Ultrasound-assisted fermentation for antioxidant peptides preparation from okara: Optimization, stability, and functional analyses.

This study investigated the effects of ultrasound-assisted fermentation (UAF) on the preparation of antioxidant peptides (UAFP) from okara and examined their content, chemical structures, and antioxidant activity. After the optimal ultrasonic processing (time, 20 min; frequency, 45 KHz; power, 120 W/L), the peptide content yield reached the maximum of 12.36 ± 0.02 mg/mL, and their DPPH free radical scavenging rate was 65.15 ± 0.32 %. UAF increased the number of globular aggregates with deeper gullies, a looser structure, and higher porosity. The experiments conducted using the oxidative stress injury model of HepG2 cells showed that okara UAFP promoted cell growth and exerted a protective effect. Moreover, ultrasonic treatment remarkably improved the environmental stability (NaCl, glucose, sodium benzoate, temperature, pH, metal ions) and antioxidant activity of UAFP. Concisely, optimal ultrasonic processing can aid the fermentation of agroindustrial by-products to prepare antioxidant peptides, such as natural food antioxidant peptides from soybean waste.

Antimicrobial peptides: An alternative to traditional antibiotics.

As antibiotic-resistant bacteria and genes continue to emerge, the identification of effective alternatives to traditional antibiotics has become a pressing issue. Antimicrobial peptides are favored for their safety, low residue, and low resistance properties, and their unique antimicrobial mechanisms show significant potential in combating antibiotic resistance. However, the high production cost and weak activity of antimicrobial peptides limit their application. Moreover, traditional laboratory methods for identifying and designing new antimicrobial peptides are time-consuming and labor-intensive, hindering their development. Currently, novel technologies, such as artificial intelligence (AI) are being employed to develop and design new antimicrobial peptide resources, offering new opportunities for the advancement of antimicrobial peptides. This article summarizes the basic characteristics and antimicrobial mechanisms of antimicrobial peptides, as well as their advantages and limitations, and explores the application of AI in antimicrobial peptides prediction amd design. This highlights the crucial role of AI in enhancing the efficiency of antimicrobial peptide research and provides a reference for antimicrobial drug development.

Identification, Characterization, and Receptor Binding Mechanism of New Umami Peptides from Traditional Fermented Soybean Paste (Dajiang).

Dajiang, a traditional Chinese condiment, is made from fermented soybeans. It is highly popular among consumers as a result of its delicious umami flavor, which mainly originates from umami peptides. To examine the mechanism of umami taste in Dajiang, we selected Dajiang samples with strong umami taste and subjected them to purification and identification analysis using ethanol precipitation, gel chromatography, reversed-phase high-performance liquid chromatography, and ultraperformance liquid chromatography-tandem mass spectrometry. Subsequently, on the basis of toxicity and umami prediction analysis, we screened, synthesized, and characterized three novel bean umami peptides in Dajiang: TLGGPTTL, 758.4174 Da; GALEQILQ, 870.4811 Da; and HSISDLQ, 911.4713 Da. Their sensory threshold values were 0.25, 0.40, and 0.17 mmol/L, respectively. Furthermore, molecular docking results showed that hydrogen-bonding and hydrophobic interactions are important interaction forces in the binding of umami peptide to taste receptors. Ser147 and Glu148 of the T1R3 taste receptor are important amino acid residues for binding of the three umami peptides. This study uncovers the mechanism of umami-peptide-driven flavor in fermented soybean products.

A systematic review on the flavor of soy-based fermented foods: Core fermentation microbiome, multisensory flavor substances, key enzymes, and metabolic pathways.

The characteristic flavor of fermented foods has an important impact on the purchasing decisions of consumers, and its production mechanisms are a concern for scientists worldwide. The perception of food flavor is a complex process involving olfaction, taste, vision, and oral touch, with various senses contributing to specific properties of the flavor. Soy-based fermented products are popular because of their unique flavors, especially in Asian countries, where they occupy an important place in the dietary structure. Microorganisms, known as the souls of fermented foods, can influence the sensory properties of soy-based fermented foods through various metabolic pathways, and are closely related to the formation of multisensory properties. Therefore, this review systematically summarizes the core microbiome and its interactions that play an active role in representative soy-based fermented foods, such as fermented soymilk, soy sauce, soybean paste, sufu, and douchi. The mechanism of action of the core microbial community on multisensory flavor quality is revealed here. Revealing the fermentation core microbiome and related enzymes provides important guidance for the development of flavor-enhancement strategies and related genetically engineered bacteria.

Recent developments in horizontal gene transfer with the adaptive innovation of fermented foods.

Horizontal gene transfer (HGT) has contributed significantly to the adaptability of bacteria, yeast and mold in fermented foods, whose evidence has been found in several fermented foods. Although not every HGT has biological significance, it plays an important role in improving the quality of fermented foods. In this review, how HGT facilitated microbial domestication and adaptive evolution in fermented foods was discussed. HGT can assist in the industrial innovation of fermented foods, and this adaptive evolution strategy can improve the quality of fermented foods. Additionally, the mechanism underlying HGT in fermented foods were analyzed. Furthermore, the critical bottlenecks involved in optimizing HGT during the production of fermented foods and strategies for optimizing HGT were proposed. Finally, the prospect of HGT for promoting the industrial innovation of fermented foods was highlighted. The comprehensive report on HGT in fermented foods provides a new trend for domesticating preferable starters for food fermentation, thus optimizing the quality and improving the industrial production of fermented foods.

Identification, taste characterization, and molecular docking study of a novel microbiota-derived umami peptide.

Umami peptides are an important taste substance in fermented foods. However, in the absence of known microbiota-derived umami peptides, the understanding of the umami mechanism remains unclear. Tetragenococcus halophilus, a dominant fermentation bacteria, may be an important source of umami peptides. Accordingly, T. halophilus fermentation broth was fractioned by ethanol precipitation, gel chromatography, and reverse phase-high performance liquid chromatography. The isolated peptide fraction with the most intense umami taste was screened by amino acid composition and sensory analyses. Finally, three novel microbiota-derived peptides (DFE, LAGE, and QLQ) were identified, synthesized, and characterized for taste. Among them, only DFE had umami and umami-enhancing abilities improving multiple tastes. Molecular docking studies indicated that DEF binds to T1R1/T1R3 receptors through hydrogen bonding and electrostatic interactions involving receptor residues Ser332, Ser256, ASN41, His125, etc. This study highlights the critical role of microbiota-derived peptides in the umami taste of fermented foods.

Extraction, purification, structural characterization, and antioxidant activity of a novel polysaccharide from Lonicera japonica Thunb.

A novel water-soluble polysaccharide (HEP-4) with a molecular weight of 1.98 × 105Da was extracted from honeysuckle. Structural characterization was performed using high-performance liquid chromatography (HPLC), gas chromatography, Fourier transform-infrared (FT-IR) spectrum, nucleus magnetic resonance (NMR) spectra, and scanning electron microscopy. The results showed that HEP-4 is primarily composed of mannose, rhamnose, galacturonic acid, glucose, galactose, and arabinose with a mole ratio of 6.74:1.56:1.04:14.21:4.31:5.4, and the major types of the glycosidic bond types of HEP-4 were 1-α-D-Glcp, 1,4-ß-D-Glcp, 1-ß-D-Arap, 1,3,4-ß-D-Arap, and 1,3,6-ß-D-Manp. The results of bioactivity experiments revealed that HEP-4 had antioxidant in vitro. In addition, HEP-4 inhibited H2O2-induced oxidative damage and increased the activity of HepG2 cells by reducing MDA levels and inhibiting ROS production. Meanwhile, HEP-4 significantly enhanced the activities of GSH-Px and CAT, indicating that HEP-4 exerts a protective effect on H2O2-induced oxidative stress. These results indicate that HEP-4 could be a potential natural antioxidant.

Structure characterization, antioxidant and emulsifying capacities of exopolysaccharide derived from Tetragenococcus halophilus SNTH-8.

Tetragenococcus halophilus exopolysaccharides (THPS) are metabolites released by T. halophilus SNTH-8 to resist a high-salt environment. Although many studies have investigated the mechanisms underlying salt tolerance shown by T. halophilus, structural characteristics as well as antioxidant and emulsifying capacities of THPS remain unclear. In this study, we isolated and purified two components, THPS-1 and THPS-2, from T. halophilus SNTH-8. Purified THPS-1 and THPS-2 were composed of arabinose, xylose, fucose, galactose, glucose, and glucuronic acid at a molar ratio of 1.66:38.95:2.11:26.12:29.73:1.43 and 0.46:40.3:0.54:30.8:1.36:25.54, respectively. The average molecular weights of THPS-1 and THPS-2 were 14.98 kDa and 21.03 kDa, respectively. Moreover, the structures of THPS-1 and THPS-2 were investigated via fourier-transform infrared spectroscopy(FT-IR), nuclear magnetic resonance spectroscopy(NMR), scanning electron microscopy(SEM), and methylation analysis. THPS-1 was a highly branched polysaccharide with a backbone of α-D-(1,4)-Xyl, α-D-(1,6)-Glc and α-D-Xyl as the terminal, while THPS-2 was a highly branched polysaccharide with a backbone of α-D-(1,4)-Xyl and ß-D-GlcA as the terminal. The branches were identified as ß-D-(1,4,6)-Gal and ß-D-(1,6)-Gal. Both THPS-1 and THPS-2 exhibited high antioxidant and emulsifying capacities. Overall, our structural analysis of THPS may further enhance research on natural emulsifiers and antioxidants.

Advances in fermented foods revealed by multi-omics: A new direction toward precisely clarifying the roles of microorganisms.

Fermented foods generally comprise a complex micro-ecosystem with beneficial microbiota, functional products, and special flavors and qualities that are welcomed globally. Single-omics analysis allows for a comprehensive characterization of the main microbial factors influencing the function, flavor, and quality of fermented foods. However, the species, relative abundance, viability, growth patterns, and metabolic processes of microorganisms vary with changes in processing and environmental conditions during fermentation. Furthermore, the mechanisms underlying the complex interaction among microorganisms are still difficult to completely understand and analyze. Recently, multi-omics analysis and the integration of multiple types of omics data allowed researchers to more comprehensively explore microbial communities and understand the precise relationship between fermented foods and their functions, flavors, and qualities. Multi-omics approaches might help clarify the mechanisms underpinning the fermentation processes, metabolites, and functional components of these communities. This review clarified the recent advances in the roles of microorganisms in fermented foods based on multi-omics data. Current research achievements may allow for the precise control of the whole industrial processing technology of fermented foods, meeting consumers' expectations of healthy products.

Investigating the core microbiota and its influencing factors in traditional Chinese pickles.

Pickles are a type of traditional fermented food in Northeast China that exhibit a broad variety of preparations, flavors and microbial components. Despite their widespread consumption, the core microorganisms in various traditional pickles and the precise impact of ecological variables on the microbiota remains obscure. The present study aims to unravel the microbial diversity in different pickle types collected from household (12 samples) and industrial (10 samples) sources. Among these 22 samples tested, differences were observed in total acid, amino acid nitrogen, nitrite, and salt content. Firmicutes and Ascomycota emerged as the predominant microbial phyla as observed by Illumina MiSeq sequencing. Amongst these, the commonly encountered microorganisms were Lactobacillus, Weissella and yeast. Comparative analysis based on non-metric multidimensional scaling (NMDS), showed that the microbial community in the pickles was affected by external conditions such as major ingredients and manufacturing process. Correlation analysis further showed that the resident core microorganisms in pickles could adapt to the changing internal fermentation environment. The insights gained from this study further our understanding of traditional fermented foods and can be used to guide the isolation of excellent fermented strains.

Metatranscriptome-based investigation of flavor-producing core microbiota in different fermentation stages of dajiang, a traditional fermented soybean paste of Northeast China.

Dajiang, or naturally fermented soybean paste, has a unique flavor that is influenced by the resident microflora. However, the association between flavor and the core microbiota is unclear. Recent advances in RNA sequencing have identified genes that are actively expressed in complex microbial communities. To this end, we analyzed the time-dependent changes in the microbiota and the metabolite profiles of Dajiang using metatranscriptome sequencing, HS-SPME-GC-MS and amino acid analysis identified 10 volatile compounds that contribute to the development of soybean paste flavor. Further analysis of the correlation between the active microorganisms and the physicochemical characteristics and flavor substances in soybean paste indicated that Lactobacillus and Tetragenococcus were the core genera affecting chromaticity and flavor. These microorganisms produce enzymes that catalyze a series of metabolic pathways that generate flavor substances. Our findings provide new insights into the role of the microbiota in the development of flavor in fermented foods.

Determination of allosteric and active sites responsible for catalytic activity of delta 12 fatty acid desaturase from Geotrichum candidum and Mortierella alpina by domain swapping.

Cheese lacks essential fatty acids (EFAs). Delta 12 fatty acid desaturase (FADS12) is a critical enzyme required for EFA biosynthesis in fermentation of the predominant strains of cheese. Previously, we identified the FADS12 gene and characterized its function for the first time in Geotrichum candidum, a dominant strain used to manufacture soft cheese with white rind. In this study, we analyzed the molecular mechanism of FADS12 function by swapping domains from Mortierella alpina and G. candidum that had, respectively, high and low oleic acid conversion rates. The results revealed three regions that are essential to this process, including regions from the end of the second transmembrane domain to the beginning of the third transmembrane domain, from the end of the third transmembrane domain to the beginning of the fourth transmembrane domain, and from the 30-amino acid from the end of the sixth transmembrane domain to the C-terminal end region. Based on our domain swapping analyses, nine pairs of amino acids including H112, S118, H156, Q161, K301, R306, E307, A309 and S323 in MaFADS12 (K123, A129, N167, M172, T302, D307, I308, E310 and D324 in GcFADS12) were identified as having a significantly effect on FADS12 catalytic efficiency, and linoleic acid and its analogues (12,13-cyclopropenoid fatty acid) were found to inhibit the catalytic activity of FADS12 and related recombinant enzymes. Furthermore, the molecular mechanism of FADS12 inhibition was analyzed. The results revealed two allosteric domains, including one domain from the N-terminal region to the beginning of the first transmembrane domain and another from the 31st amino acid from the end of the sixth transmembrane domain to the C terminus. Y4 and F398 amino acid residues from MaFADS12 and eight pairs of amino acids including G56, L60, L344, G10, Q13, S24, K326 and L344 in MaFADS12 (while Y66, F70, F345, F20, Y23, Y34, F327 and F345 in GcFADS12) played a pivotal role in FADS12 inhibition. Finally, we found that both allosteric and active sites were responsible for the catalytic activity of FADS12 at various temperatures, pH, and times. This study offers a solid theoretical basis to develop preconditioning methods to increase the rate at which GcFADS12 converts oleic and linoleic acids to produce higher levels of EFAs in cheese.

Characterization and comparison of metaproteomes in traditional and commercial dajiang, a fermented soybean paste in northeast China.

Dajiang is a popular Chinese fermented soybean condiment. Here, a comparative metaproteomic analysis of traditional and commercial dajiang was performed during fermentation. A total of 4250 and 1421 peptide sequences were obtained from 3493 and 1987 proteins in traditional and commercial dajiang, respectively. 4299 differentially expressed microbial proteins show a high metabolic heterogeneity between the two types of dajiang. The KEGG annotation indicated that there were some pathways related to human diseases, which suggest that some microbes in traditional dajiang fermentation may have greater food safety hazards. In combination with qualitative metabolomic analysis, we further traced metabolic intermediates and key enzymes in several main fermentation pathways of dajiang to be mainly affiliated with Penicillium, Tetracoccus and Bacillus in traditional samples, as well as Aspergilus in commercial samples. These results could provide information for the selection of strains that are more suitable to produce high quality dajiang and other fermented products.

Meta-omics reveal microbial assortments and key enzymes in bean sauce mash, a traditional fermented soybean product.

BACKGROUND: Dajiang is fermented based on the metabolism of microbial communities in bean sauce mash, a traditional fermented soybean product in China. The current study first investigated the metaproteome of bean sauce mash. This was followed by an analysis of its biological functions and its microbial community to reveal information about strains and about the expressed proteins to better understand the roles of the microbiota in bean sauce mash. RESULTS: The metaproteomic results demonstrated that a total of 1415 microbial protein clusters were expressed mainly by members of the Penicillium and Rhizopus genera and were classified into 100 cellular components, 238 biological processes, and 220 molecular function categories by gene ontology (GO) annotation. Enzymes associated with glycolysis metabolic pathways were also identified. These can provide the energy required for microbial fermentation. Illumina MiSeq sequencing technology results showed that the microorganism communities of bean sauce mash exhibited a high level of diversity. Microbiological analysis demonstrated that the Penicillium, Mucor, Fusarium, Aspergillus, and Rhizopus fungi, and Lactobacillus, Enterococcus, Fructobacillus, Staphylococcus, Carnobacterium genera were predominant 22 samples. CONCLUSION: The profiles and insights in the current study are important for research on bean sauce mash and related products in terms of their food microbial ecology. The information obtained from this study will help the development of stable sufu starter cultures with unique sensory qualities. © 2019 Society of Chemical Industry.

An integrated metagenomic/metaproteomic investigation of microbiota in dajiang-meju, a traditional fermented soybean product in Northeast China.

Dajiang-meju have been used as major ingredients for the preparation of traditional spontaneously fermented soybean paste in Northeast China. In this work, we sequenced and analyzed the metagenome of 12 dajiang-meju samples. To complement the metagenome analysis, we analyzed the taxonomic and functional diversity of the microbiota by metaproteomics (LC-MS/MS). The analysis of metagenomic data revealed that the communities were primarily dominated by Enterobacter, Enterococcus, Leuconostoc, Lactobacillus, Citrobacter and Leclercia. Moreover, changes in the functional levels were monitored, and metaproteomic analysis revealed that most of the proteins were mainly expressed by members of Rhizopus, Penicillium and Geotrichum. The number of sequences allocated to fungi in the fermentation process decreased, whereas the number of sequences assigned to bacteria increased with time of fermentation. In addition, functional metagenomic profiling indicated that a series of sequences related to carbohydrates and amino acids metabolism were enriched. Additionally, enzymes associated with glycolysis metabolic pathways were presumed to contribute to the generation of flavor in dajiang-meju. Proteins from different dajiang-meju samples involved in global and overview maps, carbohydrate metabolism, nucleic acid metabolism and energy metabolism were differentially expressed. This information improves the understanding of microbial metabolic patterns with respect to the metaproteomes of dajiang-meju and provides a powerful tool for studying the fermentation process of soybean products.

Metaproteomics of Microbiota in Naturally Fermented Soybean Paste, Da-jiang.

Da-jiang is a typical traditional fermented soybean product in China. At present, the proteins in da-jiang are needed to be explored. The composition and species of microbial proteins in traditional fermented da-jiang were analyzed by metaproteomics based on sodium dodecyl sulfonate-polyacrylamide gel electrophoresis (SDS-PAGE) and liquid chromatography-mass spectrometry/mass spectrometry (LC-MS/MS). The results showed that the number and variety of microbial proteins in the traditional fermented da-jiang from different regions were different. The production site influences the fermentation in da-jiang. Then we analyzed the functions of the microbial proteins identified in da-jiang, and found that they were mainly involved in the process of protein synthesis, glycometabolism and nucleic acid synthesis. In addtion, we compared the proteins composition in different da-jiang. There are 51 common proteins of naturally fermented da-jiang, and 25 common microbial sources. The main commonly microbial sources of fungal proteins are Saccharomyces cerevisiae and Schizosaccharomyces; the main commonly microbial sources of bacterial proteins are Enterococcus faecalis, Leuconostoc mesenteroides, Acinetobacter baumannii, and Bacillus subtilis. These common microbes play the predominant role in da-jiang fermentation. The present results help us to understand the fermentation of da-jiang and improve the quality and safety of final products in the future. PRACTICAL APPLICATION: The study illustrated metaproteome of microbiota in traditional fermented soybean paste, da-jiang, by sodium dodecyl sulfonate-polyacrylamide gel electrophoresis (SDS-PAGE) and liquid chromatography-mass spectrometry/mass spectrometry (LC-MS/MS). A method of extracting metaproteome from microbiota in da-jiang was attempted. The findings help to understand the fermentation of da-jiang and improve the quality and safety of da-jiang in fermented industry.