Citrulline is the main precursor of a carcinogen compound ethyl carbamate (EC) in soy sauce. Accumulation of citrulline is mainly related to metabolism of arginine by Pediococcus acidilactici via the arginine deiminase (ADI) pathway during soy sauce fermentation. Elucidating the metabolic mechanism of ADI pathway that regulated by arc operon in P. acidilactici is essential for reduction of EC and its precursor in soy sauce. In this work, influences of pH, salt, ethanol and glucose on conversion of citrulline from arginine by P. acidilactici were investigated. Salt stress was the factor that most significantly influenced citrulline accumulation via the ADI pathway. Under salt stress, the ADI pathway in P. acidilactici exhibited a metabolic imbalance characterized by stronger inhibition of ornithine carbamoyltransferase (OTC) relative to arginine deiminase (ADI), directing substrate preferentially toward citrulline accumulation. Furthermore, the addition of ADI pathway activators (2, 4-dinitrophenol (DNP) and cyclic adenosine monophosphate (cAMP)) significantly increased arginine flux and suppressed intracellular citrulline accumulation; however, this inhibitory effect was abolished in the presence of 18% NaCl (w/v). These findings elucidate the mechanisms underlying the response of P. acidilactici to salt stress, revealing a dysregulation of the arc operon genes and key metabolic enzymes under high salt conditions. This insight provides a foundation for developing novel strategies to mitigate ethyl carbamate (EC) accumulation in soy sauce production.

Lactobacillus helveticus (L. helveticus) is a widely used probiotic valued for its applications in food fermentation. Dried bacterial inoculants, known for ease of transportation, stability, and flexible formulation, are increasingly favored. However, traditional spray drying, despite its continuous production mode, often results in significant loss of active components. In contrast, freeze-drying (FD) techniques, while preserving higher microbial viability, are limited by high energy costs and lengthy processing times. To address these challenges, this study employed an innovative electrostatic spray drying (ESD) technology for the preparation of L. helveticus inoculants. By optimizing microencapsulation techniques, heating temperature, and protective agents, a high-activity inoculant was developed using a novel spray-drying approach. The optimized inoculant achieved an initial survival rate of 82%, a production yield of 69.36%, residual moisture of 3.50%, water activity (aw) of 0.08, cell membrane integrity of 93.50%, and energy consumption of 7.5 kWh/L. After 3 months of storage at 4°C, survival rate remained at 79% with aw increasing to 0.22—well below the microbial growth threshold (aw 0.60)—and retained a smooth, crack-free microstructure. Fermentation performance analysis revealed that ESD-prepared inoculants outperformed freeze-dried inoculants. Although ESD extended the lag phase by 6 h compared to traditional liquid seed cultures, its fermentation efficiency remained comparable. This technology provides an efficient and standardized solution for large-scale production of L. helveticus in food fermentation, offering a promising alternative to conventional methods.

The anti-hyperuricemic, nephroprotective, and gut microbiota modulation effects of mixed carnosine and anserine (MAC) with the fixed ratio of 10:1 were evaluated in the hyperuricemic model mouse. The low- and high-dose (1.6 and 8.3 mg/kg BW) of MAC could significantly reduce serum uric acid (UA) level by 58% and 56%, close to the normal level, respectively. The MAC significantly inhibited xanthine oxidase activity in both hepatic tissue and serum. Furthermore, MAC treatment ameliorated the renal injury indicated by the normalized serum creatinine and urea nitrogen levels, recovered histopathological characteristics, mitigated oxidative stress, and suppressed proinflammatory cytokines. Notably, MAC could accelerate UA excretion by markedly downregulating the expression of the URAT1 and GLUT9, while upregulating the transporter OAT1. Gut microbiota analysis demonstrated that MAC restored intestinal microbial diversity in hyperuricemic mice and reduced the abundance of genera linked to hyperuricemia and inflammation. All the results proved MAC as potential UA-lowering treatment.

Dye-decolorizing peroxidases (DyPs) are heme-dependent oxidoreductases with promising applications in mycotoxin detoxification by acting on the oxidizable conjugated structures of aflatoxin B₁ (AFB₁). The heme prosthetic group is critical for the catalytic process by mediating electron transfer and intermediate formation that govern overall enzymatic activity. However, intracellular heme biosynthesis in microbial hosts is tightly regulated, frequently leading to suboptimal catalytic performance of recombinant DyPs, thus highlighting the necessity to improve cofactor availability. To establish an efficient heme-based expression system for DyPs, we developed a heme cofactor supplementation system in Escherichia coli by introducing a heterologous C4 pathway harboring 5-aminolevulinic acid synthase from Caulobacter segnis together with the outer membrane heme transporter ChuA. Furthermore, intracellular heme supply was optimized by the integrated overexpression of the C4 and C5 pathways as well as the hemEFGH gene cluster, which significantly enhanced RhDypB-R80 activity. The engineered cells exhibited substantially higher heme content and RhDypB-R80 activity compared to those produced by conventional cultivation methods supplemented with exogenous 5-ALA. Notably, heme-based RhDypB-R80 achieved 91.03% AFB₁ degradation within 24 h, demonstrating its potential for enzymatic detoxification applications. This engineered heme-producing host provides a practical platform for enhancing hemoprotein activity and supports the development of enzymatic strategies for mycotoxin control in food systems.

Enzyme immobilization serves as a pivotal strategy for enhancing enzymatic stability, reusability, and operational performance, demonstrating significant potential across diverse fields including biocatalysis, biosensing, food processing, environmental remediation, and biomedicine. This review systematically examines recent advances in enzyme immobilization, with a focus on advanced carrier platforms such as covalent organic frameworks (COFs), metal-organic frameworks (MOFs), natural and synthetic polymers, silica materials, magnetic materials, carbon materials, and corresponding composite materials. These materials provide robust support for constructing high-performance and multifunctional immobilized enzyme systems, as their tunable porosity, rich surface chemistry, excellent biocompatibility, and unique physicochemical properties. The review critically analyzes how various immobilization strategies, such as covalent bonding, physical adsorption, coprecipitation, and encapsulation, regulate enzymatic activity, stability, and selectivity. Moreover, the characteristics of these enzyme immobilization systems such as loading capacity, operational conditions and recyclability are summarized. It highlights the remarkable performance of these systems in applications spanning chiral synthesis, sensitive biosensing, pollutant degradation, drug delivery, food and beverage processing and continuous-flow bioprocessing. Finally, the review summarizes the current research landscape and highlights the most promising innovative prospects in this rapidly evolving field of enzyme immobilization.

This study rigorously assessed the impact of folic acid-fortified whole wheat products on mitigating mild liver injury caused by prolonged excessive folic acid consumption, improving folic acid bioavailability, and enhancing spatial cognitive functions in mice over a 48-day intervention period. In the water maze evaluation of the medium-dose folic acid-fortified group (M-B + FA), this group demonstrated the smallest escape latency (5.20 ± 1.49 s) and the longest length in the novel object recognition test (24.37 ± 17.21 s), signifying enhanced spatial memory and recognition capabilities. The whole wheat matrix markedly elevated folic acid concentrations in plasma (75.25 vs. 57.09 μg/mL) and cerebral tissue. The findings on serum antioxidant capacity indicated that folate-fortified whole wheat products markedly elevated superoxide dismutase (SOD) levels and decreased malondialdehyde (MDA). The liver function indicators indicated that the Folic acid group (FA) reduced aspartate aminotransferase (AST) and aminotransferase (ALT) levels, with minor vacuolation noted in the liver, which was mitigated by the consumption of whole wheat products. Histological evaluation revealed that folate supplementation resulted in a heightened quantity of Nissl bodies in the CA3 area of the hippocampus. Furthermore, metabolomic analysis demonstrated that constituents in whole grains, via synergistic interactions, stimulated pathways associated with vitamin digestion and absorption, cofactor biosynthesis, and neuroactive ligand-receptor interactions, consequently elevating the levels of neuroprotective metabolites such as tryptophan and bile acids. The whole grain diet actively enhances cognitive performance through synergistic interaction with folic acid by boosting its absorption, elevating systemic antioxidant levels and metabolic health, and regulating essential metabolic pathways associated with cognition.

Due to the increasing world population, traditional commercial sources of protein will soon be insufficient to meet global nutritional demands. To address this and other anthropological effects, including carbon emissions, land and freshwater usage and overfishing associated with traditional agriculture, aquaculture and wild-catch fisheries, cultivated meats have been proposed as a novel protein source, rather than animal slaughter. Currently, despite significant investment, the cultivated seafood industry is floundering to scale production. This is due to technological failures to adapt cells to suspension for integration into bioreactors, lack of appropriate scaffold materials and scalable techniques to support eventual product formation. To overcome these issues, it is crucial for the industry to choose methodologies and materials that are tailored to species-specific cells to compensate for differences in the sensitivity and behaviour of cells within different bioreactor systems. Moreover, for regulatory approval and consumer acceptance, appropriate materials and edibility are required for selecting scaffolds. Ultimately, the cultivated seafood industry must address the current limitations in production efficiency to become a viable alternative food source. To do so, the industry should focus on using the recent advancements from both industry and academia including methods and technologies in tissue engineering and the adaption of cells to suspension. This review focuses on cultivated seafood as an alternative to aquaculture and wild-catch fisheries and discusses the bottlenecks and potential solutions for the industry to upscale production.

Vicatia thibetica de Boiss (V. thibetica) is a medicinal and edible plant traditionally used by the Bai ethnic community in Yunnan, China. Previous studies have reported that extracts derived from its roots have anti-aging effects, although the active components remain to be further investigated. Our preliminary findings revealed that the V. thibetica crude polysaccharide (VTCP) extends the lifespan of nematodes. In the present study, we systematically investigated the anti-aging effects of VTCP in both d-galactose-induced and naturally aged mouse models with particular emphasis on the heart and kidneys as primary organs of interest. The results demonstrated that VTCP significantly reduced the expression of senescent biomarkers and alleviated fibrosis levels in both the heart and kidney. In the d-galactose-induced aging model, VTCP exhibited potent antioxidant and anti-inflammatory capacities, effectively lowering oxidative stress markers and the senescence-associated secretory phenotypes (SASPs) in cardiorenal tissues. More importantly, in naturally aged mice, VTCP remarkably reshaped the gut microbiota composition, characterized by a significant enrichment of beneficial bacteria with the short-chain fatty acids producing potential and a suppression of inflammation-associated genera. Based on these findings, we propose that the core mechanism of VTCP likely involves targeted modulation of the gut microbiota to promote the production of beneficial metabolites, which subsequently suppresses systemic oxidative stress and inflammation, ultimately leading to synergistic protection of cardiorenal tissues. Collectively, our results highlight the promising potential of VTCP in counteracting cardiorenal aging and implicate the modulation of the gut-heart/kidney axis as an underlying mechanism.

Oligosaccharides are increasingly valued in research and industry due to their structural diversity and health benefits. However, their efficient separation and purification pose significant challenges, primarily due to structural complexity and the limitations of traditional chromatographic methods. Membrane separation technology has emerged as a promising green and efficient alternative. By leveraging the selective permeability of membrane materials, this technology facilitates the separation, purification, and concentration of oligosaccharides with advantages such as lower energy consumption, operational simplicity, and high efficiency with oligosaccharide recovery often exceeding 70% and product purities frequently above 60% in integrated membrane processes. This systematic review discusses the application of membrane separation technology in oligosaccharide processing, focusing on four key techniques: microfiltration (MF), ultrafiltration (UF), nanofiltration (NF), and reverse osmosis (RO). It further delves into process optimization strategies, including membrane material selection, adjustment of operating parameters, and integrated system designs to enhance separation efficiency and product purity. Furthermore, it explores the future trends in addressing current challenges. By consolidating recent advancements and practical applications, this work provides valuable insights for researchers and industry practitioners, promoting the adoption of membrane separation technology for efficient and scalable oligosaccharide production.

Plant-based milks are beverages obtained through the aqueous extraction of plants, designed to replicate the appearance and texture of milk. These products have garnered significant attention due to their lower environmental impact, addressing concerns such as greenhouse gas emissions, water consumption, and land degradation. Plant-based milks align with the United Nations Sustainable Development Goals (SDGs), particularly SDG 12 (Responsible Consumption and Production) and SDG 13 (Climate Action), by offering sustainable alternatives that help reduce the environmental footprint of the food system. This review explores the production, fermentation, and potential applications of plant-based milks. It highlights the use of various plant species in the development of these products, detailing the fermentation process as a key innovation. Parameters such as microbial strains, temperature, and time are examined to demonstrate their impact on microbial kinetics, chemical composition, and quality. Additionally, the review discusses the challenges in improving physical and sensory properties and evaluates the potential of fermented plant-based milks as carriers for probiotics. Fermentation enhances the shelf life, chemical complexity, and health benefits of plant-based milks, fostering the production of bioactive, nutrient-rich beverages. Despite advances, physical and sensory challenges persist, necessitating further research. Fermented plant-based milks have shown promise as probiotic carriers, supported by in vitro and in vivo studies, though clinical trials are needed to fully validate their functionality. This rapidly growing field addresses consumer demand for sustainable and innovative products, contributing to global efforts to mitigate environmental impacts and promote sustainable food systems.