The synthetic community of lactic acid bacteria (LAB) is commonly utilized in the food industry for manipulating product properties. However, the intermediate interactions and ecological stability resulting from metabolic differences among various LAB types remain poorly understood. We aimed to analyze the metabolic behavior of single and combined lactic acid bacteria in China rice wine based on microbial succession. Three-stage succession patterns with obligate heterofermentative LAB dominating prefermentation and homofermentative LAB prevailing in main fermentation were observed. Facultative heterofermentative LAB exhibited significant growth. Pairwise coculture interactions revealed 63.5% positive, 34.4% negative, and 2.1% neutral interactions, forming nontransitive and transitive competition modes. Nontransitive competitive combinations demonstrated stability over ∼200 generations through amino acid (mainly aspartic acid, glutamine, and serine) cross-feeding and lactic acid detoxification, which also showed potential for controlling biogenic amines and developing LAB starter cultures. Our findings offer insights into the mechanistic underpinnings of LAB interaction networks.

Urethane hydrolase can degrade the carcinogen ethyl carbamate (EC) in fermented food, but its stability and activity limit its application. In this study, a mutant G246A and a double mutant N194V/G246A with improved cpUH activity and stability of Candida parapsilosis were obtained by site-directed mutagenesis. The catalytic efficiency (Kcat/Km) of mutant G246A and double mutant N194V/G246A are 1.95 times and 1.88 times higher than that of WT, respectively. In addition, compared with WT, the thermal stability and pH stability of mutant G246A and double mutant N194V/G246A were enhanced. The ability of mutant G246A and double mutant N194V/G246A to degrade EC in rice wine was also stronger than that of WT. The mutation increased the stability of the enzyme, as evidenced by decreased root mean square deviation (RMSD) and increased hydrogen bonds between the enzyme and substrate by molecular dynamics simulation and molecular docking analysis. The molecule modification of new cpUH promotes the industrial process of EC degradation.

This research paper focuses on the application of the \"Design-Build-Test-Learn\" framework to design and evaluate a synthetic microbial community aimed at studying the impact of Lactic Acid Bacteria (LAB) interactions and fitness on the formation of biogenic amines (BAs) in Chinese rice wine (CRW). The study reveals a close correlation between the assembly model of LAB and the accumulation of BAs in CRW, and multiple interactions were observed between amine-producing and non-amine-producing LAB, including commensalism, amensalism, and competition. The commensalism among amine-producing LAB was found to promote BAs accumulation through metabolic cross-feeding of amino acids. Moreover, the higher-order interaction community was designed to regulate the BAs formation effectively. For instance, the interference of Lactiplantibacillus plantarum (ACBC271) resulted in the elimination of amine-producing LAB viability, resulting in a 22% decrease (not exceeding 43.54 mg/L) in the total amount of BAs. Simulation of community dynamics models further suggests that LAB with quantitative social interactions can effectively control LAB accumulation in CRW by forecasting fluctuation in BAs generation through fitness competition and metabolic interference. Overall, this study provides valuable insights into the complex interaction networks within microbial communities in traditional fermentation ecosystems. It also proposes a novel approach for quality control of nitrogen food safety factors in fermented foods.

BACKGROUND: It has been well accepted that lactic acid bacteria (LAB) are the main bacterial genera present during the brewing of Chinese rice wine (CRW). LAB plays a decisive role in the flavor quality of CRW; however, its application in CRW has previously been overlooked. Therefore, effects of different LAB as co-fermenter on the flavor characteristics of CRW were investigated.
RESULTS: Co-fermentation of LAB increased the utilization rate of reducing sugar, concentration of lactic acid, amino acid nitrogen and total acidity, as well as the content of volatile flavor compounds. Different LAB doses had little effect on the flavor profiles of CRW, but the species of LAB greatly affected the flavor characteristic. The flavor of CRW co-fermented with Lactococcus lactis was characterized by long-chain fatty acid ethyl esters, while co-fermentation with Weissella confusa highlighted the ethyl esters of low molecular weight and short carbon chains in the resultant CRW. Alcohol compounds were dominant in the CRW co-fermented using Pediococcus pentosaceus.
CONCLUSIONS: The co-fermentation of LAB increased the number of volatile flavor compounds, especially esters. LAB exhibited great potential in the application of CRW industry to enrich the flavor characteristics and enhance the flavor diversity of the final product. © 2023 Society of Chemical Industry.

Aging is an essential operation to perfect the flavor quality of Hungjiu. In this study, formation mechanism of flavor compounds responsible for the characteristic flavor of aged Huangjiu was investigated. The contents of umami and bitter free amino acids (FAA) increased with the storage period prolonged, while that of sweet FAA showed downward trend. Gas chromatograph-mass spectrometry and principal component analysis indicated that the volatile flavor compounds with OAV exceed 1, especially middle-chain fatty-acid-ethyl-esters and aromatic compounds, dominated the characteristic flavor of aged Huangjiu. Low field-NMR was firstly applied to characterize the molecular association between water and dissolved flavor compounds in aged Huangjiu. The results showed that basic amino acids contributed greatly to the flavor formation of aged Huangjiu via molecular association. In addition, the molecular association significantly promoted the accumulation of flavor compounds with OAV > 1, especially ethyl esters.

BACKGROUND: Ethyl carbamate (EC) is a potential carcinogen existing in fermented foods such as Chinese rice wine (Huangjiu). Since urea is an important precursor of EC, the degradation of urea could be an effective way to reduce EC in foods.
RESULTS: In this study, an Enterobacter sp. R-SYB082 with acid urea degradation characteristics was obtained through microbial screening. Further research isolated a new acid urea-degrading enzyme from R-SYB082 strain - ureidoglycolate amidohydrolase (UAH) - which could degrade EC directly. The cloning and expression of UAH in Escherichia coli BL21 (DE3) suggested that the activity of urea-degrading enzyme reached 3560 U L-1 , while urethanase activity reached 2883 U L-1 in the optimal fermentation condition. The enzyme had the dual ability of degrading substrate urea and product EC. The removal rate of EC in Chinese rice wine could reach 90.7%.
CONCLUSIONS: This study provided a new method for the integrated control of EC in Chinese rice wine and other fermented foods. © 2022 Society of Chemical Industry.

In order to improve the high cost of equipment and difficult management caused by the natural aging of Chinese rice wine (Huangjiu), micro-oxygen (MO) and electric field (PEF) technology are used to accelerate the aging of Huangjiu. The results showed that micro-oxygen and electric field have a significant effect on the sensory characteristics and flavor characteristics of Huangjiu. Compared with the naturally aged Huangjiu, the flavor compounds of Huangjiu treated with micro-oxygen and electric field increase significantly. Based on principal component analysis, Huangjiu processed at 0.35 mg L/day or 0.5 mg L/day combined electric field exhibited similar flavor to the natural aged Huangjiu, which was highly associated with long-chain fatty acid ethyl esters (C13-C18). Moreover, partial least squares regression demonstrated that sensory attributes of cereal aroma and astringency were highlighted after aging time, while fruit aroma, continuation, and full body were dominant after micro-oxygen and electric field treatment. Micro-oxygen and electric field effectively enhanced the quality of Huangjiu, which could be applied in other alcoholic beverages.

In this study, an artificial starter culture was prepared using the core microbial species of JIUYAO to produce Chinese rice wine (CRW). The fermentation activity and flavor profiles of CRW samples fermented with traditional JIUYAO, a commercial starter culture, and our artificial starter culture were compared. The optimal protectant combination for lyophilization of the artificial starter was established as 15.09% skim milk, 4.45% polyethylene glycol, 1.96% sodium glutamate, and 11.81% maltodextrin. A comparative analysis revealed that the ethanol content of the three CRW samples was similar. The total acid content of the CRW sample fermented with the artificial starter (7.10 g/L) was close to that of the sample fermented with JIUYAO (7.35 g/L), but higher than that of the sample fermented with the commercial starter (5.40 g/L). An electronic nose analysis revealed that the olfactory fingerprints of the CRW samples fermented with JIUYAO and the artificial starter resembled each other. For both above mentioned samples, the flavor profiles determined by gas chromatography-mass spectrometry indicated some differences in the variety and content of the aroma compounds, but the key odorants (odor activity values ≥1), such as isoamyl acetate, ethyl acetate, phenyl alcohol, and isoamyl alcohol, were similar.

The traditional low-alcoholic beverages, such as grape wine, sake, and rice wine, have been consumed all over the world for thousands of years, each with their unique methods of production that have been practiced for centuries. Moderate consumption of wine is generally touted as beneficial for health, although there is ongoing debate for the responsible components in wine. In this review, the structural and functional characteristics, the formation mechanisms, and their health-promoting activities of peptides in three brewed wines, grape wine, Chinese rice wine (also called Chinese Huangjiu or Chinese yellow wine), and Japanese sake, are discussed. The formation of peptides in wine imparts sensorial, technological, and biological attributes. Prospects on future research, with an emphasis on the peptide characterization, formation mechanism, physiological activity, and molecular mechanisms of action, are presented.

The study quantitatively profiled 83 low-molecular-weight metabolites in the categories of alcohols, aldehydes, amino acids, esters, fatty acids, organic acids, and reducing sugars produced during the advanced brewing process of Chinese rice wine, using multiply chromatography and mass spectrum. In the primary fermentation, vigorous metabolisms were demonstrated by the production of ethanol at the level of 14% by volume, and the consumption of reducing sugars from the maximum level of 100 g/L to 20 g/L. Meantime, more than 70% of the contents of organic acids, fatty acids, higher alcohols, and aldehydes were formed. The metabolisms slowed down in the secondary fermentation, whereas 60% of the contents of amino acids and esters were accumulated in this stage. The nutrients, such as amino acids, organic acids, and reducing sugars reached 10 g/L, 5 g/L, and 3 g/L at the end of brewing, respectively. In terms of flavor and taste attributes to the brewed rice wine, the organoleptic activity value (i.e. the ratio of content to threshold value) was above 1 for 17 compounds, including six organic acids, namely acetic acid, citric acid, lactic acid, malic acid, succinic acid, and tartaric acid, four amino acids, namely cysteine, aspartic acid, glutamic acid, and lysine, three higher alcohols namely isoamyl alcohol, isobutanol, and phenethyl alcohol, three esters, namely ethyl acetate, ethyl butyrate, and ethyl hexanoate, and an aldehyde, namely benzaldehyde.