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The importance of nonconventional yeasts has increasingly been highlighted, particularly for aroma formation in fermented foods. Here, we performed de novo whole-genome sequencing of Wickerhamomyces subpelliculosus , which produces a variety of volatile flavor compounds, leading to the identification of the alcohol acyltransferase (AATase) family of genes. The genome of W. subpelliculosus contains seven AATase genes, encoding alcohol- O -acetyltransferases (ATFs) and ethanol acetyltransferase 1 (EAT1) for acetate ester formation, along with ethanol hexanoyl transferase 1 (EHT1) for ethyl ester formation. Among five ATF homologues, only WsATF5 showed acetyltransferase activity toward myriocin, a structural analogue of sphingosine. In contrast, heterologous expression of WsEHT1 and WsEAT1 in Saccharomyces cerevisiae promoted the production of ethyl decanoate and ethyl acetate, respectively, supporting their AATase activity. The enzymatic activity analyses revealed the additional alcoholysis activity of WsEAT1 and the thioesterase activity of WsEHT1. Subcellular localization analysis indicated that WsEAT1 was localized in the mitochondria, WsEHT1 in the endoplasmic reticulum and lipid droplets (LDs), and WsATF5 in the LDs. The novel W. subpelliculosus AATases could be usefully applied to produce flavor components in various food industries.

N -Lactoyl-phenylalanine (Lac-Phe) is a metabolite known for its appetite-suppressing and antiobesity properties, while phenyllactic acid (PLA) is recognized for its antibacterial activity. Both metabolites are derived from phenylalanine and lactic acid metabolism through peptidase and dehydrogenase activities. The aim of this study was to investigate the production of Lac-Phe and PLA in kimchi, focusing on the role of lactic acid bacteria (LAB). Ultrahigh performance liquid chromatography coupled with time-of-flight mass spectrometry was used to quantify these metabolites in homemade and commercial kimchi. Lac-Phe and PLA were detected in both kimchi sample types. Various genera, including Lactobacillus , Leuconostoc , and Weissella, were evaluated for Lac-Phe and PLA production. LAB strains exhibiting high Lac-Phe production generally showed lower PLA production, indicating an inverse relationship between these two metabolites. Analysis of dipeptidase sequences revealed that the presence of carnosine dipeptidase 2 (CNDP2)-like M20 metallopeptidase is crucial for Lac-Phe production in LAB. Additionally, phenylalanine was identified as a major factor for both Lac-Phe and PLA production, whereas lactic acid supplementation did not significantly affect their production levels.

The diet in early life is essential for the growth and intestinal health later in life. However, beneficial effects of a diet enriched in branched short-chain fatty acids (BSCFAs) for infants are ambiguous. This study aimed to develop a novel fermented protein food, enriched with BSCFAs and assess the effects of dry and wet ferment products on young pig development, nutrient absorption, intestinal barrier function, and gut microbiota and metabolites. A total of 18 young pigs were randomly assigned to three groups. The dry corn gluten-wheat bran mixture (DFCGW) and wet corn gluten-wheat bran mixture (WFCGW) were utilized as replacements for 10% soybean meal in the basal diet. Our results exhibited that the WFCGW diet significantly increased the growth performance of young pigs, enhanced the expression of tight junction proteins, and regulated associated cytokines expression in the colonic mucosa. Simultaneously, the WFCGW diet led to elevated levels of colonic isobutyric and isovaleric acid, as well as the activation of GPR41 and GPR109A. Furthermore, more potential probiotics including Lactobacillus , Megasphaera , and Lachnospiraceae_ND3007_group were enriched in the WFCGW group and positively associated with the beneficial metabolites such as 5-hydroxyindole-3-acetic acid. Differential metabolite KEGG pathway analysis suggested that WFCGW might exert gut health benefits by modulating tryptophan metabolism. In addition, the WFCGW diet significantly increased ghrelin concentrations in serum and hypothalamus and promoted the appetite of young pigs by activating hypothalamic NPY/AGRP neurons. This study extends the knowledge of BSCFAs and provides a reference for the fermented food application in the infant diet.

The control of excess biogenic amines (BAs) is crucial for the sustainable development of fermented foods. This study aimed to screen endogenous functional strains in Doubanjiang with the capacity to degrade BAs and to elucidate their application potential. Pediococcus acidilactici L-9 (PA), which was confirmed as a safe strain by phenotypic and genotypic analyses, exhibited an efficient degradation ability on BAs, particularly regarding tyramine. Notably, the degradation of tyramine was maintained at 24.03-50.60% at different temperatures (20-40 °C), pH values (4.0-9.0), and NaCl concentrations (3-18%, w/v). Additionally, genomic data revealed the presence of the laccase-coding gene, which was demonstrated to play a pivotal role in BA degradation by heterologous expression. Further, molecular docking results indicated that the degradation of BA by laccase is closely linked to the electron transfer pathway formed by the substrate and key amino acid residues. Finally, the degradation of tyramine by PA remained within the range of 8.19-64.19% under the simulated system with 6-12% salinity. This study provided valuable insights into the safety of PA and its potential degradation capacity on BAs, particularly in mitigating tyramine accumulation, which could improve the quality of Doubanjiang and other fermented foods.

Due to the reports describing virulent and multidrug resistant enterococci, their use has become a topic of controversy despite most of them being safe and commonly used in traditionally fermented foods worldwide. We have characterized Enterococcus lactis SF68, a probiotic strain approved by the European Food Safety Authority (EFSA) for use in food and feed, and find that it has a remarkable potential in food fermentations. Genome analysis revealed the potential of SF68 to metabolize a multitude of carbohydrates, including lactose and sucrose, which was substantiated experimentally. Bacteriocin biosynthesis clusters were identified and SF68 was found to display a strong inhibitory effect against Listeria monocytogenes . Fermentation-wise, E. lactis SF68 was remarkably like Lactococcus lactis and displayed a clear mixed-acid shift on slowly fermented sugars. SF68 could produce the butter aroma compounds, acetoin and diacetyl, the production of which was enhanced under aerated conditions in a strain deficient in lactate dehydrogenase activity. Overall, E. lactis SF68 was found to be versatile, with a broad carbohydrate utilization capacity, a capacity for producing bacteriocins, and an ability to grow at elevated temperatures. This is key to eliminating pathogenic and spoilage microorganisms that are frequently associated with fermented foods.

Lactobacilli fermentation possesses special nutritional and health values to food, especially in improving diseases related to the gut microbiota such as osteoporosis risk. Previous research indicates that lactobacilli-fermented foods have the potential to enhance the bone mineral density (BMD), as suggested by some clinical studies. Nonetheless, there is currently a lack of comprehensive summaries of the effects and potential mechanisms of lactobacilli-fermented foods on BMD. This review summarizes findings from preclinical and clinical studies, revealing that lactobacilli possess the potential to mitigate age-related and secondary factor-induced bone loss. Furthermore, these findings imply that lactobacilli are likely mediated through the modulation of bone remodeling via gut inflammation-related pathways. Additionally, lactobacilli fermentation may augment calcium accessibility through directly promoting calcium absorption or modifying food constituents. Considering the escalating global health challenge of bone-related issues among the elderly population, this review may offer a valuable reference for the development of food strategies aimed at preventing osteoporosis.

Elevated levels of biogenic amines (BAs) in fermented food can have negative effects on both the flavor and health. Mining enzymes that degrade BAs is an effective strategy for controlling their content. The study screened a strain of Lactobacillus hilgardii 1614 from fermented food system that can degrade BAs. The multiple copper oxidase genes LHMCO 1614 were successfully mined after the whole genome protein sequences of homologous strains were clustered and followed by homology modeling. The enzyme molecules can interact with BAs to stabilize composite structures for catalytic degradation, as shown by molecular docking results. Ingeniously, the kinetic data showed that purified LHMCO 1614 was less sensitive to the substrate inhibition of tyramine and phenylethylamine. The degradation rates of tyramine and phenylethylamine in huangjiu (18% vol) after adding LHMCO 1614 were 41.35 and 40.21%, respectively. Furthermore, LHMCO 1614 demonstrated universality in degrading tyramine and phenylethylamine present in other fermented foods as well. HS-SPME-GC-MS analysis revealed that, except for aldehydes, the addition of enzyme treatment did not significantly alter the levels of major flavor compounds in enzymatically treated fermented foods ( p > 0.05). This study presents an enzymatic approach for regulating tyramine and phenylethylamine levels in fermented foods with potential applications both targeted and universal.

As a traditional Thai condiment, Pla-ra is used to add flavor and richness to dishes. Nine treatment combinations of Pla-ra formulations created from 3 types of fish (Mor fish, Kradee fish, and Mor + Kradee fish) and 4 different carbohydrate sources (none, rice bran, roasted rice, and rice bran─roasted rice mixture) were studied through a 12 month fermentation period (1, 3, 5, 7, 8, 9, 10, 11, and 12 months). 16S rRNA Next Generation Sequencing (NGS) and LC-MS/MS techniques were used to analyze the microbial diversity and identify taste-enhancing peptides. Descriptive sensory analysis was performed on the extracts of the 108 Pla-ra samples mixed in a model broth. Koku perception and saltiness-enhancing attributes were clearly perceived and dominant in all samples, even though glutamyl peptides, including γ-Glu-Val-Gly, were found at subthreshold levels. The samples from mixed fish and Mor fish fermented with roasted ground rice and rice bran for 12 months had the most typical Pla-ra odors and tastes and had high taste-enhancing activities. NGS analysis revealed the presence of bacteria containing a large number of protease and aminopeptidase genes in the samples. Bacillus spp., Gallicola spp., and Proteiniclasticum spp. correlated well with the generation of glutamyl and arginyl peptides and typical odors in the samples. These results confirmed the typical sensory quality of Pla-ra depended on protein sources, carbohydrate sources, and bacteria communities. Further optimization of the microbial composition found could lead to the development of starter cultures to control and promote flavor development in fermented fish products.

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.

Doenjang (fermented soybean paste) and ganjang (soy sauce) are traditional fermented foods that are widely consumed in Korea. The oligosaccharides found in soybean and its fermented foods have great potential to improve the quality of foods; however, their structural details have not been well studied. In this study, we used advanced mass spectrometry and gas chromatography to analyze oligosaccharides and their monomeric composition in two fermented soybean products. In both foods, oligosaccharides with a degree of polymerization ranging from 3 to 7 were found. Their constituent monosaccharides were characterized; galactose, xylose, arabinose, and rhamnose were the predominant constituents of the oligosaccharides, and fucose, fructose, mannose, glucose, and N -acetylglucosamine were also found. The great structural diversity of the oligosaccharides found suggests that soybean carbohydrates are hydrolyzed and/or transformed during fermentation and may yield novel oligosaccharides.

The aim of this study was to isolate the angiotensin I-converting enzyme (ACE) inhibitory peptides from the skirt of Chlamys farreri fermented with Bacillus natto and to explore the antihypertension effect through in vivo studies. ACE inhibitory peptides were purified from the fermentation mixture by ultrafiltration, gel filtration chromatography, and reversed-phase high-performance liquid chromatography sequentially. The amino acids' sequence of the five novel ACE inhibitory peptides were identified by liquid chromatography-tandem mass spectrometry. Animal experiments demonstrated that the novel ACE inhibitory peptides significantly reduced the blood pressure in spontaneously hypertensive rats after a single or long-time treatment. Potential mechanisms were explored, and the results indicated that the novel peptides could regulate the renal renin-angiotensin system, improve vascular remodeling, inhibit myocardial fibrosis, and rebalance the gut microbial dysbiosis. Our results suggest that the fermentation products of the Chlamys farreri skirt by B. natto are potential sources of active peptides processing antihypertension activities.

Application of gas chromatography-olfactometry (GC-O) analysis to pumpernickel bread and its intermediate products obtained during the fermentative process allowed for the recognition of 32 key aroma compounds with specific odor notes. Subsequent quantitation using the stable isotope dilution assay (SIDA) and calculation of odor activity values (OAVs) revealed 22 compounds with OAV > 1, with the highest OAV for 3-methylbutanal with a malty odor (6660), 2-methylbutanal with a malty odor (4560), 3-(methylthio)propanal with a boiled potato odor (2047), 3-hydroxy-4,5-dimethyl-2(5 H )-furanone with a lovage-like odor (1233), dimethyl trisulfide with a cabbage-like odor (475), 2-phenylethanol with a rosy odor (414), β-damascenone with a boiled apple odor (200), 4-hydroxy-3-methoxybenzaldehyde with a vanilla-like odor (184), 4-hydroxy-2,5-dimethyl-3(2 H )-furanone with a caramel odor (172), and phenylacetladehyde with a honey odor (100). The results showed that aroma of pumpernickel bread is mostly formed during baking; however, the fermentation process influences the formation of several compounds, such as ethyl acetate, 2,3-butanedione, acetic acid, 3-methyl-1-butanol, ethyl butyrate, ethyl lactate, ethyl-3-methyl butyrate, ethyl hexanoate, phenylacetaldehyde, and 2-phenylethanol. Furthermore, the addition of functional components, such as malt and wheat fiber, affects the concentration of several key odorants in the final product, such as 2- and 3-methylbutanals, 3-hydroxy-2-methyl-4 H -pyran-4-one, and 4-hydroxy-3-methoxybenzaldehyde.

A 16S rRNA next-generation sequencing technique was applied to investigate the microbial diversity and liquid chromatography-tandem mass spectrometry was used to identify glutamyl peptide profiles of 10 Thai fermented freshwater fish (Pla-ra) samples. A total of 12 genera of bacteria were able to be detected, with Tetragenococcus spp., Staphylococcus spp., and Lactobacillus spp. dominating. Of the 18 glutamyl peptides analyzed, 17 were found, even though the amounts detected were lower than the taste threshold. Despite this, an increase in mouthfulness sensation, reflecting kokumi activity, was clearly identified in most of the samples, which might be because of a synergistic effect of different sub-threshold compounds present in the samples. In principle component analysis, the relationship between microorganisms and glutamyl peptide generation was observed, especially between Tetragenococcus spp. and Lentibacillus spp. and the generation of γ-Glu-Val-Gly. Correlations between microbial diversity and the generation of taste enhancers were identified in this study.

Fermented foods have unique microbiota and metabolomic profiles that can support dietary diversity, digestion, and gut health of consumers. Laowo ham (LWH) is an example of an indigenous fermented food from Southwestern China that has cultural, ecological, economic, and health significance to local communities. We carried out ethnobiological surveys coupled with metagenomic and metabolomic analyses using GC-TOF-MS and UPLC-QTOF-MS to elucidate the microbiota and metabolic profiles of LWH samples at different ripening stages. The results from high-throughput sequencing showed a total of 502 bacterial genera in LWH samples with 12 genera of bacteria and 6 genera of fungi identified as dominant groups. This is the first study to our knowledge to report the bacteria of Lentibacillus and Mesorhizobium along with fungi Eremascus and Xerochrysium on a fermented meat product. Findings further revealed that the metabolite profiles among LWH samples were significantly different. In total, 27 and 30 metabolites from GC-TOF-MS and UPLC-QTOF-MS analysis, respectively, were annotated as highly discriminative metabolites. Among the differential compounds, the relative contents of most amino acids showed the highest in the LWH sample ripened for two years, while some metabolites with potential therapeutic effects such as levetiracetam were the most abundant in the LWH sample ripened for three years. The correlation analysis indicated that the dominant microbes were closely related to differential metabolites, highlighting the importance of their functional characterization. Findings indicate that the consumption of LWH contributes to microbiological and chemical diversity of human diets as well as suggests efficacy of combining GC-MS and LC-MS to study the metabolites in dry-cured meat products.

Melatonin has recently been detected in fermented beverages and foods, in which microorganism metabolism is highly important. The existing literature knowledge discusses the direction for future studies in this review. Evidence shows that many species of microorganisms could synthesize melatonin. However, the actual concentrations of melatonin in fermented foods and beverages range from picograms per milliliter to nanograms per milliliter. Different types of microorganisms, different raw materials, different culture environments, the presence or absence of precursors, high or low alcohol content, and different detection methods are all possible reasons for the huge difference of melatonin levels. Thus far, there have been relatively few studies on the melatonin synthesis pathway microorganisms. Thus, referring to the synthetic pathway of plants and animals, the putative melatonin biosynthesis pathway of microorganisms is presented. It will be significant to discuss whether all species of microorganisms have the capacity to synthesize melatonin and what the biological functions of melatonin are in microorganisms. Melatonin plays a lot of important roles in microorganisms, particularly in enhancing the tolerance of environment stress. Also, the loss of melatonin concentration in commercially available fermented foods and beverages is a ubiquitous trend, and how to solve this problem is a new field to be further explored.

Biogenic amines (BAs) are low molecular weight organic bases. BAs occurring naturally in living organisms are responsible for a number of vital functions, including (in humans) secretion of gastric acids, controlling body temperature, differentiation and growth of cells, immune reactions, and brain activity. However, if oversupplied with food, BAs may cause food poisoning and produce undesirable effects. Nine BAs and eight free amino acids (FAAs) were determined in 85 samples of 19 different varieties of fermented vegetables available on the Polish retail market. Both BA and FAA levels differed significantly among various varieties of the studied fermented vegetables. Averages for the sum of all tested BAs ranged from 30.29 ± 16.43 mg·kg -1 in fermented olives to 612.1 ± 359.33 mg·kg -1 in fermented Brussels sprout. BA profiles were dominated by putrescine (42%), tyramine (20%), cadaverine (18%), and histamine (8%); jointly, the four amines amounted to 88% of all nine studied BAs. The combined level of the latter four BAs was calculated for each vegetable variety as the so-called BA index (BAI). On that basis, the risk of BA-related adverse health effects has been assessed as high/medium/low in 6/3/10 of all 19 studied varieties of fermented vegetables. Brussels sprout and broccoli turned out to be the most risky vegetables from that point of view (BAI above 400 mg·kg -1 ). FAA levels ranged from 54.8 ± 12.76 (fermented olives) to 3917.42 ± 1528.73 mg·kg -1 (fermented garlic). The high content of FAAs may increase the risk of forming toxic amounts of BAs, depending on characteristics of the current and added microflora as well as on environmental and technological conditions the product is subjected to.

Fermented black garlic has multiple beneficial biological activities, including cardiovascular protection, anticancer, hepatoprotective, and antibacterial properties. In this study, metabolic differences in the properties of black and fresh garlic were investigated via liquid chromatography quadrupole/time-of-flight-based metabolomics, leading to the identification of characteristic components. Fermented black garlic samples and their Amadori products (AC) promoted angiogenesis, prevented thrombus formation by rescuing chemical-induced vascular lesions in zebrafish, and inhibited H 2 O 2 -induced injury of endothelial cells, thus reducing the risk of cardiovascular disease. AC suppressed activation of the mitogen-activated protein kinase pathway through inhibition of p38 and ERK1/2 phosphorylation, in turn, increasing the availability of c-Fos/c-Jun or c-Jun/c-Jun complexes for apoptotic resistance. Clarification of the associated signaling pathways should therefore provide a solid foundation for optimization of black garlic-based therapies.

The ability of Debaryomyces hansenii to produce volatile sulfur compounds from sulfur amino acids and the metabolic pathway involved have been studied in seven strains from different food origins. Our results proved that l-methionine is the main precursor for sulfur compound generation. Crucial differences in the sulfur compound profile and amino acid consumption among D. hansenii strains isolated from different food sources were observed. Strains isolated from dry pork sausages displayed the most complex sulfur compound profiles. Sulfur compound production, such as that of methional, could result from chemical reactions or yeast metabolism, while according to this study, thioester methyl thioacetate appeared to be generated by yeast metabolism. No relationship between sulfur compounds production by D. hansenii strains and the expression of genes involved in sulfur amino acid metabolism was found, except for the ATF2 gene in the L1 strain for production of methyl thioacetate. Our results suggest a complex scenario during sulfur compound production by D. hansenii .

Aiming at the identification of the key bitter peptides in fermented foods, a new approach, coined "sensoproteomics", was developed and applied to fresh cheese samples differing in bitter taste intensity. By means of MPLC fractionation of the water-soluble cheese extracts in combination with taste dilution analysis, complex fractions with intense bitter taste were located and then screened by UPLC-MS/MS for the entire repertoire of ∼1600 candidate peptides, extracted from a literature meta-analysis on dairy products, by using a total of 120 selected reaction monitoring methods computed in silico. A total of 340 out of the 1600 peptides were found in the cheese samples, among which 17 peptides were identified as candidate bitter peptides by considering only peptides that were located in the bitter-tasting MPLC fractions (signal-to-noise ratio: ≥10) with a fold-change of ≥3 when comparing the less bitter to the more bitter cheese sample and that were validated by comparison with the synthetic reference peptides. While EIVPNS[phos]VEQK (α s1 -CN 70-78 ) and INTIASGEPT (κ-CN 122-131 ) did not exhibit any bitter taste up to 2000 μmol/L, 15 of the 17 target peptides showed bitter taste thresholds ranging from 30 (ARHPHPHLSFM, κ-CN 96-106 ) to 690 μmol/L (IQKEDVPS, α s1 -CN 81-88 ). Finally, quantitative peptide analysis followed by calculation of dose-overthreshold factors revealed a primary contribution of MAPKHKEMPFPKYPVEPF (β-CN 102-119 ) and ARHPHPHLSFM (κ-CN 96-106 ) to the perceived bitter taste of the fresh cheese samples. Finally, the evolution of the bitter peptides throughout two different fresh cheese manufacturing processes was quantitatively recorded.

This study investigated the inhibitory effects of kimchi bioactive compounds against endoplasmic reticulum (ER) stress-induced apoptosis in amyloid beta (Aβ)-injected mice. Mice received a single intracerebroventricular injection of Aβ 25-35 , except for the normal group. Mice were subjected to oral administration of 10 mg of capsaicin, 50 mg of 3-(4'-hydroxyl-3',5'-dimethoxyphenyl)propionic acid (HDMPPA), 50 mg of quercetin, 50 mg of ascorbic acid, or 200 mg of kimchi methanol extract (KME) per kilogram of body weight for 2 weeks ( n = 7 per group). In the in vitro blood-brain barrier (BBB) permeability test, all bioactive compounds penetrated the BBB except ascorbic acid. The protein expression level of APP, BACE, and p-Tau elevated by Aβ injection was decreased by kimchi bioactive compounds ( P < 0.05). Quercetin, HDMPPA, and KME decreased oxidative stress, as indicated by ROS and TBARS levels ( P < 0.05). The protein expression level of ER stress markers GRP78, p-PERK, p-eIF2α, XBP1, and CHOP and the proapoptotic molecules Bax, p-JNK, and cleaved caspases-3 and -9 decreased ( P < 0.05). In contrast, the protein expression level of antiapoptotic molecules Bcl2 and cIAP increased ( P < 0.05). These results were supported by histological analysis.