Effect of a co-fermentation system with high-GABA-yielding strains on soymilk properties: microbiological, physicochemical, and aromatic characterisations.

Biosynthesis is the safest method for preparing GABA; however, there are not enough GABA-producing strains to provide an effective resource. The purpose of this study was to determine the feasibility of using Lactobacillus fermentum SMN10-3(A) and Lactococcus lactis SMN15-6(B) to study the effects of strain complex pairing on the GABA formation, flavour, and metabolic pathways of fermented soymilk. It was found that group A2B1 had the highest acid production rate, GABA yield (1.76 ± 0.01 mg/mL), and flavour compound content. A total of 55 differential metabolites were produced after fermentation, of which 28 dominated by hexanal were significantly downregulated and 26 dominated by alcohols were significantly upregulated. The significant metabolic pathways involved were d-alanine, taurine and hypotaurine, and selenocompound metabolism. Finally, the components contributing to the aroma of fermented soymilk were identified, which included 2-pentylfuran and 2-butyl-2-octenal. These results provide a theoretical basis for future research on GABA-rich fermented foods.

LABAMPsGCN: A framework for identifying lactic acid bacteria antimicrobial peptides based on graph convolutional neural network.

Lactic acid bacteria antimicrobial peptides (LABAMPs) are a class of active polypeptide produced during the metabolic process of lactic acid bacteria, which can inhibit or kill pathogenic bacteria or spoilage bacteria in food. LABAMPs have broad application in important practical fields closely related to human beings, such as food production, efficient agricultural planting, and so on. However, screening for antimicrobial peptides by biological experiment researchers is time-consuming and laborious. Therefore, it is urgent to develop a model to predict LABAMPs. In this work, we design a graph convolutional neural network framework for identifying of LABAMPs. We build heterogeneous graph based on amino acids, tripeptide and their relationships and learn weights of a graph convolutional network (GCN). Our GCN iteratively completes the learning of embedded words and sequence weights in the graph under the supervision of inputting sequence labels. We applied 10-fold cross-validation experiment to two training datasets and acquired accuracy of 0.9163 and 0.9379 respectively. They are higher that of other machine learning and GNN algorithms. In an independent test dataset, accuracy of two datasets is 0.9130 and 0.9291, which are 1.08% and 1.57% higher than the best methods of other online webservers.

Novel Insight into the Formation of Odour-Active Compounds in Sea Buckthorn Wine and Distilled Liquor Based on GC-MS and E-Nose Analysis.

Sea buckthorn wine (SW) and distilled liquor (DL) are fruit wines with beneficial health effects. However, their unpleasant flavour limits their development and widespread acceptance. Therefore, it is necessary to analyse their flavour composition and changes. In this study, differential metabolites of sea buckthorn DL during processing were analysed, and the relationships between E-nose sensor values and key volatile organic compounds (VOCs) were established. The results show that 133 VOCs were identified, with 22 aroma-contributing components. Fermentation significantly increased the content of VOCs, especially esters. A total of seven and 51 VOCs were significantly upregulated after fermentation and distillation, respectively. Meanwhile, seven sensors were positively correlated with the increased level of alcohols and esters, and reflected the increasing trends of 10 key VOCs.

Microbial diversity analysis of jiaoke from Xilingol, Inner Mongolia.

Jiaoke is a traditional Mongolian fermented dairy product that is nutritious and has a unique taste. It is made from the fat separated from fermented milk. In this study, we collected 24 jiaoke samples from the Xilingol region of Inner Mongolia. The microbiota composition of the collected samples was analyzed using 16S rRNA small-molecule real-time sequencing, and the lactic acid bacteria (LAB) population was enumerated and isolated by laboratory culture techniques. We used an electronic tongue device to assess the taste quality of the products. One hundred fifty LAB isolates (5 genera and 14 species) were recovered and identified by 16S rRNA sequencing across all samples. Lactococcus lactis and Lactobacillus plantarum accounted for 51.33% and 10.67% of the total isolates, respectively. The small-molecule real-time sequencing of full-length 16S rRNAs revealed an overall bacterial microbiota composition of 10 phyla, 121 genera, and 186 species, largely represented by sequences of Lactococcus (68.46%) and Lactococcus lactis (52.92%) at the genus and species levels, respectively. The electronic tongue analysis revealed that the sweetness, bitterness, sourness, and saltiness of jiaoke varied greatly between samples. The presence of Lactococcus lactis correlated positively with bitter aftertaste; the presence of Lactococcus piscium correlated positively with umami and negatively with astringent and bitter aftertastes; and the presence of Lactobacillus helveticus correlated positively with sourness and negatively with other taste qualities. These results suggest that the microbiota composition and product taste are closely related. The novel LAB strains collected in this work represent valuable natural microbial resources.

A survey of the relationship between functional genes and acetaldehyde production characteristics in Streptococcus thermophilus by multilocus sequence typing.

Streptococcus thermophilus is an important bacterium used in the production of fermented dairy products. Yogurt with good flavor is preferred by consumers; thus, variation in flavor-formation characteristics among isolates is attracting attention. Here, acetaldehyde production characteristics of 30 isolates were evaluated in parallel with genotyping and multilocus sequence typing of key functional genes involved in acetaldehyde production. The results showed that isolates could be divided into 3 phenotypically distinct groups: high-acetaldehyde-yielding isolates (>10 mg/L), medium-acetaldehyde-yielding isolates (5-10 mg/L) and low-acetaldehyde-yielding (<5 mg/L) based on evaluation of acetaldehyde production during yogurt storage. These groups, distinguishable by phenotypic characteristics, were clustered in corresponding groups based on functional gene multilocus sequence typing analysis. Combining functional gene sequence analysis of 30 Strep. thermophilus isolates with phenotypic evaluation of their flavor-related characteristics (specifically acetaldehyde production) demonstrated that groups of isolates established using genotype data analysis corresponded with groups identified based on their phenotypic traits. Interestingly, the 30 isolates of Strep. thermophilus showed significant phylogenetic clustering in acetaldehyde content by functional gene and acetaldehyde content analysis. A corresponding relationship exists between functional gene phylogenetic clustering and acetaldehyde content variation.

Novel Method for l-Methionine Production Catalyzed by the Aminotransferase ARO8 from Saccharomyces cerevisiae.

The aminotransferase ARO8 was proved to play an efficient role in conversion of l-methionine into methionol via the Ehrlich pathway in Saccharomyces cerevisiae in our previous work. In this work, the reversible transamination activity of ARO8 for conversion of α-keto-γ-(methylthio) butyric acid (KMBA) into l-methionine was confirmed in vitro. ARO8 was cloned from S. cerevisiae S288c and overexpressed in Escherichia coli BL21. A 2-fold higher aminotransferase activity was detected in the recombinant strain ARO8-BL21, and ARO8 was detected in the supernatant of ARO8-BL21 lysate with IPTG induction by SDS-PAGE analysis. The recombinant ARO8 was then purified and used for transforming KMBA into l-methionine. An approximately 100% conversion rate of KMBA into l-methionine was achieved by optimized enzymatic reaction catalyzed by ARO8. This work fulfilled l-methionine biosynthesis catalyzed by the aminotransferase ARO8 using glutamate and KMBA, which provided a novel method for l-methionine production by enzymatic catalysis with the potential application prospect in industry.

Improving flavor metabolism of Saccharomyces cerevisiae by mixed culture with Wickerhamomyces anomalus for Chinese Baijiu making.

Yeasts are the most important microorganisms in the fermentation of Chinese Baijiu and the interaction of these yeasts could impact the quality of Baijiu. In this study, we initially characterized the Baijiu yeasts and evaluated their fermentation potential. Wickerhamomyces anomalus GZ3 and Saccharomyces cerevisiae G20 were found to generate high yields of ethyl acetate (2.76 g/L) and 2-phenylethanol, respectively. Results also indicated that the use of W. anomalus along with S. cerevisiae increased volatile compounds production, the maximum ethyl acetate production was observed in S. cerevisiae and W. anomalus at 106:106 ratio and have increased by 33 % compared with single culture of W. anomalus. Besides, there was a significant increase of 2-phenylethanol (3.29 g/L) production in single culture of S. cerevisiae with the addition of l-phenylalanine. However, the conversion of l-phenylalanine in mixed culture had significant impact on the yeasts interaction and end flavor of Baijiu. Thus, the present study provided new insights into yeasts interactions in Baijiu fermentation and the effect of some primary metabolites on the end flavor and Baijiu quality.

Generation of 2-Furfurylthiol by Carbon-Sulfur Lyase from the Baijiu Yeast Saccharomyces cerevisiae G20.

2-Furfurylthiol is the representative aroma compound of Chinese sesame-flavored baijiu. Previous studies demonstrated that baijiu yeasts could generate 2-furfurylthiol using furfural and l-cysteine as precursors and that the Saccharomyces cerevisiae genes STR3 and CYS3 are closely related to 2-furfurylthiol biosynthesis. To confirm the mechanism of the STR3- and CYS3-gene products on 2-furfurylthiol biosynthesis, their encoded proteins were purified, and we confirmed their activities as carbon-sulfur lyases. Str3p and Cys3p were able to cleave the cysteine-furfural conjugate to release 2-furfurylthiol. Moreover, the characterization of the enzymatic properties of the purified proteins shows good thermal stabilities and wide pH tolerances, which enable their strong potential for various applications. These data provide direct evidence that yeast Str3p and Cys3p release 2-furfurylthiol in vitro, which can be applied to improve baijiu flavor.

STR3 and CYS3 Contribute to 2-Furfurylthiol Biosynthesis in Chinese Sesame-Flavored Baijiu Yeast.

2-Furfurylthiol is an important aroma compound with characteristic sesame flavor. It was recently identified as the representative aroma compound of Chinese sesame-flavored Baijiu. But its formation mechanism is unclear. In this study, the dominant yeast species Saccharomyces cerevisiae, Pichia kudriavzevii, and Wickerhamomyces anomalus were isolated from samples of Chinese sesame-flavored fermentation and were screened for the capacity of 2-furfurylthiol biosynthesis. Some isolates were found to be capable of generating 2-furfurylthiol using furfural and l-cysteine as precursors, among which S. cerevisiae G20 exhibited the strongest capacity with a yield of 3.03 mg/L. Furthermore, the genes STR3 and CYS3 from S. cerevisiae were cloned and overexpressed in the reference strain S. cerevisiae S288c and the isolate strain G20, respectively, which resulted in a significant increase in 2-furfurylthiol release in the two strains. Moreover, inactivation of STR3 gene in S288c and G20 led to obvious reduction in 2-furfurylthiol production, demonstrating that STR3 got involved in 2-furfurylthiol biosynthesis in S. cerevisiae. It is the first time that the yeast isolates with high capacity of 2-furfurylthiol biosynthesis were found during Chinese sesame-flavored Baijiu fermentation and confirmed that the genes STR3 and CYS3 were closely relevant to 2-furfurylthiol biosynthesis.

Relationships between functional genes in Lactobacillus delbrueckii ssp. bulgaricus isolates and phenotypic characteristics associated with fermentation time and flavor production in yogurt elucidated using multilocus sequence typing.

Lactobacillus delbrueckii ssp. bulgaricus (L. bulgaricus) is well known for its worldwide application in yogurt production. Flavor production and acid producing are considered as the most important characteristics for starter culture screening. To our knowledge this is the first study applying functional gene sequence multilocus sequence typing technology to predict the fermentation and flavor-producing characteristics of yogurt-producing bacteria. In the present study, phenotypic characteristics of 35 L. bulgaricus strains were quantified during the fermentation of milk to yogurt and during its subsequent storage; these included fermentation time, acidification rate, pH, titratable acidity, and flavor characteristics (acetaldehyde concentration). Furthermore, multilocus sequence typing analysis of 7 functional genes associated with fermentation time, acid production, and flavor formation was done to elucidate the phylogeny and genetic evolution of the same L. bulgaricus isolates. The results showed that strains significantly differed in fermentation time, acidification rate, and acetaldehyde production. Combining functional gene sequence analysis with phenotypic characteristics demonstrated that groups of strains established using genotype data were consistent with groups identified based on their phenotypic traits. This study has established an efficient and rapid molecular genotyping method to identify strains with good fermentation traits; this has the potential to replace time-consuming conventional methods based on direct measurement of phenotypic traits.

Study on Streptococcus thermophilus isolated from Qula and associated characteristic of acetaldehyde and diacetyl in their fermented milk.

In this study, the lactic acid bacterial population of Qula cheese from the Gansu and Sichuan provinces of China were isolated and identified. Eight strains of Streptococcus thermophilus were isolated, of which five strains were selected for further characterization based on their fermentation properties. The changes in a number of parameters, including titration acidity, pH, viable counts, PrtS protease activity and the production of acetaldehyde, diacetyl and organic acid, were monitored during fermentation and the storage of fermented milks produced by the respective strain. All of the strains displaying acidifying capacity and all five fermented milks maintained high viable counts of S. thermophilus from fermentation to storage. Our study found that the changes in the monitored parameters were strain-specific and varied considerably among the five tested strains. Fermented milks produced by strain IMAU80809 had the highest concentration of acetaldehyde and were most favorable in the sensory evaluation. This study confirms that Qula cheese is a good source for isolating novel lactic acid bacterial strains with different fermentation properties, which will be very useful for further development and industrialization of traditionally fermented dairy products.