Search

Lactic acid has gained considerable attention globally due to its multi-purpose application. Commercial lactic acid production uses the fungal species Rhizopus oryzae, which produces other organic acids. A crucial point of effective fungal organic acid production is matching the fungal strains' requirements, where the carbon source plays a major role. The highest production rate is achieved when glucose is used as a carbon source. Alternatively, we can apply carbon-rich agricultural residues as carbon sources. Using agricultural waste for lactic acid production provides a sustainable and cost-effective feedstock but also helps to reduce greenhouse gas emissions by diverting waste from landfills and decreasing reliance on fossil fuels. Moreover, polylactic acid (PLA) produced from lactic acid monomers can occur in numerous agricultural applications. We should delve deeper into sustainable methods of using carbon residues to recycle waste, foster the circular economy, and advance sustainable agriculture. Therefore, there is a need for further research on the commercial use of agricultural and food industry wastes for lactic acid production. [ABSTRACT FROM AUTHOR]

Lactic acid has gained considerable attention globally due to its multi-purpose application. Commercial lactic acid production uses the fungal species Rhizopus oryzae, which produces other organic acids. A crucial point of effective fungal organic acid production is matching the fungal strains' requirements, where the carbon source plays a major role. The highest production rate is achieved when glucose is used as a carbon source. Alternatively, we can apply carbon-rich agricultural residues as carbon sources. Using agricultural waste for lactic acid production provides a sustainable and cost-effective feedstock but also helps to reduce greenhouse gas emissions by diverting waste from landfills and decreasing reliance on fossil fuels. Moreover, polylactic acid (PLA) produced from lactic acid monomers can occur in numerous agricultural applications. We should delve deeper into sustainable methods of using carbon residues to recycle waste, foster the circular economy, and advance sustainable agriculture. Therefore, there is a need for further research on the commercial use of agricultural and food industry wastes for lactic acid production.

The toll-like receptor 4 (TLR4) signaling pathway constitutes an intricate network of protein interactions primarily involved in inflammation and cancer. This pathway triggers intracellular signaling cascades, modulating transcription factors that regulate gene expression related to immunity and malignancy. Previous studies showed that colon cancer patients with low TLR4 expression exhibit extended survival times and the TLR4 signaling pathway holds a significant role in CRC pathogenesis. In recent years, traditional Chinese medicines (TCMs) have garnered substantial attention as an alternative therapeutic modality for CRC, primarily due to their multifaceted composition and ability to target multiple pathways. Emerging evidence indicates that specific TCM products, such as andrographolide, rosmarinic acid, baicalin, etc., have the potential to impede CRC development through the TLR4 signaling pathway. Here, we review the role and biochemical processes of the TLR4 signaling pathway in CRC, and natural products from TCMs affecting the TLR4 pathway. This review sheds light on potential treatment strategies utilizing natural TLR4 inhibitors for CRC, which contributes to the advancement of research and accelerates their clinical integration into CRC treatment. [ABSTRACT FROM AUTHOR]

Drought stress is an annual global phenomenon that has devastating effects on crop production, so numerous studies have been conducted to improve crop drought resistance. Plant-associated microbiota play a crucial role in crop health and growth; however, we have a limited understanding of the key processes involved in microbiome-induced crop adaptation to drought stress. In this review, we summarize the adverse effects of drought stress on crop growth in terms of germination, photosynthesis, nutrient uptake, biomass, and yield, with a focus on the response of soil microbial communities to drought stress and plant-microbe interactions under drought stress. Moreover, we review the morpho-physiological, biochemical, and molecular mechanisms underlying the mitigation effect of microbes on crop drought stress. Finally, we highlight future research directions, including the characterization of specific rhizosphere microbiome species with corresponding root exudates and the efficiency of rhizobacteria inoculants under drought conditions. Such research will advance our understanding of the complex interactions between crops and microbes and improve crop resistance to drought stress through the application of beneficial drought-adaptive microbes. [ABSTRACT FROM AUTHOR]

The toll-like receptor 4 (TLR4) signaling pathway constitutes an intricate network of protein interactions primarily involved in inflammation and cancer. This pathway triggers intracellular signaling cascades, modulating transcription factors that regulate gene expression related to immunity and malignancy. Previous studies showed that colon cancer patients with low TLR4 expression exhibit extended survival times and the TLR4 signaling pathway holds a significant role in CRC pathogenesis. In recent years, traditional Chinese medicines (TCMs) have garnered substantial attention as an alternative therapeutic modality for CRC, primarily due to their multifaceted composition and ability to target multiple pathways. Emerging evidence indicates that specific TCM products, such as andrographolide, rosmarinic acid, baicalin, etc., have the potential to impede CRC development through the TLR4 signaling pathway. Here, we review the role and biochemical processes of the TLR4 signaling pathway in CRC, and natural products from TCMs affecting the TLR4 pathway. This review sheds light on potential treatment strategies utilizing natural TLR4 inhibitors for CRC, which contributes to the advancement of research and accelerates their clinical integration into CRC treatment.

Abstract Background The Wnt signaling pathway is a complex network of protein interactions that functions most commonly in embryonic development and cancer, but is also involved in normal physiological processes in adults. The canonical Wnt signaling pathway regulates cell pluripotency and determines the differentiation fate of cells during development. The canonical Wnt signaling pathway (also known as the Wnt/β-catenin signaling pathway) is a recognized driver of colon cancer and one of the most representative signaling pathways. As a functional effector molecule of Wnt signaling, the modification and degradation of β-catenin are key events in the Wnt signaling pathway and the development and progression of colon cancer. Therefore, the Wnt signaling pathway plays an important role in the pathogenesis of diseases, especially the pathogenesis of colorectal cancer (CRC). Objective Inhibit the Wnt signaling pathway to explore the therapeutic targets of colorectal cancer. Methods Based on studying the Wnt pathway, master the biochemical processes related to the Wnt pathway, and analyze the relevant targets when drugs or inhibitors act on the Wnt pathway, to clarify the medication ideas of drugs or inhibitors for the treatment of diseases, especially colorectal cancer. Results Wnt signaling pathways include: Wnt/β-catenin or canonical Wnt signaling pathway, planar cell polarity (Wnt-PCP) pathway and Wnt-Ca2+ signaling pathway. The Wnt signaling pathway is closely related to cancer cell proliferation, stemness, apoptosis, autophagy, metabolism, inflammation and immunization, microenvironment, resistance, ion channel, heterogeneity, EMT/migration/invasion/metastasis. Drugs/phytochemicals and molecular preparations for the Wnt pathway of CRC treatment have now been developed. Wnt inhibitors are also commonly used clinically for the treatment of CRC. Conclusion The development of drugs/phytochemicals and molecular inhibitors targeting the Wnt pathway can effectively treat colorectal cancer clinically.

Microalgae are a vital resource for the coming years to address the concern of decrease in oil reserves and the negative impacts of fossil fuels on the environment. Their utilization is crucial for a wide range of industrial applications. Depending on the strain, microalgae contain a variety of chemical components and can be treated biochemically or thermochemically. This review thus focuses on the biochemical mechanisms that are used to convert algal biomass into sustainable fuel, including the challenges and potential of those processes. Microalgae have been shown to be a viable third-generation alternative to conventional biofuel feedstocks. The optimum production of biofuel depends on the proper selection of microalgae species based on their lipid, carbohydrate, and protein content in order to produce high-quality, sustainable biofuel. Nannochloropsis gaditana can contribute to a maximum biodiesel yield of 96.47%, whereas Nannochloropsis oculata can produce the least (25%) through the biochemical process of transesterification. Higher yields of microalgae-derived gaseous, solid and liquid fuels can be achieved by pre-treating microalgal biomass and then employing bioconversion processes such as photo-fermentation and hydrothermal carbonization.

Kang Chen,1 Song Zhang,2 Jinghua Jiao,3 Shan Zhao4 1Department of Ophthalmology, The First Hospital of China Medical University, Shenyang, Liaoning, People’s Republic of China; 2China Medical University, Shenyang, Liaoning, People’s Republic of China; 3Department of Anesthesiology, Central Hospital, Shenyang Medical College, Shenyang, Liaoning, People’s Republic of China; 4Department of Rheumatic Immunology, The First Hospital of China Medical University, Shenyang, Liaoning, People’s Republic of ChinaCorrespondence: Jinghua Jiao; Shan Zhao Email jean0905@163.com; zhaoshan1109@163.comAbstract: Lung cancer is characterized by high morbidity and mortality rates, and its occurrence is associated with many types of cell death. As a new form of regulated cell death, ferroptosis is an iron- dependent pattern of cell death and characterized by lethal accumulation of lipid-based reactive oxygen species (ROS), which is different from apoptosis, necrosis and autophagy at both the morphological and biochemical levels. It plays an important role in the development of lung cancer and induction of ferroptosis in lung cancer cells has become a new strategy for anti- lung cancer treatment. However, a few reviews summarized ferroptosis and its role in lung cancer has not been elucidated, and the precise mechanism of ferroptosis modeling lung cancer has not yet been revealed till date. Herein, we review the latest literature on the process of ferroptosis regarding lung cancer, including basic molecular or biology mechanistic studies both in vivo and in vitro, as well as human studies with a more translational or clinical approach. This review provides a practical, concise and updated outline on the mechanisms and therapeutic strategies in lung cancer with ferroptosis alterations. Looking ahead, further studies are required to uncover the possible modulatory relationship between ferroptosis and lung cancer.Keywords: programmed cell death, ferroptosis, biochemical process, lung cancer

Sludge accumulation in anaerobic lagoons is one of the major issues determining long-term operating costs. However, very little mechanistic analysis has been done on long-term sludge behaviour. A coupled hydrodynamic-biochemical model was developed using computational fluid dynamics (CFD), and results from this applied to a compartmental based model (CBM) for long-term simulation. The CFD model incorporates a mixture method for the spatial-and temporal evolution of fluid and solids with a non-Newtonian rheology. CFD was used to evaluate short term hydrodynamics, and a common CBM used to understand the fluid movement and sludge behaviour of full-scale anaerobic lagoons (with varying depths, sidewall slopes, and loading rates), operating in commercial piggeries located in Southern Queensland and Southern New South Wales, Australia. The results found that the lagoons had varying hydrodynamics, and sludge accumulates rapidly in sloped sidewall lagoons, forming a variable depth bed which occupied a substantial fraction of the lagoons. Shallow lagoons were dominated by significant surface recirculation dynamics, and were susceptible to solids accumulation, while deep lagoons allowed the formation of a well developed settled fraction. Predicted lagoon lifetimes varied substantially, but predicted long-term accumulation rates were approximately double that observed, due to long-term degradation of slowly degradable material. • A coupled CFD-Process Model developed for hydrodynamics and sludge accumulation. • The model revealed flow pattern and long-term sludge behaviour of full-scale WSLs. • The model predictions well complemented the experimental measurements. • The guideline provided for optimising lagoons design for desired throughputs. [ABSTRACT FROM AUTHOR]

Background: The Wnt signaling pathway is a complex network of protein interactions that functions most commonly in embryonic development and cancer, but is also involved in normal physiological processes in adults. The canonical Wnt signaling pathway regulates cell pluripotency and determines the differentiation fate of cells during development. The canonical Wnt signaling pathway (also known as the Wnt/β-catenin signaling pathway) is a recognized driver of colon cancer and one of the most representative signaling pathways. As a functional effector molecule of Wnt signaling, the modification and degradation of β-catenin are key events in the Wnt signaling pathway and the development and progression of colon cancer. Therefore, the Wnt signaling pathway plays an important role in the pathogenesis of diseases, especially the pathogenesis of colorectal cancer (CRC). Objective: Inhibit the Wnt signaling pathway to explore the therapeutic targets of colorectal cancer. Methods: Based on studying the Wnt pathway, master the biochemical processes related to the Wnt pathway, and analyze the relevant targets when drugs or inhibitors act on the Wnt pathway, to clarify the medication ideas of drugs or inhibitors for the treatment of diseases, especially colorectal cancer. Results: Wnt signaling pathways include: Wnt/β-catenin or canonical Wnt signaling pathway, planar cell polarity (Wnt-PCP) pathway and Wnt-Ca2+ signaling pathway. The Wnt signaling pathway is closely related to cancer cell proliferation, stemness, apoptosis, autophagy, metabolism, inflammation and immunization, microenvironment, resistance, ion channel, heterogeneity, EMT/migration/invasion/metastasis. Drugs/phytochemicals and molecular preparations for the Wnt pathway of CRC treatment have now been developed. Wnt inhibitors are also commonly used clinically for the treatment of CRC. Conclusion: The development of drugs/phytochemicals and molecular inhibitors targeting the Wnt pathway can effectively treat colorectal cancer clinically. [ABSTRACT FROM AUTHOR]

The global energy needs are increasing day by day to cater to the needs of the growing population. In spite of being the major energy source, conventional fossil fuels have strong negative impact on the environment. To mitigate this, non-conventional energy sources like biomethanol having many relative advantages can replace them. Employing various thermochemical and biochemical process, biomethanol can be produced from waste biomass like agrowastes, forest litter, and urban organic wastes, after a judicious pretreatment to ensure higher yield. Pretreatment especially become a predisposing factor when complex lignocellulosic biomass is involved as a substrate. Methanol is produced via synthesis gas (syngas) in thermochemical process, and through anaerobic digestion in the presence of methanotrophic bacteria in biochemical process. During the production of methanol from syngas involving the former process, few high pressure (ZnO/Cr2O3) and low-pressure (Cu/ZnO and Cu/ZnO/Al2O3) catalysts are employed. Selecting the most potent methanotrophs is critical in the latter process. In this process, a critical enzyme in methanol production is methane monooxygenase (MMO) which helps in converting methane to methanol. A variety of soil methanotrophic bacteria have been assessed by using high throughput sequencing of pmoA gene. To make the process cost-effective, raw biogas is used as a substrate rather than natural gas to produce methanol. Possibilities of utilising lignocellulosic biomass through biochemical route to produce methanol seem to be a more promising approach. [ABSTRACT FROM AUTHOR]

The anaerobic digestion technology has been established decades ago but still the commercial biogas industries are facing some challenges like process imbalance and failure of the digester, which has to be addressed by monitoring and identifying early warning indicators essential in the operation and maintenance. The aim of present study is to evaluate the early warning indicators during start-up: optimization, full load, and gradual overload phases of an 18-m3 commercial food waste biogas plant. The plant was operated by applying the optimal operation limits of FOS/TAC, TVFA/TA, IA/PA (Reply's), and BA/TA ratios in the optimization of the AD process. The plant performance was studied in terms of methane yield, reduction in OLR, HRT, VS, and COD. The plant was operated for a span of 12 months at different OLRs ranging from 0.32 kg COD/m3/day (start-up) to 2.03 kg COD/m3/day (overload) in a mesophilic condition. Results showed that performance of AD at full load, in terms of biogas yield (m3/day) and methane yield (m3 CH4/(kg VS reduced) and m3 CH4/(kg COD reduced)), was found to be average of 32.65, 0.55, and 0.57, respectively. The process imbalance was observed from day 114 to day 119 of optimization period. The values of FOS/TAC, IA/PA, and TVFA/TA ratios were detected to be slightly more than the regular values, whereas bicarbonate alkalinity (BA) was abruptly reduced to 0.37 (kg CaCO3/m3). On the other hand, intermediate alkalinity (IA) value has been increased to 3.2 (kg CaCO3/m3), which was defined as limit for process instability. The gradual overload (0.6% more VS addition for 28 days) has no significant affect on the AD process and biogas production, but methane yield was slightly decreased. [ABSTRACT FROM AUTHOR]

Ferroptosis is a newly discovered mechanism of cell death characterized by accumulation of iron dependent lipid peroxidation and reactive oxygen species, which is different from apoptosis, necrosis and autophagy in morphology and biochemical findings. It is involved in the occurrence and development of lung cancer, so inducing ferroptosis seems to become a new strategy for anti-lung cancer therapy. This review includes the followings: mechanism of ferroptosis from the aspects of iron metabolism, amino acid and glutathione metabolism and lipid metabolism; its effects on lung cancer occurrence and progression, chemotherapy resistance, radiation resistance and immunotherapy respectively, in order to update information on ferroptosis and thus provides a new direction and new strategy for lung cancer treatment.

Purpose: Organic amendments greatly influence soil functions and biochemical processes that are driven by soil active microbiota. Yet, how organic fertilization impacts transcriptionally active microbes and impacts soil ecosystem functions is still largely unknown. In this study, the variations of RNA-based soil active bacteria and multifunctionality in response to long-term organic fertilization were investigated. Methods: We selected an experimental observation station with the application of organic fertilizers for 30 years. High-throughput sequencing and qPCR-based SmartChip assays were employed to explore the abundance, diversity, composition, and function of soil active microorganisms under different application regimes. Results: Organic fertilization increased soil transcriptionally active bacterial abundances significantly, and changed their compositions compared to control treatment. Of the differentially active bacteria, Bacillus was the most abundant genus which was increased by organic fertilization dramatically. Unexpectedly, however, the impacts of organic fertilization on the active communities did not demonstrate a dose-dependent response at the RNA level. SmartChip analysis further indicated that organic fertilization significantly increased the abundances of 25 functional genes associated with energy metabolism, organic matter degradation, denitrification, phosphorus solubilization, sulfur oxidation, and sulfate reduction processes. In addition, soil multifunctionality was also promoted by organic fertilization, and such promotion had a strong relationship with the differentially active bacteria, suggesting that the variations of soil ecosystem functions could be mainly regulated by differentially active bacterial communities. Conclusion: Organic fertilization impacted the abundances and compositions of soil active bacteria, and increased soil multifunctionality. Understanding dynamic changes in soil active microbes is therefore crucial for managing application regimes of organic fertilizers in agricultural practices and evaluating element biogeochemical cycling in soil. [ABSTRACT FROM AUTHOR]

Passion fruit (Passiflora edulis Sims) is a tropical fruit that shows an increasing demand from global fresh fruit industries. The fruit is known to have climacteric properties that allow it to achieve ripening during distribution and transportation. However, the metabolic alterations that occur during ripening are poorly understood. Thus, this study was conducted to analyze the metabolites in passion fruit juice at different harvesting stages (35, 42, 49, 56 and 63 days after anthesis), employing 1H NMR spectroscopy combined with chemometric analysis. A total of 30 metabolites were successfully identified using 1H NMR, the majority of which belong to primary metabolites, consisting of 14 amino acids, 7 sugars and 6 organic acids. Nevertheless, only three secondary metabolites were detected. Analyzing the metabolites using multivariate analysis reveals 13 of the metabolites were highly influential metabolites. These primary metabolites play essential roles in the maturation and ripening of fruit. Glycolysis, the tricarboxylic acid (TCA) cycle and the shikimate pathway were differentially regulated during maturation and ripening of passion fruit. Profiling the metabolome of fruit during maturation and ripening reveals that all metabolites are interconnected with each other to cause maturation and ripening in passion fruit. This work will enhance the knowledge of physiological changes in purple passion fruit during maturation and ripening.

Aquilaria malaccensis is a precious tropical plant reputed for its agarwood resin, which is the outcome of biochemical interaction of the plant with fungus. The present study was aimed at investigating the process of agarwood production by isolating endophytic fungus from juvenile plant, effect of stem pieces of juvenile plant on the growth of fungus, and antibacterial effect of isolated fungus and leaves. Four endophytic fungi (Alternaria, Curvularia, Rhizopus and Sterilia species) were isolated from juvenile (1-year-old) A. malaccensis plant and three fungal colonies (Penicillium, Fusarium and one putative Cladosporium species) were isolated from agarwood chips. The addition of stem pieces of juvenile A. malaccensis to the culture media (PDA) exhibited profuse and faster growth of six different fungi. The methanolic leaf extract (concentration 200 mL) showed clear inhibitory zone against Streptococcus pneumonia (15.66 mm), Escherichia coli (12.80 mm), and Shigella dysenteriae (13.66 mm). However, 100 µL of methanolic leaves extract showed inhibition of 12.66, 12.30 and 11.41 mm on S. pneumonia, E. coli and S. dysenteriae plates respectively. The study concludes that agarwood is the product of persistent interaction between endogenous fungi and the plant, which triggers at early stage of its growth and the resin production probably is visible at onset of maturity. [ABSTRACT FROM AUTHOR]

Morphological changes and biochemical process of ramie retting by RAMCD407 was investigated using microscopy techniques, including OM, PLM, FLSM and SEM coupled with enzyme activity analysis. Results showed that retting was completed within 56 h followed by 0.2% NaOH treatment. Residual gum content and breaking strength of the final fiber was 2.84% and 5.2 cN/dtex, respectively. This fulfills the requirement for ramie spinning. Retting included four main processes: water absorption and CCCO formation; cortex removal; removal of gum in the middle lamella; and removal of gum on the surface of fiber. The first two processes were completed under low enzyme activity condition, while the third process was related to higher pectinase enzyme activity and the last process was related to higher enzyme activities of pectinase and xylanase. In fact, only a small fraction of hemicellulose and pectin is required to be hydrolyzed to ensure fiber separation. In this process, pectinase played a key role supported by xylanase. Valuable retting residues, such as pectinase crystals, pectin blocks, xylan chips and microcrystalline cellulose was recovered from residual liquor by filtration and/or centrifugation. Reuse of these waste residues can help increase the overall economic benefit for ramie industries. The occurrence of spherical balls with double shell structure was found in retting process, and FLSM showed that the outer shell was composed of xylan-rich material. Compared to traditional chemical degumming methods, ramie retting by RAMCD407 is an eco-friendly degumming technique. [ABSTRACT FROM AUTHOR]

Information about the response of soil enzymes to field warming in permafrost regions is scarce, and the potential mechanisms by which warming combined with biotic and abiotic factors affect soil enzyme activities in alpine grasslands remain unclear. A 3-year in situ experiment with two warming levels (2.7 °C and 5.3 °C) was conducted in an alpine swamp meadow to investigate the effects of experimental warming on 5 soil enzyme activities involved in soil carbon and nitrogen cycling, and the relationships between soil enzyme activities and soil variables related to physicochemical properties and nutrient levels were examined. Results showed that warming had obvious positive effects on soil moisture (SM), soil organic carbon, total nitrogen, and NH 4 +-N contents, especially in the surface soil layer. Soil NO 3 --N tended to decrease under moderate warming, while it significantly increased under high warming. Meanwhile, during the entire growing season, warming strongly enhanced invertase and amylase activities by 39.2–49.0% and 109.9–191.0%, respectively. In contrast, urease activities were significantly decreased by 36.8–55.7% in warming plots and there were no significant differences in catalase or cellulase activities among treatments during the whole growing season, while catalase activities in warming plots were significantly decreased by 2.6–4.0% in June. These inconsistent responses of soil enzyme activities to experimental warming could be partially explained by warming-induced changes in soil variables. Redundancy analysis showed that soil NO 3 --N and SM were the most important factors, which explained 29.8% and 19.7% of the variations in soil enzyme activities, respectively, suggesting that the warming-induced increase in SM could weaken the soil aeration status and enhance enzyme and substrate diffusion, strongly affecting soil nitrification and microbial enzyme production. • Warming strongly influenced soil C and N stocks and availability at least in the surface soil layer in an alpine swamp meadow. • Soil enzyme activities showed inconsistent responses to experimental warming. • Soil NO 3 --N content and soil moisture (SM) were key factors controlling the variations in soil enzyme activities. [ABSTRACT FROM AUTHOR]

Biochemical processes present complex mechanisms and can be described by various computational models. Complex systems present a variety of problems, especially the loss of intuitive understanding. The present work uses fractional-order calculus to obtain mathematical models for erythritol and mannitol synthesis. The obtained models are useful for both prediction and process optimization. The models present the complex behavior of the process due to the fractional order, without losing the physical meaning of gain and time constants. To validate each obtained model, the simulation results were compared with experimental data. In order to highlight the advantages of fractional-order models, comparisons with the corresponding integer-order models are presented.

This study assessed the components of anaerobically digested sludge, activated sludge, and microbial and extracellular polymeric substance (EPS) enzymes to identify the mechanisms underlying nitrogen removal and soil regeneration. 16S rRNA gene amplicon-based sequencing was used to determine the microbial community composition and the related National Center for Biotechnology Information (NCBI) protein database was used to construct a conventional library from the observed community. EPS components were identified using gel-free proteomic (Liquid Chromatography with tandem mass spectrometry-LC/MS/MS) methods. Alginate-like EPS from aerobically activated sludge have strong potential for soil aggregation and water-holding capacity, whereas total EPS from anaerobic sludge have significant potential for ammonia removal under salt stress. Fourier transform infrared spectroscopy (FTIR) revealed that both EPS may contain proteins, carbohydrates, humic compounds, uronic acid, and DNA and determined the presence of O–H, N–H, C–N, C O, and C–H functional groups. These results demonstrate that the overall enzyme activity may be inactivated at 30 g L-1 of salinity. An annotation found in Kyoto Encyclopedia of Genes and Genomes (KEGG)- KEGG Automatic Annotation Server (KAAS) revealed that the top two metabolic activities in the EPS generated from the anaerobic sludge were methane and nitrogen metabolism. Therefore, we focused on the nitrogen metabolism reference map 00910. EPS from the anaerobically digested sludge exhibited nitrate reductase, nitrite reductase, and dehydrogenase activities. Assimilatory nitrate reduction, denitrification, nitrification, and anammox removed ammonia biochemically. The influence of microbial extracellular metabolites on water-holding capacity and soil aggregation was also investigated. The KAAS-KEGG annotation server was used to identify the main enzymes in the activated sludge-derived alginate-like extracellular EPS (ALE-EPS) samples. These include hydrolases, oxidoreductases, lyases, ligases, and transporters, which contribute to soil fertility and stability. This study improves our understanding of the overall microbial community structure and the associated biochemical processes, which are related to distinct functional genes or enzymes involved in nitrogen removal and soil aggregation. In contrast to conventional methods, microbial association with proteomics can be used to investigate ecological relationships, establishments, key player species, and microbial responses to environmental changes. Linking the metagenome to off-gel proteomics and bioinformatics solves the problem of analyzing metabolic pathways in complex environmental samples in a cost-effective manner. [Display omitted] • Sludge derived EPS enzymes were investigated to assess nitrogen removal and soil regeneration. • Gel-free proteomics and microbial compositions revealed EPS components and functions. • EPS from anaerobic sludge removed ammonia by nitrate, nitrite reductase, and dehydrogenase. • KAAS-KEGG identified soil-fertility-boosting enzymes in activated sludge-derived EPS. • Microbial proteome analyses ecological linkages, key-players, and their responses to environment. [ABSTRACT FROM AUTHOR]