaminetetraacetic acid; <span class="html-italic">Thioglyc.</span> acid: thioglycolic acid; <span class="html-italic">Cyst.</span>: cysteine HCl monohydrate; Na Az.: sodium azide; Semicarb.: semicarbazide; Rasag.: rasagiline; Phenan.: 1,10-phenanthroline; Parg.: pargyline; EDC: N-(3-dimethyl aminopropyl)-N′-ethyl carbodiimide; Depren.: deprenyl; Cyclop.: cyclopropenyl; Clorgyl.: clorgyline; NaCl: sodium chloride; NaF: sodium fluoride; control means an enzyme reaction in absence of inhibitors. The remaining activity with the different potential inhibitors is graphed as percentage respect to the control (assumed as 100% of enzyme activity). Values are means ± standard deviations of triplicate assays.</p>
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<p>Structure of LcOe 229 compared with other laccases. (<b>A</b>) Cartoon representation of LcOe 229 highlighting each domain by colour-coding (Domain 1 in cyan, Domain 2 in yellow, and Domain 3 in magenta) and the linker D2–D3 in red. (<b>B</b>) Superposition of each domain colour-coded as in (<b>A</b>) with the structure of LAB laccases Pa5930 (in blue) and Pp4816 (in orange). (<b>C</b>) Sequence and structural alignment of the C-terminal end for LcOe 229 with several LAB laccases Ppar-3909 (<span class="html-italic">P. parvulus</span>), Llac-5298 (<span class="html-italic">L. lactis</span>), Ppen-4816 (<span class="html-italic">P. pentosaceus</span>), Lpar-4314 (<span class="html-italic">L. paracasei</span>), Lpla-J16 (<span class="html-italic">L. plantarum</span>), Paci-5930 (<span class="html-italic">P. acidilactici</span>), Lsak-LS (<span class="html-italic">L. sakei</span>). Framed is a rather conserved region in LAB which presents more differences in LcOe. The scoring scheme works from 0 for the least conserved alignment position, up to 10 for the most conserved alignment position, and the identical conserved residues are indicated by asterisks (colour scale indicated on top). (<b>D</b>) Superposition of Domain 3 (in magenta) from LcOe 229 with different structures of laccases Pa5930 (in blue; PDB: 6Z0K), Pp4816 (in orange; PDB: 6XJ0), CueO (in green; PDB: 3NSD), CotA (in pink; PDB: 4Q8B), and Tth-MCO (in light brown; PDB: 2XUW). Side chains for the two Met (M348 and M390) at the entrance of T1Cu site of LcOe 229 are shown as well as the side chains of two Met at a similar site found in other laccases. For CotA, it is shown the substrate sinapic acid which lies on the Met.</p>
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<p>Organization of the copper binding sites in LcOe 229. (<b>A</b>) Representation of the configuration observed for T1Cu and TNC showing the side chain of the residues coordinating the copper ions (in orange). The His residues are color-coded as Domain 1 (in cyan) and Domain 3 (in magenta) while Cys are colored in green and Met are colored in black (sulfurs are in yellow). The two Met at the entrance of T1Cu are also shown. (<b>B</b>) Zoom of the TNC site in chain B and in chain A of LcOe 229 showing the electron density of 2fo-fc map at 1σ. The side chains of the His coordinating the coppers are color-coded as the domains. The E452 and D118 involved in proton coordination are also shown. (<b>C</b>) Structure of Histag-LcOe 229 showing how the C-terminal of one molecule (Domain 3 in magenta) is in close distance to the Met residues (M348 and M390) at the entrance of T1Cu from the other molecule (Domain 3 in transparency magenta). The central and right panel show a superposition of Histag-LcOe 220 with <span class="html-italic">M. albomyces</span> laccase (MaL) in the presence of 2,6-DMP (PDB: 3FU7) and bilirubin oxidase from <span class="html-italic">M. verrucaria</span> in the presence of ferricyanide (PDB: 6I3J). Dotted lines are an indication of distance between atoms and distance values are also shown.</p>
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17 pages, 5890 KiB  
Article
Effects of High-Hydrostatic-Pressure Treatment on Polyphenols and Volatile Aromatic Compounds in Marselan Wine
by Zicheng Yi, Danqing Zhao, Tengwen Chang, Xiang Chen, Jianrong Kai, Yang Luo, Bangzhu Peng, Binkun Yang and Qian Ge
Foods 2024, 13(15), 2468; https://doi.org/10.3390/foods13152468 - 5 Aug 2024
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Abstract
This study investigated the effects of high-hydrostatic-pressure (HHP) treatment of varying intensity (100–600 MPa) and duration (10–30 min) on polyphenols and volatile aromatic compounds in Marselan red wine. The types and concentrations of polyphenols and volatile aromatic compounds were compared before and after [...] Read more.
This study investigated the effects of high-hydrostatic-pressure (HHP) treatment of varying intensity (100–600 MPa) and duration (10–30 min) on polyphenols and volatile aromatic compounds in Marselan red wine. The types and concentrations of polyphenols and volatile aromatic compounds were compared before and after HHP treatment; the results indicated that HHP treatment at 300 MPa for 20 min significantly increased the total polyphenol content to 369.70 mg/L, a rise of 35.82%. The contents of key polyphenols, such as resveratrol and protocatechuic acid, were significantly enhanced. Furthermore, while the total content of volatile aromatic compounds did not change significantly under this condition compared to the untreated samples, the concentration of ester compounds significantly increased to 1.81 times that of the untreated group, thereby enriching the floral and fruity aromas of the wine and effectively improving its aromatic profile and sensory quality. Principal component analysis (PCA) further validated the positive impact of HHP treatment on the flavor characteristics of Marselan red wine. These findings provide technical support for the use of HHP in improving wine quality. Full article
(This article belongs to the Special Issue The Past, Present and Future of the Fruit Wine Industry)
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<p>The effect of different HHP treatments on polyphenol content. As the treatment duration remains constant (10 min, 20 min, 30 min), polyphenolic compounds vary with changes in treatment pressure.</p>
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<p>The effects of different HHP treatments on the content of polyphenolic substances. Changes in polyphenolic compounds in wine as the processing time varies with constant pressure (100 MPa–600 MPa).</p>
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<p>PCA plot of polyphenolic substances under different HHP treatments.</p>
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<p>Distribution histogram of aroma substance content in wine samples treated with different HHP processes: (<b>a</b>) changes in volatile aroma substances under different HHP treatments; (<b>b</b>) changes in ester-type volatile aroma substances under different HHP treatments.</p>
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<p>Effects of different HHP treatments on volatile aroma compounds with OAV &gt; 1. As the treatment duration remains constant (10 min, 20 min, 30 min), volatile aroma compounds vary with changes in treatment pressure.</p>
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<p>Effects of different HHP treatments on volatile aroma compounds with OAV &gt; 1. Changes in volatile aroma compounds in wine as processing time varies with constant pressure (100 MPa–600 MPa).</p>
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<p>PCA plot of volatile aromatic substances with OAV &gt; 1 under different HHP treatments.</p>
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<p>Average sensory scores of wine samples under Different HHP treatments: (<b>a</b>) appearance and taste; (<b>b</b>) aroma.</p>
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15 pages, 1113 KiB  
Article
Fermentation Performances and Aroma Contributions of Selected Non-Saccharomyces Yeasts for Cherry Wine Production
by Federico Bianchi, Michele Avesani, Marilinda Lorenzini, Giacomo Zapparoli and Barbara Simonato
Foods 2024, 13(15), 2455; https://doi.org/10.3390/foods13152455 - 3 Aug 2024
Viewed by 392
Abstract
This study evaluates the fermentation performances of non-Saccharomyces strains in fermenting cherry must from Italian cherries unsuitable for selling and not intended to be consumed fresh, and their effects on the chemical composition of the resulting wine. Fermentation trials in 100 and [...] Read more.
This study evaluates the fermentation performances of non-Saccharomyces strains in fermenting cherry must from Italian cherries unsuitable for selling and not intended to be consumed fresh, and their effects on the chemical composition of the resulting wine. Fermentation trials in 100 and 500 mL of must were carried out to select 21 strains belonging to 11 non-Saccharomyces species. Cherry wines obtained by six select strains were chemically analyzed for fixed and volatile compounds. Quantitative data were statistically analyzed by agglomerative hierarchical clustering, partial least squared discriminant analysis, and principal component analysis. Wines revealed significant differences in their composition. Lactic acid and phenylethyl acetate levels were very high in wines produced by Lachancea and Hanseniaspora, respectively. Compared to S. cerevisiae wine, non-Saccharomyces wines had a lower content of fatty acid ethyl esters 4-vinyl guaiacol and 4-vinyl phenol. The multivariate analysis discriminated between wines, demonstrating the different contributions of each strain to aroma components. Specifically, all wines from non-Saccharomyces strains were kept strictly separate from the control wine. This study provided comprehensive characterization traits for non-conventional strains that enhance the aroma complexity of cherry-based wine. The use of these yeasts in cherry wine production appears promising. Further investigation is required to ascertain their suitability for larger-scale fermentation. Full article
(This article belongs to the Section Drinks and Liquid Nutrition)
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<p>Kinetic of CO<sub>2</sub> production (% <span class="html-italic">w</span>/<span class="html-italic">v</span>) during the fermentation of pasteurized cherry must A by 10 selected yeasts and <span class="html-italic">S. cerevisiae</span> EC1118<sup>®</sup>. (<b>A</b>) Kinetics of CO<sub>2</sub> production of strains <span class="html-italic">S. cerevisiae</span> EC1118, <span class="html-italic">H. vineae</span> Hv1, <span class="html-italic">H. vineae</span> Hv20, <span class="html-italic">H. osmophila</span> Ho22, and <span class="html-italic">H. osmophila</span> Ho3; (<b>B</b>) Kinetics of CO<sub>2</sub> production of strains <span class="html-italic">St. bacillaris</span> Stb7, and <span class="html-italic">S. bacillaris</span> Stb91; (<b>C</b>) Kinetics of CO<sub>2</sub> production of strains <span class="html-italic">L. fermentati</span> Lf2, and <span class="html-italic">T. delbrueckii</span> Td7; (<b>D</b>) Kinetics of CO<sub>2</sub> production of strains <span class="html-italic">Z. bailii</span> Zb19, and <span class="html-italic">Z. bailii</span> Zb23.</p>
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<p>Heat map with dendrograms made with AHC corresponding to aroma compounds of cherry-based wines produced by <span class="html-italic">S. cerevisiae</span> EC1118<sup>®</sup> (control), <span class="html-italic">T. delbrueckii</span> Td7 (TD), <span class="html-italic">H. vineae</span> Hv20 (HV), <span class="html-italic">H. osmophila</span> Ho3 (HO), <span class="html-italic">L. fermentati</span> Lf2 (LF), <span class="html-italic">St. bacillaris</span> Stb91 (STB), and <span class="html-italic">Z. bailii</span> Zb19 (ZB). The relative content of each molecule is illustrated through a color scale from white (minimum) to dark red (maximum).</p>
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<p>VIP score (&gt;1) and relative content (color scale from white as minimum to red as maximum) of aroma compounds of cherry-based wines produced by <span class="html-italic">S. cerevisiae</span> EC1118<sup>®</sup> (control), <span class="html-italic">T. delbrueckii</span> Td7 (TD), <span class="html-italic">H. vineae</span> Hv20 (HV), <span class="html-italic">H. osmophila</span> Ho3 (HO), <span class="html-italic">L. fermentati</span> Lf2 (LF), <span class="html-italic">St. bacillaris</span> Stb91 (STB), and <span class="html-italic">Z. bailii</span> Zb19 (ZB) obtained by PLS-DA analysis.</p>
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