Search Results (11)

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

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

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

Ethyl carbamate (EC) is a 2A classified carcinogen in Chinese liquor that has raised many problems regarding food safety. Applying microorganisms to control the content of EC precursors in fermented grains has been proven as an effective method to reduce EC in alcoholic beverages. However, the utilization of microorganisms to decrease the precursors of EC (urea and cyanide) is still incomplete in regard to Chinese liquor. Thus, it is necessary to isolate strains with the degradative activities of urea and cyanide. Herein, Bacillus sonorensis F3 and Bacillus licheniformis YA2 strains were isolated from the fermented grains through multiple rounds of high-throughput screening, and the degradative abilities in urea and cyanide reached 95.72% and 75.48%, respectively. In addition, the urease from the B. sonorensis F3 strain and the carbon nitrogen hydrolase from the B. licheniformis YA2 strain were identified by the heterogeneous expression in Escherichia coli. Then, both F3 and YA2 strains were combined at a ratio of 5:1 and applied to eliminate the EC in the simulated fermentation of Chinese liquor; as a result, 51.10% of EC was reduced without affecting the main composition of flavor substances. The obtained strains have great potential in terms of the improvement of quality and safety of Chinese liquor. Full article

Ethyl carbamate, the ethyl ester of carbamic acid, has been identified in fermented foods and alcoholic beverages. Since ethyl carbamate is a probable human carcinogen, reduction of its content is important for food safety and human health. In alcoholic beverages, ethyl carbamate is mostly formed from the reaction of ethanol with urea, citrulline and carbamyl phosphate during fermentation and storage. These precursors are generated from arginine metabolism by wine yeasts and lactic acid bacteria. This review summarizes the mechanisms of ethyl carbamate formation, its impact on human health and methods used in winemaking to minimize its content. These approaches include genetic modification of Saccharomyces cerevisiae wine strains targeting pathways of arginine transport and metabolism, the use of lactic acid bacteria to consume arginine, direct degradation of ethyl carbamate by enzymes and microorganisms, and different technological methods of grape cultivation, alcoholic fermentation, wine aging, temperature and duration of storage and transportation. Full article

Baijiu is a popular alcoholic beverage with a long history in China. However, the widespread presence of the ethyl carbamate (EC) carcinogen has raised many food safety concerns. To date, the main precursors of EC and its formation process have not been determined, resulting in difficulty controlling EC in Baijiu. In this study, the main precursors of EC are identified as urea and cyanide during the process of brewing for different flavors of Baijiu, while the dominant stage in which EC formation occurs is during the process of distillation rather than fermentation. In addition, the effects of temperature, pH value, alcohol concentration and metal ions on the formation of EC are confirmed. In the following study, the main precursor of EC is identified as cyanide during the process of distillation, and a combination of optimizing the distillation device and adding copper wire is proposed. Furthermore, the effect of this novel strategy is examined in gaseous reactions between cyanide and ethanol, reducing the concentration of EC by 74.0%. Finally, the feasibility of this strategy is verified in simulated distillations of fermented grains, reducing the formation of EC by 33.7–50.2%. This strategy has great application potential in industrial production. Full article

Background: Ethyl carbamate (EC), a byproduct that naturally forms in fermented foods, can cause tumors and cell death and is classified as a probable human carcinogen (Group 2A). EC is naturally formed through the alcoholysis reaction between ethanol and carbamyl compounds. The major precursors and dominantly emerging stages of EC differ with disparate food types, including soy sauce. This work aimed to clarify the formation of EC and its influence factors throughout the soy sauce production process with or without high-pressure process (HPP) treatment. Methods: Tetragenococcus halophilus, Pediococcus acidilactici, Zygosaccharomyces rouxii, and Candida versatilis were added to soy sauce. The levels of citrulline and EC were measured, and a 16S and ITS assay investigated the microbiota. Results: L-citrulline production was found in each group after fermentation for one month. In addition, L-citrulline levels were generated the most in group D (500 MPa treated raw soy sauce with 12% saltwater and mixed fermentation bacteria, including T. halophilus,P. acidilactici,Z. rouxii, and C. versatilis) and group E (soy sauce fermentation with 12% saltwater without HPP treatment) compared to group F (soy sauce fermentation with 18% saltwater without HPP treatment). Conclusions: These results indicated that salt concentration and mixed fermentation bacteria (T. halophilus,P. acidilactici,Z. rouxii,C. versatilis) might not be major factors for L-citrulline production. Full article

Ethyl carbamate (EC) is a potential carcinogen that forms spontaneously during Chinese rice wine fermentation. The primary precursor for EC formation is urea, which originates from both external sources and arginine degradation. Urea degradation is suppressed by nitrogen catabolite repression (NCR) in Saccharomyces cerevisiae. The regulation of NCR is mediated by two positive regulators (Gln3p, Gat1p/Nil1p) and two negative regulators (Dal80p/Uga43p, Deh1p/Nil2p/GZF3p). DAL80 revealed higher transcriptional level when yeast cells were cultivated under nitrogen-limited conditions. In this study, when DAL80-deleted yeast cells were compared to wild-type BY4741 cells, less urea was accumulated, and genes involved in urea utilization were up-regulated. Furthermore, Chinese rice wine fermentation was conducted using dal80Δ cells; the concentrations of urea and EC were both reduced when compared to the BY4741 and traditional fermentation starter. The findings of this work indicated Dal80p is involved in EC formation possibly through regulating urea metabolism and may be used as the potential target for EC reduction. Full article

The aim of this work was to clarify the formation of ethyl carbamate (EC) and its influence factors throughout the production process of Cantonese soy sauce. The results showed that EC was not detected in the koji-making and early moromi fermentation stages, but started to be generated when pH of the moromi decreased to about 4.9—at the same time, the levels of ethanol, urea and citrulline increased significantly. Most EC was formed during raw soy sauce hot extraction (40.6%) and sterilization (42.9%) stages. The EC content exhibited the highest correlation with ethanol throughout the whole production process (R = 0.97). The simulation soy sauce produced in laboratory led the same conclusion—moreover, the contents of EC, ethanol and citrulline were higher in soy sauce fermented at 30 °C than in soy sauce fermented at 15 °C. Extraction of raw soy sauce by squeezing contributed little to EC formation. Further research showed that citrulline and ethanol led to significant increases in EC levels in raw soy sauce upon heating. These results indicate that ethanol and citrulline are two critical precursors of EC and that EC is mainly formed during the heat treatment stage of soy sauce. Full article

The most common way to produce red wine is through the use of Saccharomyces cerevisiae strains for alcoholic fermentation and lactic acid bacteria for malolactic fermentation. This traditional winemaking methodology produces microbiologically stable red wines. However, under specific conditions off-flavours can occur, wine quality can suffer and human health problems are possible, especially after the second fermentation by the lactic acid bacteria. In warm countries, problems during the malolactic fermentation arise because of the high pH of the must, which makes it very difficult to properly control the process. Under such conditions, wines with high acetic acid and histamine concentrations are commonly produced. This study investigates a recent red
wine-making technology that uses a combination of Lachancea thermotolerans and Schizosaccharomyces pombe as an alternative to the conventional malolactic fermentation. This work studies new parameters such as aroma compounds, amino acids, ethanol index and sensory evaluation. Schizosaccharomyces pombe totally consumes malic acid while Lachancea thermotolerans produces lactic acid, avoiding excessive deacidification of musts with low acidity in warm viticulture areas. This methodology also reduces the malolactic fermentation hazards in wines with low acidity. The main products are wines that contain less acetic acid, less biogenic amines and precursors and less ethyl carbamate precursors than the traditional wines produced via conventional fermentation techniques. Full article

The lithiation of 2-(2,2-dimethyl-1,3-dioxolan-4-yl)ethyl diisopropylcarbamate (1) is achieved freely by sec-butyllithium in diethylether with high lk-diastereoselectivity: the bicyclic chelate complexes 3a and 3b are reacted with electrophiles to form optically active precursors 4a and 4b with >95% diastereoselectivity. In addition, tertiary diamines can undergo an external complexation in contest with the internal oxygen ligand, leading to improved stereoselectivities. The further reactions of lithiated carbamates with trans alkenyl-9-BBN derivatives after 1,2 metallate rearrangements, gave the key intermediate α-substituted allylic boranes 7. Subsequent allylboration of aldehydes gave (Z)-anti-homoallylic alcohols 8 in good yield and excellent d.r. Full article