Search Results (72)

In this study, a comprehensive derivatization strategy for biogenic amines based on the formation of volatile carbamate derivatives using 2,2,2-trifluoroethyl chloroformate (TFECF) was successfully developed and evaluated. A series of biogenic amine derivatives was obtained in excellent yields (94–99%) and structurally confirmed using NMR, MS, and crystal structure analysis. The reagent demonstrated high reactivity toward primary and secondary amines, providing derivatives of excellent purity and satisfactory volatility. The applicability of the proposed procedure to real food matrices was demonstrated using GC-MS. The obtained results were compared with the corresponding ethyl chloroformate (ECF) derivatives. TFECF derivatives exhibited significantly improved volatility, reflected in shorter retention times and enhanced analytical performance. Full article

(This paper presents the synthesis of a novel copper(II) metal complex with ethyl (2-(methylcarbamoyl)phenyl)carbamate and 3-methylquinazoline-2,4(1H,3H)-dione. The characterization of the compound was conducted through various techniques, including melting point determination, microwave plasma atomic emission spectrometry (MP-AES) for Cu, attenuated total reflection (ATR), IR, ¹H NMR, and ¹³C NMR spectroscopy. The coordination compound was obtained after mixing water solutions of the metal salt and the ligand dissolved in DMSO and water solutions of NaOH, in a metal-to-ligand-to-base ratio of 1:2:2. The ligand and the metal chloride were brought into the reaction at room temperature in DMSO and H₂O as solvents, respectively. The results indicate the successful formation of a stable mixed-ligand Cu(II) coordination compound involving N,O-donor ligands. Based on the obtained data, we assumed that the ligands are coordinated through N- and O-donor atoms. Spectroscopic data suggested that the ligand (3-methylquinazoline-2,4(1H,3H)-dione), by using (NaOH), coordinated to a metal ion as a monodentate ligand through the nitrogen atom of the NH group and ethyl (2-(methylcarbamoyl)phenyl)carbamate coordinated in a bidentate fashion through the N- and O-donor atoms of ester group. Additionally, two hydroxyl groups were bridged for two metal ions into the formed dimer structure. Full article

Ethyl carbamate, a genotoxic chemical contaminant present in fermented alcoholic beverages and foods, is formed from naturally occurring substances in these beverages and foods. Studies have shown that the content of ethyl carbamate can increase significantly during product storage and maturation, especially if favorable conditions are present. Higher levels of ethyl carbamate have been associated with distilled alcoholic beverages, mainly obtained from stone fruits. Ethyl carbamate content is lower in fermented foods, such as bread, yogurt, and fermented sauces. EC formation occurs through several different pathways in food systems. A primary pathway involves select compounds reacting with ethanol (EtOH); therefore, the majority of the research has focused on the occurrence of EC in alcoholic beverages Due to health risks, some countries have imposed legal limits on carbamate content in alcoholic beverages. Full article

In response to the growing interest in less conventional alcoholic beverages, this study aimed to identify novel yeast strains suitable for sake production, with a focus on their potential application in bioflavouring. Commercially available strains of bottom-fermenting brewing yeasts (Saccharomyces pastorianus), a cryotolerant wine yeast (Saccharomyces bayanus), and a wild wine yeast (Torulaspora delbrueckii) were evaluated. The quality characteristics of sake obtained using non-conventional yeasts were compared with sake produced using Saccharomyces cerevisiae K7, one of the most commonly used strains in sake brewing. Sake made with non-conventional yeasts exhibited differences in fermentation kinetics, chemical composition, and sensory properties. Wine yeasts produced sake with the most favorable ester profile, markedly distinct from those obtained with other yeast strains used in the study. Compared to the conventional strain, the concentrations of the key contributors to the fruity/floral aroma, namely 3-methylbutyl acetate and ethyl hexanoate, in sake produced with S. bayanus were higher by 249.5% and 199.3%, respectively. The wine yeast S. bayanus may be considered the most promising strain for sake production due to its ability to generate elevated levels of volatile aroma compounds associated with Ginjo-ka characteristics, as well as its effectiveness in supporting a consistent and efficient alcoholic fermentation process. Full article

The aim of this paper is to obtain ethyl (2-(methylcarbamoyl)phenyl)carbamate and its metal complexes as promising antimicrobial agents. The title compound was synthesized using the ring-opening of isatoic anhydride with methylamine and further acylation with ethyl chloroformate. All metal complexes were successfully obtained after mixing the ligand dissolved in DMSO and water solutions of the corresponding metal salts and sodium hydroxide, in a metal-to-ligand-to base ratio 1:2:2. As a result, mixed ligand complexes of ethyl 2-(methylcarbamoyl)phenyl)carbamate and 3-methylquinazoline-2,4(1H,3H)-dione were obtained. The obtained complexes were characterized by their melting points, FTIR, NMR spectroscopy, and MP-AES. Then, the antimicrobial effect of the compounds against both Gram-negative and Gram-positive bacteria, yeasts, and fungi was studied. Only the Co(II) complex showed antimicrobial activity against almost all Gram-positive and Gram-negative bacteria. The cobalt complex exhibited promising antimicrobial activity against Gram-positive Micrococcus luteus with inhibition zones of 20 mm, Listeria monocytogenes (15 mm), Staphylococcus aureus (13 mm), as well as Gram-negative Klebsiella pneumoniae (13 mm) and Proteus vulgaris (13 mm). Given the potential of metal complexes as antimicrobial agents, understanding their cytotoxic effects is crucial for evaluating their therapeutic safety. To assess the in vitro biocompatibility of the experimental compounds, a range of cell viability assays was conducted using human malignant leukemic cell lines (LAMA-84, K-562) and normal murine fibroblast cells (CCL-1). The Ni(II) complex shows IC₅₀ = 105.1 µM against human malignant leukemic cell lines LAMA-84. Based on the reported results, it may be concluded that the mixed cobalt complex of 2-(methylcarbamoyl)phenyl)carbamate and 3-methylquinazoline-2,4(1H,3H)-dione can be attributed as a promising antimicrobial agent. Future in vivo tests will contribute to establishing the antimicrobial properties of this complex. Full article

The relationship between protein structure and function is intrinsically interconnected, as the structure of a protein directly determines its functional properties. To investigate the effects of temperature and pressure on protein function, this study employed ethyl carbamate (EC) hydrolase as a model food enzyme and conducted molecular dynamics (MD) simulations under varying temperature and pressure levels to elucidate its structure–function relationship. By systematically analyzing the dynamic changes in root mean square deviation (RMSD), root mean square fluctuation (RMSF), radius of gyration (Rg), solvent accessible surface area (SASA), hydrogen bonding, catalytic pocket conformation, and packing density under different temperature and pressure conditions, we revealed the structural adaptability of EC hydrolase. Furthermore, we analyzed the characteristics of EC hydrolase using molecular dynamics simulations with temperature and pressure levels, as well as conformational bias-based computer-aided engineering, providing both theoretical and experimental foundation for the adaptability mechanisms of enzymes under extreme conditions. Full article

Malolactic fermentation (MLF), an essential enological process for wine deacidification and aroma development, is predominantly mediated by Oenococcus oeni (O. oeni). This investigation characterized 170 indigenous O. oeni isolates from two principal Chinese viticultural regions (Yinchuan, Ningxia, and Changli, Hebei) through polyphasic analysis. Forty-nine strains demonstrating genetic potential for efficient malate metabolism and biosafety compliance (absence of ethyl carbamate and biogenic amines genes) were subjected to adaptive laboratory evolution under enologically relevant stress conditions. Comparative evaluation with the superior indigenous strain SD-2a revealed eight stress-adapted isolates exhibiting superior MLF kinetics, completing L-malic acid degradation in Marselan wine. Solid-phase microextraction-gas chromatography-mass spectrometry (SPME-GC-MS) profiling identified three isolates’ (3-31, 9-10, and 9-50) significant enhancement of key fermentation aromas in experimental fermentations. These oenologically adapted indigenous strains demonstrate promising potential as regional-specific starter cultures, providing a scientific foundation for developing terroir-expressive winemaking practices and optimizing microbial resources in China’s wine industry. Full article

Ethyl carbamate (EC), known as urethane, is a naturally occurring potentially carcinogenic metabolite that is widely found in alcoholic beverages and other food-related fermented products. The concern related to the presence of the EC and its toxicity in regularly consumed fermented alcoholic beverages raises global interest in assessing the possible risks to human health. EC mitigation approaches, such as molecular imprinting technology (MIT), have been proposed to target EC while preserving the sensory quality of fermented alcoholic beverages. This review explores the principles of MIT, the advantages and disadvantages of the most common polymerisation approach for molecularly imprinted polymer (MIP) synthesis, the analytical techniques used for MIP characterisation, and the strategies used to mitigate EC in fermented alcoholic beverages, with studies reporting removal efficiencies of up to 84%. Additionally, it highlights the novelty and potential of MIPs, offering practical insights into their integration within the production of fermented alcoholic beverages, highlighting their scalability and cost-effectiveness compared to traditional EC mitigation strategies. Full article

Background/Objectives: Natural cytokinins are a promising group of anti-tumor agents. In this work, we hypothesized that modification of the ethylenediurea moiety with alkyl and oxygen-containing groups could be a way to enhance the anti-proliferative properties of the molecule. Methods: Ten new analogs of ethylenediurea with these substitutions were tested for anti-proliferative activity in the human cancer cell lines MDA-MB-231 (breast cancer), A-375 (melanoma), and U-87 MG (glioblastoma) during 72 h of incubation using resazurin test and evaluated the substances receptor using molecular docking. Results: The compound with the carbamate link and ethylene substituent on the phenyl ring inhibited proliferation in these models by 70–90% without cytotoxic effects. The compound did not affect the viability of the immortalized fibroblast cell line Bj-5ta. The compound was also able to enhance the action of doxorubicin and temozolomide by about 20%. According to the molecular modeling data, the probable receptor target for the synthesized compound was the A2AR adenosine receptor. Conclusions: The results obtained on the ethylenediurea analogs with ethyl substituent in the aromatic ring are promising for the development of novel anticancer therapeutics. Full article

Lipid-like nanoparticles (LLNPs) have been shown to be an effective encapsulation and delivery tool for therapeutic molecules. While the preclinical development of lipid nanoparticle formulations has been of paramount importance, next-generation LLNPs present an opportunity of enhanced biocompatibility. With the change in amido functionality as part of the core backbone, our target, carbamate functionality within the LLNP core scaffold, was realized upon reaction of a protected amino alcohol onto the isocyanate generated in situ via a Curtius rearrangement. The single-step assembly of carbamate functionality starting from cyclohexane carboxylic acid in the presence of diphenylphosphoryl azide (DPPA) exceeds the metrics set forth for the rapid installment and enhanced biodegradability of next-generation lipid-like nanoparticles. Full article

In winemaking, malolactic fermentation (MLF), which converts L-malic acid to L-lactic acid, is often applied after the alcoholic fermentation stage to improve the sensory properties of the wine and its microbiological stability. MLF is usually performed by lactic acid bacteria, which, however, are sensitive to the conditions of alcoholic fermentation. Therefore, the development of wine yeast strains capable of both alcoholic fermentation and MLF is an important task. Using genome editing, we engineered a modified variant of the triploid wine yeast strain Saccharomyces cerevisiae I-328, in which the CAR1 arginase gene was replaced by the malate permease gene from Schizosaccharomyces pombe and the malolactic enzyme gene from Oenococcus oeni. Genome-wide transcriptional profiling confirmed the expression of the introduced genes and revealed a limited effect of the modification on global gene expression. Winemaking experiments show that genome editing did not affect fermentation activity and ethanol production, while use of the modified strain resulted in a tenfold reduction in malate content with simultaneous formation of lactate. The resulting wines had a softer and more harmonious taste compared to wine obtained using the parental strain. Inactivation of arginase, which forms urea and L-ornithine through the breakdown of arginine, also resulted in a twofold decrease in the content of urea and the carcinogenic ethyl carbamate in wine. Thus, the new strain with the replacement of the arginase gene with the MLF gene cassette is promising for use in winemaking. Full article

Enzymatic degradation of the carcinogen ethyl carbamate (EC) and its precursor urea is a promising method for controlling EC levels in alcoholic beverages. However, limited enzymes with EC-hydrolyzing activity and low ethanol or acid tolerance hinder their practical application. Here, a new urease with urea- and EC-hydrolyzing activities from Providencia sp. LBBE was characterized. The enzyme displayed considerable ethanol tolerance, retaining 42.4% activity after 1 h of incubation with 30% (v/v) ethanol at 37 °C. It exhibited broad pH tolerance (pH 3.0–8.0), with optimal pH 7.0 for EC and 7.5 for urea. After treatment at pH 4.5 and 5.0, it retained 41.3% and 59.4% activity, respectively. The K_m and V_max for EC and urea at pH 4.5 were 515.6 mM, 33.9 µmol/(min⸱mg) and 32.0 mM, 263.6 µmol/(min⸱mg), respectively. Using 6000 U/L purified enzyme at 30 °C for 9 h, 49.8% and 81.6% of urea was removed from rice wine (pH 4.5 and 7.0), respectively. No appreciable reduction in EC was observed under identical conditions, which may be ascribed to the minimal EC affinity. This study contributes to the future realization of the effective control of EC content in alcoholic beverages. Full article

Fermented foods have gained global attention for their unique flavor and immense health benefits. These flavor compounds and nutrients result from the metabolic activities of microorganism during fermentation. However, some unpleasant sensory characteristics and biohazard substances could also be generated in fermentation process. These quality and safety issues in fermented foods could be addressed by endogenous enzymes. In this review, the applications of enzymes in quality control of fermented foods, including texture improvement, appearance stability, aroma enhancement, and debittering, are discussed. Furthermore, the enzymes employed in eliminating biohazard compounds such as ethyl carbamate, biogenic amines, and nitrites, formed during fermentation, are reviewed. Advanced biological methods used for enhancing the enzymatic activity and stability are also summarized. This review focused on the applications and future prospects of enzymes in the improvement quality and safety qualities of fermented foods. Full article

In the present study, changes in the physicochemical indices, ethyl carbamate (EC) precursor and EC contents, and microbial communities of fermented grains under different fermentation patterns during strong-aroma Baijiu (SAB) fermentation and changes in EC precursor and EC contents during distillation were investigated to study EC formation during these processes. In detail, the amounts of sorghum added in protocols C and D were half those added in protocols A and B (the normal SAB-producing technology). When fermented for about 30 to 35 days, the fermented grains of protocols A and C were, respectively, remixed with Daqu and second-distilled SAB (so-called “Huijiu jiaqu”, HJJQ) and fermented for about 30 to 40 days. The results showed that the acidities of the final fermented grains of protocols A (2.43 ± 0.09 mmol/10 g) and C (3.18 ± 0.08 mmol/10 g) were lower than those of protocols B (3.71 ± 0.07 mmol/10 g) and D (4.66 ± 0.10 mmol/10 g), while the alcohol contents in the final fermented grains of protocols A (18.33 ± 0.76%) and C (15.33 ± 1.08%) were higher than those of protocols B (5.10 ± 0.85%) and D (1.85 ± 0.62%). No significant differences were observed in the other physicochemical indices among the samples. The HJJQ operation significantly increases the alcohol content and reduces the acidity of the fermented grains but has little influence on the other physicochemical indices during SAB fermentation. Excluding the influence of the HJJQ operation and a half input of sorghum on the EC precursor and EC contents for the fermented grains of protocol B, the linear relationships between the EC content and alcohol (R²: 0.4465), citrulline (R²: 0.6962), urea (R²: 0.4705), and HCN (R²: 0.6324) contents were good (all the confidence levels were at 0.05), indicating that these compounds were the dominant EC precursors during SAB fermentation. HJJQ also facilitated the reaction between alcohol and other EC precursors, decreasing EC precursor content and increasing the EC content. KEGG pathway analysis demonstrated that EC precursors were mainly synthesized by alcohol and arginine metabolism. HCN (R²: 0.3875 to 0.8198) and alcohol (R²: 0.4642 to 0.8423) were the dominant EC precursors during SAB distillation. Overall, the HJJQ operation, especially in protocol C, could significantly reduce the content of EC in base SAB, and the base SAB obtained was of good quality. This, therefore, may be an alternative and effective way to reduce the EC content in base Baijiu. Full article

Chemometric evaluation of potentially harmful volatile compound and toxic metal(loid) distribution patterns in fruit spirits relating to distinct fruit classes most commonly used in spirit production highlighted the potential of several volatiles as candidates for differentiation markers while dismissing toxic metal(loid)s. Pome fruit and grape pomace spirits were mostly characterized by a lower abundance of n-propanol, methanol, ethyl acetate and acetaldehyde, while stone fruit spirits contained lower amounts of isoamyl alcohol and isobutanol. Chemometric analysis of the fruit spirit composition of aromatics identified additional potential markers characteristic for certain fruits—benzoic acid ethyl ester, benzyl alcohol, benzaldehyde, butanoic acid 3-methyl-ethyl ester, butanoic acid 2-methyl-ethyl ester and furfural. This study explored the variability in the risk potential of the investigated spirits, considering that some chemicals known to be detected in spirits are potent health hazards. Ethyl carbamate in combination with acetaldehyde showed a higher potential risk in stone fruit spirits, methanol in stone and pome fruit spirits and acetaldehyde in grape pomace spirits. It is of great interest to evaluate to what extent consumers’ preference for spirits of distinct fruit types affects health risks. Consumers of stone fruit spirits are potentially at higher risk than those consuming pome fruit or grape pomace spirits. Full article