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Keywords = Saccharomyces cerevisiae Fm17

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16 pages, 2303 KB  
Article
Cytotoxicity of Quantum Dots in Receptor-Mediated Endocytic and Pinocytic Pathways in Yeast
by Onyinye Okafor and Kyoungtae Kim
Int. J. Mol. Sci. 2024, 25(9), 4714; https://doi.org/10.3390/ijms25094714 - 26 Apr 2024
Cited by 3 | Viewed by 1551
Abstract
Despite the promising applications of the use of quantum dots (QDs) in the biomedical field, the long-lasting effects of QDs on the cell remain poorly understood. To comprehend the mechanisms underlying the toxic effects of QDs in yeast, we characterized defects associated with [...] Read more.
Despite the promising applications of the use of quantum dots (QDs) in the biomedical field, the long-lasting effects of QDs on the cell remain poorly understood. To comprehend the mechanisms underlying the toxic effects of QDs in yeast, we characterized defects associated with receptor-mediated endocytosis (RME) as well as pinocytosis using Saccharomyces cerevisiae as a model in the presence of cadmium selenide/zinc sulfide (CdSe/ZnS) QDs. Our findings revealed that QDs led to an inefficient RME at the early, intermediate, and late stages of endocytic patch maturation at the endocytic site, with the prolonged lifespan of GFP fused yeast fimbrin (Sac6-GFP), a late marker of endocytosis. The transit of FM1-43, a lipophilic dye from the plasma membrane to the vacuole, was severely retarded in the presence of QDs. Finally, QDs caused an accumulation of monomeric red fluorescent protein fused carbamoyl phosphate synthetase 1 (mRFP-Cps1), a vacuolar lumen marker in the vacuole. In summary, the present study provides novel insights into the possible impact of CdSe/ZnS QDs on the endocytic machinery, enabling a deeper comprehension of QD toxicity. Full article
(This article belongs to the Section Molecular Nanoscience)
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18 pages, 1478 KB  
Article
Exploitation of Cocoa Pod Residues for the Production of Antioxidants, Polyhydroxyalkanoates, and Ethanol
by Licelander Hennessey Ramos, Miluska Cisneros-Yupanqui, Diana Vanessa Santisteban Soto, Anna Lante, Lorenzo Favaro, Sergio Casella and Marina Basaglia
Fermentation 2023, 9(9), 843; https://doi.org/10.3390/fermentation9090843 - 14 Sep 2023
Cited by 18 | Viewed by 5222
Abstract
Cocoa pod husks (CPH) and cocoa bean shells (CBS) are the main by-products of the cocoa industry and a source of bioactive compounds. These residues are not completely used and thrown in the fields without any treatment, causing environmental problems. Looking for a [...] Read more.
Cocoa pod husks (CPH) and cocoa bean shells (CBS) are the main by-products of the cocoa industry and a source of bioactive compounds. These residues are not completely used and thrown in the fields without any treatment, causing environmental problems. Looking for a holistic valorization, the aim of this work was first to deeply characterize CPH and CBS in their chemical composition, amino acid, and fatty acid profiles, as well as their application as antioxidants. CBS had a high level of protein (17.98% DM) and lipids (16.24% DM) compared with CPH (4.79 and 0.35% DM respectively). Glutamic acid and aspartic acid were the predominant amino acids. The total phenolic compounds (TPC) detected in the ethanolic extracts of CPH and CBS were similar to pyrogallol as the main detected polyphenol (72.57 mg/L). CBS ethanolic extract showed a higher antioxidant activity than CPH. Both extracts increased the oxidation stability of soybean oil by 48% (CPH) and 32% (CBS). In addition, alkaline pretreatment of CPH was found suitable for the release of 15.52 ± 0.78 g glucose/L after subsequent saccharification with the commercial enzyme Cellic®. CTec2. Alkaline hydrolyzed and saccharified CPH (Ahs-CPH) was assessed for the first time to obtain polyhydroxy alkanoate (PHAs) and bioethanol. Ahs-CPH allowed the growth of both Cupriavidus necator DSM 545 and Saccharomyces cerevisiae Fm17, well-known as PHA- and bioethanol-producing microbes, respectively. The obtained results suggest that such agricultural wastes have interesting characteristics with new potential industrial uses that could be a better alternative for the utilization of biomass generated as million tons of waste annually. Full article
(This article belongs to the Special Issue Sustainable Development of Food Waste Biorefineries)
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15 pages, 3218 KB  
Article
Constitutive High Expression Level of a Synthetic Deleted Encoding Gene of Talaromyces minioluteus Endodextranase Variant (rTmDEX49A–ΔSP–ΔN30) in Komagataella phaffii (Pichia pastoris)
by Roberto Carlos Arísticas Ribalta, Lisandra Martínez Valdés, Meinardo Lafargue Gámez, Sonia Rodríguez Davydenko, Eric Dubreucq, Veronique Perrier, Benoît Moreau and Reinaldo Fraga Vidal
Appl. Sci. 2022, 12(15), 7562; https://doi.org/10.3390/app12157562 - 27 Jul 2022
Cited by 1 | Viewed by 2903
Abstract
In the sugar industry, dextran generates difficulties in the manufacturing process. Using crude dextranase (EC 3.2.1.11) to eliminate dextran in sugar is an effective practice. In this study, a synthetic dextranase-encoding gene of the filamentous fungus Talaromyces minioluteus, lacking its putative native [...] Read more.
In the sugar industry, dextran generates difficulties in the manufacturing process. Using crude dextranase (EC 3.2.1.11) to eliminate dextran in sugar is an effective practice. In this study, a synthetic dextranase-encoding gene of the filamentous fungus Talaromyces minioluteus, lacking its putative native signal peptide (1–20 amino acids) and the next 30 amino acids (rTmDEX49A–ΔSP–ΔN30), was fused to the Saccharomyces cerevisiae prepro α–factor (MFα–2) signal sequence and expressed in Komagataella phaffii under the constitutive GAP promoter. K. phaffii DEX49A–ΔSP–ΔN30, constitutively producing and secreting the truncated dextranase, was obtained. The specific activity of the truncated variant resulted in being nearly the same in relation to the full-length mature enzyme (900–1000 U·mg−1 of protein). At shaker scale (100 mL) in a YPG medium, the enzymatic activity was 273 U·mL−1. The highest production level was achieved in a fed-batch culture (30 h) at 5 L fermenter scale using the FM21–PTM1 culture medium. The enzymatic activity in the culture supernatant reached 1614 U·mL−1, and the productivity was 53,800 U·L−1·h−1 (53.8 mg·L−1·h−1), the highest reported thus far for a DEX49A variant. Dextran decreased rTmDEX49A–ΔSP–ΔN30 mobility in affinity gel electrophoresis, providing evidence of carbohydrate–protein interactions. K. phaffii DEX49A–ΔSP–ΔN30 shows great potential as a methanol-free, commercial dextranase production system. Full article
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