Search Results (104)

Processed meat products represent a major challenge for proteomic species identification due to extensive thermal treatment and protein structural changes. In this study, species-specific peptides in pork, chicken, and bovine meat products were identified using a directed fragmentation-assisted de novo sequencing workflow that combines 4-formylbenzene-1,3-disulfonic acid (FBDA) peptide derivatization, dual-polarity data-independent mass spectrometry (DIA-MS), and Protein Acrobat de novo sequencing software. Comparative analysis of non-fractionated and strong cation exchange (SCX)-fractionated pork luncheon samples improved peptide and protein identification after fractionation, with 312 peptides and 115 protein groups detected exclusively in fractionated samples. Species-specific peptides were predominantly assigned to conserved muscle-related proteins, including myosin, troponin, and tropomyosin, while sequence variability enabled reliable species discrimination despite protein conservation across species. To evaluate applicability for food fraud detection, mixed meat samples containing 10% chicken in pork or bovine matrices were analyzed, reflecting potential economically motivated adulteration through substitution with lower-cost meat components. Several chicken-specific peptides remained detectable in both mixtures, demonstrating robustness of the FBDA-assisted peptide sequencing combined with SCX fractionation and DIA-MS for detection of adulteration in complex processed food matrices. These findings establish a mass spectrometry-driven orthogonal method to ELISA testing for fast, reliable and accurate metaproteome analysis of highly processed food. Full article

As global ecosystems and food systems face unprecedented anthropogenic and climatic challenges, there is a demand for an integrated understanding of biological systems. Proteomics has emerged as a definitive approach offering a direct view of the molecular phenotype, yet it is traditionally separated into plant and animal disciplines. With recent advances in mass spectrometry (MS) and bioinformatics tools, this prospective review proposes that combining a One Health proteomics approach with deep-learning data analysis can revolutionize global food security, animal productivity, and ecosystem health by uncovering proteoform signatures that drive resilience across life. The potential of a unified One Health proteomic framework, highlighting major developments, including 4D proteomics, Data-Independent Acquisition (DIA), and single-cell resolution, and emphasizes their capacity to resolve the complex proteoform landscape across kingdoms. Review emphasizes the applications of proteogenomics as a cross-disciplinary tool to improve genome annotations, explain evolutionary differences, discover biomarkers in animals and resolve complex signaling networks in plants under stress. Nevertheless, contemporary proteogenomics methods still show limitations in their ability to comprehensively resolve proteoforms due to the fact that the use of peptide-based approaches makes it difficult to fully appreciate the post-translational modifications specific to each protein isoform. We show that One Health proteomics will provide a transformative roadmap for deciphering the functional proteoform signatures that underpin resilience across the tree of life. Full article

Seasonal fluctuations in open solid-state fermentation drive batch-to-batch variability in Chinese Baijiu Daqu; however, how environmental shifts reshape microbial functional expression and non-volatile flavour precursors in medium-high-temperature Daqu remains poorly resolved. In this study, data-independent acquisition (DIA)-based quantitative metaproteomics and untargeted liquid chromatography–mass spectrometry (LC-MS) metabolomics were integrated to characterise winter and summer Daqu from Luzhou, Sichuan. Among 2904 annotated non-volatile metabolites, orthogonal partial least squares discriminant analysis (OPLS-DA) revealed clear seasonal separation; 1472 differential metabolites (560 up- and 912 downregulated in winter vs. summer; variable importance in projection [VIP] > 1, p < 0.05) were enriched in glycolysis/gluconeogenesis, the tricarboxylic acid (TCA) cycle, amino acid biosynthesis, and starch/sucrose metabolism. DIA-based quantitative metaproteomics further resolved season-specific enzyme expression: summer Daqu exhibited elevated saccharolytic, glycolytic and amino-acid-converting enzymes (β-glucosidase, 6-phosphofructokinase, pyruvate dehydrogenase), whereas winter Daqu was enriched in glucose oxidase, phosphoenolpyruvate carboxykinase and aldehyde dehydrogenase, consistent with a pattern suggestive of carbon-storage prioritisation. Proteome–metabolome integration established a coherent “enzyme protein abundance–inferred metabolic tendency–metabolite accumulation” correlative framework axis: higher hydrolytic and central-carbon enzyme abundance in summer corresponded to increased maltose, lactate, acetate, L-glutamate and L-aspartate. Therefore, production season reshapes Daqu quality chiefly by corresponding to distinct patterns of in situ enzyme protein abundance, providing a DIA quantitative metaproteome-anchored mechanistic framework for screening high-expression starters and stabilising seasonal Daqu quality. Full article

Background: Acetification is a complex process driven by acetic acid bacteria (AAB), in which high ethanol and acidity levels require strong microbial metabolic adaptation. Although the microbiota involved in vinegar production has been described, the functional mechanisms that enable these bacteria to maintain metabolic activity remain poorly understood. In this study, the functional dynamics of AAB during Verdejo vinegar acetification were analyzed using a quantitative metaproteomic approach. Methods: Acetification was performed in submerged culture under semi-continuous conditions, and samples were collected at four stages of the cycle (S1–S4). Results: LC-MS/MS analysis led to the identification of 1626 proteins, of which 1409 were assigned to the Acetobacteraceae family. Komagataeibacter europaeus was the dominant species (73.7%). Hierarchical clustering revealed four protein abundance patterns, and differential analysis identified 350 proteins with increased abundance and 169 with decreased abundance, with the greatest changes observed between S1 and S4. Functional annotation and protein–protein interaction analyses indicated that the main metabolic adaptations involve pathways related to energy metabolism, amino acid biosynthesis, membrane-associated functions, cellular homeostasis, and acid stress response. Conclusions: Overall, the results show that K. europaeus concentrates most of the metabolic activity during acetification and that proteome reorganization reflects key molecular strategies for adaptation and survival under high-acidity conditions. Full article

Traditional fermented foods and beverages (TFFB) rely on complex microbial communities that generate distinctive flavors, nutritional attributes, and cultural value, but spontaneous or empirically controlled fermentations often limit reproducibility. Defined microbial consortia (DMCs) provide a promising route for improving fermentation controllability and product consistency, although overly simplified starters may fail to reproduce the ecological robustness and sensory complexity of traditional systems. This review focuses on how multi-omics and culturomics can support rational DMC design in TFFB. We summarize how metagenomics, metatranscriptomics, metaproteomics, metabolomics, and culturomics reveal community structure, functional potential, active expression, metabolic output, and cultivable strain resources. Particular attention is given to translating multi-omics evidence into strain prioritization through the identification of keystone microorganisms that drive core fermentation functions and helper microorganisms that support ecological or metabolic stability. We further propose an Assembly-Assessment-Redesign (A-A-R) framework for iterative DMC optimization, linking strain selection, functional validation, performance evaluation, and consortium redesign. Finally, we discuss key challenges, including cross-omics integration, experimental verification of microbial functions, standardized validation criteria, and the transfer of laboratory-designed consortia to industrial fermentation systems. Full article

The bioconversion of propionate, a well-known intermediate of anaerobic digestion (AD), to methane is energetically unfavorable under standard conditions, which typically occurs in the syntrophy of bacteria and methanogens via methylmalonyl-CoA (MMC) and the dismutation pathway. Since the latter, which is reported only in Smithella, possessed a thermodynamic advantage over the former, enriching Smithella and promoting the syntrophic interaction and metabolism of the microbiota are important for improving AD efficiency. In this study, lysine-modified Fe₃O₄ (Lys@Fe₃O₄) significantly enhanced the bioconversion of propionate to methane. The methane yield and the maximum methane production rate (R_max) in a Lys@Fe₃O₄ reactor were 278.7% and 271.7% of Blank, and the corresponding values were 201.9% and 201.6% of bare Fe₃O₄, respectively. The metaproteomic results indicated that Lys@Fe₃O₄ increased not only the abundance of Smithella but also the expression of cell surface and adhesion proteins, thereby promoting syntrophic interaction between Smithella and methanogens and facilitating electron and acetate transfer from Smithella to methanogens. Moreover, the expression of quorum-sensing proteins was enhanced, benefiting the cooperation of Smithella and its associated bacterium (Syntrophomonas). Furthermore, the expressions of key enzymes related to metabolism and electron transfer in propionate oxidation, butyrate oxidation, CO₂-reductive methanogenesis and acetoclastic methanogenesis were all significantly upregulated. The results are of great significance for maintaining low propionate concentration and stability of AD. Full article

The production of Chinese Baijiu relies on the synergistic metabolism of multi-species microbial communities in an open environment. Its intricate microbial succession and flavor formation mechanisms have long been considered complex systems that are difficult to fully deconstruct. Traditional culture-dependent techniques inherently fail to comprehensively capture the actual functional roles and dynamic regulation of “viable but non-culturable” (VBNC) microorganisms within this complex system. In recent years, the rapid advancement of multi-omics technologies has offered a novel perspective for elucidating the underlying fermentation mechanisms of Baijiu. This paper systematically reviews the recent progress in the application of metagenomics, metatranscriptomics, metaproteomics, and metabolomics in Baijiu research. Specific focus is placed on the unique contributions of these tools to resolving microbial community structural diversity, mining key functional genes and enzymes, uncovering microbial stress response mechanisms under environmental fluctuations, identifying phages and spoilage microorganisms, and tracing the metabolic pathways of flavor substances. Furthermore, the pivotal role of multi-omics integration strategies in constructing “microbe–metabolite” regulatory networks is highlighted. Finally, current challenges regarding standardization and data integration are discussed, with an outlook on leveraging omics big data to promote digital monitoring and intelligent brewing in the Baijiu industry. Full article

Interactions among plant roots, soil, and microorganisms in the rhizosphere regulate nutrient cycling, plant health, and ecosystem resilience. Recent advances in DNA sequencing and multi-omics are contributing to a shift from primarily descriptive surveys toward more mechanistic and predictive frameworks. This review synthesizes methodological developments and conceptual insights spanning microbial ecology, functional genomics, and agricultural applications. We first summarize DNA-based approaches—marker-gene sequencing, shotgun metagenomics, and quantitative nucleic acid assays—and then complementary omics layers, including metatranscriptomics, metaproteomics, metabolomics, epigenomics, ionomics, and phenomics. We next outline computational advances in data integration, network modeling, and visualization that help represent complex multi-layered datasets as biologically interpretable systems. Applications relevant to climate resilience and sustainable agriculture are discussed, including the design of synthetic microbial communities, the identification of biomarkers for soil health and stress tolerance, and case studies in which rhizosphere multi-omics informs crop breeding and soil management strategies. Overall, these developments underscore the potential of treating microbes as functional and, to some extent, manageable components of the plant holobiont. Looking ahead, we identify key research gaps involving standardized workflows, cross-scale causal inference, and real-time monitoring pipelines that integrate molecular diagnostics with remote sensing and edge–cloud analytics. By linking ecological mechanisms with translational practice, multi-omics frameworks may support the development of more sustainable, data-driven agriculture that better aligns productivity with environmental stewardship. Full article

Brown leaf spot, a severe fungal disease in Rhododendron, substantially impairs its ornamental and economic value. However, the molecular mechanisms governing the interaction between Rhododendron leaves and the pathogen are still poorly understood. In this study, the less susceptible Rhododendron hybridum ‘Yangmeihong’ (YMH) and the highly susceptible R. hybridum ‘Xiaotaohong’ (XTH) leaves were selected for transcriptomic and widely targeted metabolomic analyses. We compared constitutive metabolite profiles prior to inoculation as well as differentially expressed gene (DEGs) and differentially accumulated metabolites (DAMs) at two days post-inoculation (dpi) with Neopestalotiopsis clavispora. Results showed that YMH leaves exhibited 273 upregulated constitutive metabolites, whereas XTH had only 46. After infection, DEGs in YMH leaves were significantly enriched in pathways including flavonoid biosynthesis and glycerolipid metabolism, while XTH showed enrichment in vitamin B6 metabolism and glutathione metabolism. Metabolomic analysis identified 788 metabolites at 2 dpi, with YMH accumulating 49 DAMs and XTH accumulating 115 DAMs. Integrated transcriptomic and metabolomic analyses further revealed distinctions in reactive oxygen species (ROS) scavenging mechanisms between the two cultivars. These findings offer valuable insights for developing disease-resistant breeding and control strategies in Rhododendron. Full article

In order to understand the dynamic interaction process among species, enzymes, and metabolites during the fermentation process of Baobaoqu, which is a representative Daqu starter for Chinese baijiu, the intimate connection between the progression of microbial communities and the diversities and activities of enzymes was examined by metagenomics, metatranscriptomics and metaproteomics. It was found that while 5211 species of microorganisms were detected by metagenomics, only 1774 active species were detected by metatranscriptomics, which indicated that only a small proportion (34.04%) were active. The metabolic routes associated with the breakdown of substrates and synthesis of metabolites were redesigned, and the special functional microorganisms for lactate, pyrazines and phenylethyl alcohol production were isolated. It was found that the progression of the microbial community was highly coupled with the components of enzymes and flavor substrates, precisely corresponding to the three stages of the Baobaoqu fermentation process, and were regulated by multiple physical factors. During the Baobaoqu-making process of the fermentation, microorganisms with different functions work together to complete metabolism in different stages. These findings will aid us in gaining a deeper and clearer understanding of the “species–enzyme–metabolite” system within the Daqu starter culture, thus offering valuable perspectives for developing artificial synthetic communities and the production of high-quality Baobaoqu. Full article

Although vinegar is technically elaborated by a well-known bioprocess, the behavior and function of the microorganisms responsible for its production still need investigation. In vinegars obtained from raw materials and systems typical of Europe, the acetic acid bacteria species Komagataeibacter europaeus predominates due to its particular adaptive metabolism. This work addresses the study of several adaptation mechanisms of K. europaeus during acetic acid fermentation in a submerged semi-continuous production system. The aim is to analyze the molecular response and behavior of this species to increasing acidity gradients, up to 7–8% w/v acetic acid, applying a comparative proteomic approach in three matrices (synthetic alcoholic medium, dark craft beer, and dry fine wine). A total of 1070 proteins are identified, with 174 showing statistically significant changes in abundance (FDR < 0.05), particularly in pathways related to amino acid biosynthesis, fatty acid metabolism, and stress response. The proteomic patterns differ among substrates, with the synthetic alcohol medium inducing stress-related proteins and the dark craft beer enhancing lipid biosynthesis. These observations provide experimental evidence that the fermentation substrate modulates metabolic adaptation in K. europaeus, offering a rational basis for designing fermentation protocols that enhance bacterial resilience, thereby optimizing vinegar production processes. Full article

The human skin microbiota, a complex community of bacterial, fungal, and viral organisms, plays a crucial role in maintaining skin homeostasis and regulating host-pathogen interactions. Dysbiosis within this microbial ecosystem has been implicated in various dermatological conditions, including acne vulgaris, psoriasis, seborrheic dermatitis, and atopic dermatitis. This review, for the first time, provides recent advancements in all four layers of omic technologies—metagenomics, metatranscriptomics, metaproteomics, and metabolomics—offering comprehensive insights into microbial diversity, in the context of functional skin modeling. Thus, this review explores the application of these omic tools to in vitro skin models, providing an integrated framework for understanding the molecular mechanisms underlying skin–microbiota interactions in both healthy and pathological contexts. We highlight the importance of developing advanced in vitro skin models, including the integration of immune components and endothelial cells, to accurately replicate the cutaneous microenvironment. Moreover, we discuss the potential of these models to identify novel therapeutic targets, enabling the design of personalized treatments aimed at restoring microbial balance, reinforcing the skin barrier, and modulating inflammation. As the field progresses, the incorporation of multi-omic approaches into skin-microbiome research will be pivotal in unraveling the complex interactions between host and microbiota, ultimately advancing therapeutic strategies for skin-related diseases. Full article

Utilizing straw feed is an effective strategy to optimize straw resource utilization by incorporating microbial degradation agents to expedite lignocellulose breakdown and enhance feed efficiency. Lignocellulose-degrading species and microbial communities are present in various Earth ecosystems, including the rumen of ruminants, insect digestive tracts, forest soil, and microbial populations in papermaking processes. The rumen of ruminants harbors a diverse range of microbial species, making it a promising source of lignocellulose-degrading microorganisms. Exploring alternative systems like insect intestines and forest soil is essential for future research. Current studies primarily rely on traditional microbial isolation techniques to identify lignocellulose-degrading strains, underscoring the necessity to transition to utilizing microbial culturomics and genome-editing technologies for discovering and manipulating cellulose-degrading microbes. This review provides an overview of lignocellulose-degrading microbial communities from diverse environments, encompassing bacterial and fungal populations. It also delves into the use of metagenomic, metatranscriptomic, and metaproteomic approaches to pinpoint highly efficient cellulase genes, along with the application of genome-editing tools for engineering lignocellulose-degrading microorganisms. The primary objective of this review is to offer insights for further exploration of potential lignocellulose-degrading microbial resources and high-performance cellulase genes to enhance roughage utilization in ruminant rumen ecosystems. Full article

Due to its high digestibility, rich nutrient profile, and potential probiotic content, goat milk is an essential nutritional resource, particularly for individuals with cow milk allergies. This review summarises the current state of microbial diversity in goat milk, emphasising the implications for quality, safety, and probiotic potential. This systematic review adhered to PRISMA guidelines, conducting a comprehensive literature search across PubMed, ScienceDirect, and Google Scholar using keywords related to microbial profiling in goat milk. The inclusion criteria targeted English-language studies from 2000 to 2025 that utilised high-throughput or next-generation sequencing methods. Out of 126 articles screened, 84 met the eligibility criteria. The extracted data focused on microbial diversity, profiling techniques, and their respective strengths and limitations in evaluating probiotic potential and spoilage risks. The review addresses the challenges linked to microbial spoilage and the composition and functional roles of microbial communities in goat milk. With species such as Bacillus and Pseudomonas playing crucial roles in fermentation and spoilage, key findings emphasise the prevalence of microbial phyla, including Proteobacteria, Firmicutes, and Actinobacteria in goat milk. The review also explores the probiotic potential of the goat milk microbiota, highlighting the health benefits associated with strains such as Lactobacillus and Bifidobacterium. Significant discoveries underline the necessity for advanced multi-omics techniques to thoroughly define microbial ecosystems and the substantial gaps in breed-specific microbiota research. Important findings illustrate the need for enhanced multi-omics techniques, given the challenges of host RNA and protein interference, low microbial biomass, and limited goat-specific reference databases, for optimising probiotic development, spoilage prevention strategies, and integrating metagenomics, metabolomics, metaproteomics, and metatranscriptomics to improve milk quality and safety as some of the future research objectives. This study emphasises the importance of understanding goat milk microbiology to advance dairy science and enhance human health. Full article

The human gastrointestinal tract harbors a vast array of microorganisms, which play essential roles in maintaining metabolic balance and immune function. While bacteria dominate the gut microbiome, fungi represent a much smaller, often overlooked fraction. Despite their relatively low abundance, fungi may significantly influence both health and disease. Advances in next-generation sequencing, metagenomics, metatranscriptomics, metaproteomics, metabolomics, and computational biology have provided novel opportunities to study the gut mycobiome, shedding light on its composition, functional genes, and metabolite interactions. Emerging evidence links fungal dysbiosis to various diseases, including inflammatory bowel disease, colorectal cancer, metabolic disorders, and neurological conditions. The gut mycobiome also presents a promising avenue for precision medicine, particularly in biomarker discovery, disease diagnostics, and targeted therapeutics. Nonetheless, significant challenges remain in effectively integrating gut mycobiome knowledge into clinical practice. This review examines gut fungal microbiota, highlighting analytical methods, associations with human diseases, and its potential role in precision medicine. It also discusses pathways for clinical translation, particularly in diagnosis and treatment, while addressing key barriers to implementation. Full article