Degradation Of Aflatoxin B1 Research Articles

Degradation of Aflatoxin B1 by the Armillariella tabescens-derived aldo-keto reductase AtAKR

Aflatoxin B1 (AFB1) is one of the most toxic mycotoxins discovered to date and is prevalent in various geographical regions. Armillaria tabescens has been substantiated to be capable of degrading AFB1, with the degradation process potentially attributable to aldo-keto reductases. Therefore, this study aimed at assessing the AFB1-degrading efficacy of the aldo-keto reductase AtAKR through its cloning and recombinant expression in A. tabescens. The entire AtAKR open reading frame spans a length of 954 bp, which corresponds to 317 encoded amino acids. The in vitro analysis of the interaction between AtAKR and AFB1 revealed that AtAKR exhibits direct degradation of AFB1 (100 ppb) in the presence of NADPH. The degradation rates observed were 24.3% and 34.06% within 12-h and 36-h periods, respectively. Liquid chromatography-mass spectrometry was employed for the identification of the degradation products, and the results indicated that AFB1 can undergo direct degradation to Aflatoxicol, which is considerably less toxic compared to AFB1. These findings present AtAKR as a promising candidate for a potential application in the degradation of AFB1 present in food and feed.

Bacillus licheniformis ameliorates Aflatoxin B1-induced testicular damage by improving the gut-metabolism-testis axis

Global aflatoxin B1 (AFB1) contamination is inevitable, and it can significantly damage testicular development. However, the current mechanism is confusing. Here, by integrating the transcriptome, microbiome, and serum metabolome, we comprehensively explain the impact of AFB1 on testis from the gut-metabolism-testis axis. Transcriptome analysis suggested that AFB1 exposure directly causes abnormalities in testicular inflammation-related signalling, such as tumor necrosis factor (TNF) pathway, and proliferation-related signalling pathways, such as phosphatidylinositide 3-kinases-protein kinase B (PI3K-AKT) pathway, which was verified by immunofluorescence. On the other hand, we found that upregulated inflammatory factors in the intestine after AFB1 exposure were associated with intestinal microbial dysbiosis, especially the enrichment of Bacilli, and enrichment analysis showed that this may be related to NLR family pyrin domain containing 3 (NLRP3)-mediated NOD-like receptor signalling. Also, AFB1 exposure caused blood metabolic disturbances, manifested as decreased hormone levels and increased oxidative stress. Significantly, B. licheniformis has remarkable AFB1 degradation efficiency (> 90%). B. licheniformis treatment is effective in attenuating gut-testis axis damage caused by AFB1 exposure through the above-mentioned signalling pathways. In conclusion, our findings indicate that AFB1 exposure disrupts testicular development through the gut-metabolism-testis axis, and B. licheniformis can effectively degrade AFB1.

Bacillus subtilis Simultaneously Detoxified Aflatoxin B1 and Zearalenone

The co-occurrence of aflatoxin B1 (AFB1) and zearalenone (ZEN) in grain-based food and animal feed poses significant health risks to humans and animals due to their potent mutagenic, cytotoxic, and carcinogenic properties. Conventional physical and chemical methods are insufficient for effectively detoxifying multiple mycotoxins present in food and feed. In this study, we evaluated the capability of Bacillus subtilis ZJ-2019-1 (B. subtilis ZJ-2019-1) to simultaneously degrade AFB1 and ZEN while optimizing reaction to enhance degradation efficiency. The localization of active ingredients from B. subtilis ZJ-2019-1 was determined using high-performance liquid chromatography (HPLC). Our findings demonstrated that B. subtilis ZJ-2019-1 eliminated 60.88% of AFB1 and 33.18% of ZEN within 72 h at a concentration of 10 mg/L at 37 °C (pH 7.0) and exerted greater activity under alkaline conditions. The autoclaved and boiled supernatants of B. subtilis ZJ-2019-1 exhibited significant enhancement in the degradation of AFB1 and ZEN, achieving degradation rates of 79.85% and 100%, respectively, at a concentration of 1 mg/L within 48 h at 37 °C. Moreover, the crude enzymes from B. subtilis ZJ-2019-1 showed maximum degradation rates for AFB1 (100%) and ZEN (94.29%) within 72 h at 70 °C. Additionally, divalent cations (such as Co2+, Fe2+, Mn2+, and Ni2+) significantly augmented the activity of crude enzymes from B. subtilis ZJ-2019-1 towards mycotoxin degradation. Furthermore, when applied to corn gluten meals, B. subtilis ZJ-2019-1 strain effectively detoxify 66.08% of AFB1 and 22.01% of ZEN, surpassing the efficacy of a commercial detoxification agent on the market (34.17% for AFB1 and 2.28% for ZEN). Collectively, these findings indicated that B. subtilis ZJ-2019-1 is a promising candidate for the simultaneous removal of multiple mycotoxins in food and feed, while addressing health concerns associated with harmful mycotoxins.

Characterization and mechanism of simultaneous degradation of aflatoxin B1 and zearalenone by an edible fungus of Agrocybe cylindracea GC-Ac2.

Contamination with multiple mycotoxins is a major issue for global food safety and trade. This study focused on the degradation of aflatoxin B1 (AFB1) and zearalenone (ZEN) by 8 types of edible fungi belonging to 6 species, inclulding Agaricus bisporus, Agrocybe cylindracea, Cyclocybe cylindracea, Cyclocybe aegerita, Hypsizygus marmoreus and Lentinula edodes. Among these fungi, Agrocybe cylindracea strain GC-Ac2 was shown to be the most efficient in the degradation of AFB1 and ZEN. Under optimal degradation conditions (pH 6.0 and 37.4°C for 37.9 h), the degradation rate of both AFB1 and ZEN reached over 96%. Through the analysis of functional detoxification components, it was found that the removal of AFB1 and ZEN was primarily degraded by the culture supernatant of the fungus. The culture supernatant exhibited a maximum manganese peroxidase (MnP) activity of 2.37 U/mL. Interestingly, Agrocybe cylindracea strain GC-Ac2 also showed the capability to degrade other mycotoxins in laboratory-scale mushroom substrates, including 15A-deoxynivalenol, fumonisin B1, B2, B3, T-2 toxin, ochratoxin A, and sterigmatocystin. The mechanism of degradation of these mycotoxins was speculated to be catalyzed by a complex enzyme system, which include MnP and other ligninolytic enzymes. It is worth noting that Agrocybe cylindracea can degrade multiple mycotoxins and produce MnP, which is a novel and significant discovery. These results suggest that this candidate strain and its enzyme system are expected to become valuable biomaterials for the simultaneous degradation of multiple mycotoxins.

Photocatalytic degradation of aflatoxin B1 by porous carbon nitrides under visible light

Photocatalytic degradation of aflatoxin B1 by porous carbon nitrides under visible light

Development of Plasma-Activated Water (PAW) System: Molecular Dynamics Simulation and Experimental Study on Almond Aflatoxins

Almonds play a significant role in Iran’s economy, and factors that threaten their market, such as aflatoxins, need careful consideration. Plasma-activated water (PAW) is a new method that has antioxidant activity and can eliminate toxins and microbial agents, making it a suitable solution for removing aflatoxins from almonds. In a study, the continuous PAW production system was used to control and remove almond aflatoxins, and molecular dynamics simulation was employed to evaluate the impact of PAW on aflatoxin B1. The study found that reducing the flow rate of plasma-treated water had the greatest effect on reducing aflatoxin concentration, followed by PAW application time, the dose of inoculated toxin, and the air/gas mixture ratio. The use of PAW reduced aflatoxin concentration in almonds by 12% to 56.8%. The simulation results suggested that PAW can affect the structure of aflatoxin B1, change and destroy its activity, and reduce its toxicity. Among the free radicals tested, NO3− was found to be the most effective in degrading aflatoxin B1. This study demonstrates the potential of PAW as a method to remove aflatoxins and enhance the safety of almonds.

Immobilization of recombinant Trametes versicolor aflatoxin B1-degrading enzyme (TV-AFB1D) with montmorillonite for absorption and in situ degradation of aflatoxin B1.

Aflatoxin B1 is a highly carcinogenic and teratogenic substance mainly produced by toxin-producing strains such as Aspergillus flavus and Aspergillus parasitic. The efficient decomposition of aflatoxin is an important means to reduce its harm to humans and livestock. In this study, Trametes versicolor aflatoxin B1-degrading enzyme (TV-AFB1D) was recombinantly expressed in Bacillus subtilis (B. subtilis) 168. MMT-CTAB-AFB1D complex was prepared by the immobilization of TV-AFB1D and montmorillonite (MMT) by cross-linking glutaraldehyde. The results indicated that TV-AFB1D could recombinantly express in engineered B. subtilis 168 with a size of approximately 77kDa. The immobilization efficiency of MMT-CTAB-AFB1D reached 98.63% when the concentration of glutaraldehyde was 5% (v/v). The relative activity of TV-AFB1D decreased to 72.36% after reusing for 10 times. The content of AFB1 in MMT-CTAB-AFB1D-AFB1 decreased to 1.1µg/g from the initial 5.6µg/g after incubation at 50°C for 6h. The amount of 80.4% AFB1 in the MMT-CTAB-AFB1D-AFB1 complex was degraded by in situ catalytic degradation. Thus, the strategy of combining adsorption and in situ degradation could effectively reduce the content of AFB1 residue in the MMT-CTAB-AFB1D complex.

Highly efficient g-C3N4 catalysts derived from various precursors for aflatoxin B1 degradation under visible light

A g-C3N4 catalyst derived from urea had excellent photocatalytic degradation efficiency (93.5%) for AFB1 within 30 min. The deactivation mechanism of AFB1 during photodegradation was also investigated.

Synthesis of visible light driven spherical Bi2MoO6 structure for aflatoxin B1 photodegradation

Aflatoxin B1 (AFB1) is a dangerous toxin found in food and feed, it is of great significance to find a photocatalyst for the efficient degradation of AFB1. Bismuth molybdate (Bi2MoO6) photocatalyst has advantages of good visible light response, low cost, and non-toxicity. In this study, the solvent composition polarity and different temperatures were discussed on the surface nano-sheets aggregation, crystal size, and crystal defect of Bi2MoO6. The photocatalytic performance of Bi2MoO6 was investigated towards AFB1 degradation under visible light. The results showed that sample BMO(3:7)-160 prepared under a volume ratio of 3:7 about ethylene glycol to ethanol and 160℃ was spherical structure composed of nanosheets, which had a good photocatalytic degradation efficiency and photocatalytic stability on AFB1. After 120 min, the degradation rate of AFB1 can reach 99.8 %, which was attributed to the best electron-hole separation efficiency of BMO(3:7)-160. In addition, h+ was identified as the primary active species. The degradation products of AFB1 were analyzed to deduce the pathway of AFB1 decomposition and the possible photocatalytic mechanism.

Mining Lactonase Gene from Aflatoxin B1-Degrading Strain Bacillus megaterium and Degrading Properties of the Recombinant Enzyme.

Mycotoxins are toxic secondary metabolites mainly produced by filamentous fungal species that commonly contaminate food and feed. Aflatoxin B1 (AFB1) is extremely toxic and seriously threatens the health of humans and animals. In this work, the Bacillus megaterium HNGD-A6 was obtained and showed a 94.66% removal ability of AFB1 by employing extracellular enzymes as the degrading active substance. The degradation products were P1 (AFD1, C16H14O5) and P2 (C14H16N2O2), and their toxicity was greatly reduced compared to that of AFB1. The AttM gene was mined by BlastP comparison and successfully expressed in Escherichia coli BL21. AttM could degrade 86.78% of AFB1 at pH 8.5 and 80 °C, as well as 81.32% of ochratoxin A and 67.82% of zearalenone. The ability of AttM to degrade a wide range of toxins and its resistance to high temperatures offer the possibility of its use in food or feed applications.

Recombinant Oxidase from Armillaria tabescens as a Potential Tool for Aflatoxin B1 Degradation in Contaminated Cereal Grain.

Forage grain contamination with aflatoxin B1 (AFB1) is a global problem, so its detoxification with the aim of providing feed safety and cost-efficiency is still a relevant issue. AFB1 degradation by microbial enzymes is considered to be a promising detoxification approach. In this study, we modified an previously developed Pichia pastoris GS115 expression system using a chimeric signal peptide to obtain a new recombinant producer of extracellular AFB1 oxidase (AFO) from Armillaria tabescens (the yield of 0.3 g/L), purified AFO, and selected optimal conditions for AFO-induced AFB1 removal from model solutions. After a 72 h exposure of the AFB1 solution to AFO at pH 6.0 and 30 °C, 80% of the AFB1 was degraded. Treatments with AFO also significantly reduced the AFB1 content in wheat and corn grain inoculated with Aspergillus flavus. In grain samples contaminated with several dozen micrograms of AFB1 per kg, a 48 h exposure to AFO resulted in at least double the reduction in grain contamination compared to the control, while the same treatment of more significantly (~mg/kg) AFB1-polluted samples reduced their contamination by ~40%. These findings prove the potential of the tested AFO for cereal grain decontamination and suggest that additional studies to stabilize AFO and improve its AFB1-degrading efficacy are required.

DypB peroxidase for aflatoxin removal: New insights into the toxin degradation process

Aflatoxin B1 (AFB1) is one of the most potent carcinogens and a widespread food and feed contaminant. As for other toxins, many efforts are devoted to find efficient and environmentally-friendly methods to degrade AFB1, such as enzymatic treatments, thus improving the safety of food and feed products. In this regard, the dye decolorizing peroxidase of type B (DypB) can efficiently degrade AFB1. The molecular mechanism, which is required to drive protein optimization in view of the usage of DypB as a mycotoxin reduction agent in large scale application, is unknown. Here, we focused on the role of four DypB residues in the degradation of AFB1 by alanine-scanning (residues 156, 215, 239 and 246), which were identified from biochemical assays to be kinetically relevant for the degradation. As a result of DypB degradation, AFB1 is converted into four products. Interestingly, the relative abundancy of these products depends on the replaced residues. Molecular dynamics simulations were used to investigate the role of these residues in the binding step between protein and manganese, a metal ion which is expected to be involved in the degradation process. We found that the size of the haem pocket as well as conformational changes in the protein structure could play a role in determining the kinetics of AFB1 removal and, consequently, guide the process towards specific degradation products.

Radio frequency roasting promotes the degradation of aflatoxin B1 and achieves better quality of peanuts (Arachis hypogaea L.)

Peanuts contaminated with aflatoxin B1 (AFB1) possess serious health risks. Radio frequency (RF) roasting was used for decontamination of AFB1 in peanuts. The effects of RF roasting on AFB1 degradation and peanuts quality were investigated under varied electrode gaps (90, 100, 110, and 120 mm), target temperatures (120, 130, 140, and 150 °C) and initial moisture contents (10%, 15%, 20%, and 25%). The results showed that corn grit as the assistant material was superior for improving the heating uniformity of peanuts. With increasing target temperature and moisture content, the degradation rate of AFB1 increased significantly from 20.05% to 75.33% and 35.52%–66.09%, respectively, while electrode gap had no significant effect on the degradation of AFB1. Compared with hot air roasting (140 °C), RF roasting increased the degradation rate of AFB1 by 18%, with lower acid value and less tissue damage. Fourier transform infrared spectrum and scanning electron micrographs revealed the development of lipid hydrolysis, Maillard reaction and protein denaturation in roasted peanuts. RF roasting under 110–120 mm electrode gap, 140 °C target temperature, and 15%–20% initial moisture content was optimal considering both AFB1 degradation and products quality. Overall, RF roasting provides a new insight for the degradation of AFB1 in foods and crops.

Activate hydrogen peroxide for facile and efficient removal of aflatoxin B1 by magnetic Pd-chitosan/rice husk-hercynite biocomposite and its impact on the quality of edible oil

Due to the high heat and chemical stability of aflatoxin B1 (AFB1) with significant impacts on humans/animals and thus it needs to develop a practical and efficient approach for its removal. Herein, we fabricated a magnetic Pd-chitosan/glutaraldehyde/rice husk/hercynite (Pd@CRH-x) composite for efficient detoxification of AFB1. The Pd@CRH-x was obtained by a simple wet-impregnation procedure of CRH complexes followed by pyrolysis. The results confirmed that the unique structure of Pd@CRH-400 effectively improves dispersity, and mass transfer subsequently enhancing removal efficiency in batch conditions. Results indicate 94.30 % of AFB1 was efficiently degraded by 0.1 mg mL−1 Pd@CRH-400 with 4.0 mM H2O2 at wide pH ranges (3.0–10) at 60 min with a decomposition rate constant of 0.0467 min−1. Besides, by comparing the quality factors of edible oil (i.e., acid value, peroxide value, iodine value, moisture, volatile matters, anisidine value, and fatty acid composition), it was confirmed that there was no obvious influence on the physicochemical indicators of edible oil after removal/storage process. Subsequently, the systematic kinetic study and AFB1 degradation mechanism were presented. This study provides a new strategy for the efficient construction of controllable and dispersed Pd-based catalysts using CRH-x as a spatial support for alleviating the risk of toxic pollutants.

Heat stability of Trichoderma asperelloides SKRU-01 culture filtrates: Potential applications for controlling fungal spoilage and AFB1 production in peanuts

This study aimed to examine the heat stability of culture filtrates of Trichoderma asperelloides SKRU-01 (culture filtrates SKRU-01) over a temperatures range (40–121 °C) and the effects on the antifungal activity against two aflatoxin-producing strains (Aspergillus parasiticus TISTR 3276 and A. flavus PSRDC-4), aflatoxin B1 (AFB1) degradation, and the role in mycotoxin control in peanuts. The impact of SKRU-01 culture age (2–12 day-old) on both pathogenic strains revealed that the culture age of 6–12 day-old cultures exhibited no significant difference (p > 0.05) of growth inhibition for strain TISTR 3276 (81.89–82.28 %) and 4–12 day-old cultures for strain PSRDC-4 (74.87–79.06 %). The heat-treated temperatures from 40 °C to 121 °C caused no significant (p > 0.05) reduction of mycelial growth for strain TISTR 3276 (82.61 % to 79.13 %) but significant (p < 0.05) deduction for strain PSRDC-4 (75.15 % to 59.17 %). Heat treatment of the culture filtrates SKRU-01 at 60–121 °C caused the reduction on spore germination inhibition (from about 68 % to 58.16 % for strain TISTR 3276 and 51.11 % for strain PSRDC-4). These results indicate that strain TISTR 3276 exhibited greater susceptibility to culture filtrates SKRU-01 compared to strain PSRDC-4. Furthermore, the culture filtrates SKRU-01 exhibited remarkable thermal stability at 121 °C, degrading AFB1 to 63.91 %. Application of heat-stable culture filtrates SKRU-01 in peanuts demonstrated that the reduction in fungal population and AFB1 production of both pathogenic strains depended significantly (p < 0.05) on the level of heat treatment. The non-treated and 40 °C treated culture filtrates SKRU-01 could reduce AFB1 production to lower than the Standard Aflatoxin Limitation (<20 μg/kg), ensuring food safety and mitigating the health risks associated with aflatoxin exposure.

Improvement of aflatoxin B1 degradation ability by Bacillus licheniformis CotA-laccase Q441A mutant

Aflatoxin B1 (AFB1) contamination seriously threatens nutritional safety and common health. Bacterial CotA-laccases have great potential to degrade AFB1 without redox mediators. However, CotA-laccases are limited because of the low catalytic activity as the spore-bound nature. The AFB1 degradation ability of CotA-laccase from Bacillus licheniformis ANSB821 has been reported by a previous study in our laboratory. In this study, a Q441A mutant was constructed to enhance the activity of CotA-laccase to degrade AFB1. After the site-directed mutation, the mutant Q441A showed a 1.73-fold higher catalytic efficiency (kcat/Km) towards AFB1 than the wild-type CotA-laccase did. The degradation rate of AFB1 by Q441A mutant was higher than that by wild-type CotA-laccase in the pH range from 5.0 to 9.0. In addition, the thermostability was improved after mutation. Based on the structure analysis of CotA-laccase, the higher catalytic efficiency of Q441A for AFB1 may be due to the smaller steric hindrance of Ala441 than Gln441. This is the first research to enhance the degradation efficiency of AFB1 by CotA-laccase with site-directed mutagenesis. In summary, the mutant Q441A will be a suitable candidate for highly effective detoxification of AFB1 in the future.

Biodegradation of aflatoxin B1 by Aspergillus terreus HNGD-TM15 and its degradation mechanism

Abstract The Aspergillus terreus strain HNGD-TM15 that was isolated from soil grown with rosemary was shown to efficiently degrade aflatoxin B1 (AFB1). Specifically, AFB1 was degraded by the strain’s fermentation broth at a maximum degradation rate of 98.3%. HPLC and LC-MS analyses detected a degradation product with an m/z ion value of 312.0636. Its molecular formula was C18H39NO3, which was tentatively identified as 2-amino-1, 3,4-octadecatriol. Based on LC-MS results and further analysis, it was revealed that a series of reactions, such as decomposition, reduction (lactone ring hydrogenation) and substitution (hydrolysis), occurred during the degradation of AFB1. Therefore, A. terreus HNGD-TM15 has a great potential for application in the detoxification of AFB1 contaminating food and feed products.

Novel ZIF-8/ZnS hollow polyhedral heterostructures derived from ZIF-8 with enhanced photocatalytic activity for degradation of aflatoxin B1

Mycotoxins, formed in lots of foods during the production, processing and transportation processes, are the secondary metabolites of Fusarium genus, Penicillium and Aspergillus. Aflatoxin B1 (AFB1) is considered as one of the most dangerous mycotoxins for human and animals due to its strong carcinogenicity and hepatotoxic effects. Hence, degradation of AFB1 completely or reduction of AFB1 to a low degree content has caused much attentions. In this paper, novel ZIF-8/ZnS hollow polyhedral heterostructures were successfully synthesized by sulphurizing ZIF-8 via solvothermal method. The synthesized samples were used as catalysts for AFB1 degradation under UV light irradiation. All as-synthesized samples were analyzed by XRD, SEM, BET, UV–vis, ESR and so on. The results show that parts of ZIF-8 can be sulphurized by TAA to synthesize ZnS and the ZIF-8 particles were etched to form the hollow structure. The sulphurizing degree of ZIF-8 has great influence on the photocatalytic activity of ZIF-8/ZnS. Compared with ZIF-8/ZnS (50) and ZIF-8/ZnS (150), ZIF-8/ZnS (100) exhibited the best photocatalytic activity and could degrade 98.6 ​% of AFB1. Except that, the reactive species of ZIF-8/ZnS (100) were confirmed to be ·OH and ·O2−. This strategy of coupling MOFs with semiconductor provides a facile method to synthesize for catalyst with enhanced photocatalytic activity for degradation of food pollutants produced by aflatoxins. KeyworksAflatoxin B1, ZIF-8/ZnS, photocatalytic activity, reactive species.

Optimizing the degradation of aflatoxin B1 in corn by Trametes versicolor and improving the nutritional composition of corn.

Corn, being an important grain, is prone to contamination by aflatoxin B1 (AFB1 ), and AFB1 -contaminated corn severely endangers the health of humans and livestock. Trametes versicolor, a fungus that can grow in corn, possesses the ability to directly degrade AFB1 through its laccase. This study aimed to optimize the fermentation conditions for T. versicolor to degrade AFB1 in corn and investigate the effect of T. versicolor fermentation on the nutritional composition of corn. AFB1 -contaminated corn was used as the culture substrate for T. versicolor. A combination of single-factor experiments and response surface methodology was employed to identify the optimal conditions of AFB1 degradation. The optimal conditions of AFB1 degradation were as follows: 9 days of fermentation, a fermentation temperature of 26.7 °C, a moisture content of 70.5% and an inoculation amount of 4.9 mL (containing 51.99 mg of T. versicolor mycelia). With the optimal conditions, the degradation rate of AFB1 in corn could reach 93.01%, and the dry basis content of protein and dietary fiber in the fermented corn was significantly increased. More importantly, the lysine content in the fermented corn was also significantly increased. This is the first report that direct fermentation of AFB1 -contaminated corn by T. versicolor not only efficiently degrades AFB1 but also improves the nutritional composition of corn. These findings suggest that the fermentation of corn by T. versicolor is a promising, environmentally friendly and efficient approach to degrade AFB1 and improve the nutritional value of corn. © 2023 Society of Chemical Industry.

Non-thermal plasma inhibited the growth and aflatoxins production of Aspergillus flavus, degraded aflatoxin B1 and its potential mechanisms

Aspergillus flavus (A. flavus) and its toxic metabolites, aflatoxins, are widely distributed in agricultural products and foods, posing a grave threat to food safety and public health. This study investigated the decontamination effects and mechanism of non-thermal plasma (NTP) on A. flavus and aflatoxins. NTP primarily generated reactive oxygen and nitrogen species (RONS), effectively inactivating A. flavus spores via the Weibull + Tail model. The results of ultrastructural observation and membrane integrity revealed that fungal inactivation by NTP was mainly attributed to intracellular destruction rather than the direct damage of cell membranes. Furthermore, NTP induced the accumulation of intracellular ROS and NO, disrupting anti-oxidative and energy metabolic systems, thereby inducing fungal apoptosis via a caspase-dependent mitochondrial pathway, which is responsible for A. flavus spore inactivation. In addition, the results showed that NTP reduced aflatoxin B1 (AFB1) production by regulating the expression of key genes in the aflatoxin biosynthetic pathway. Moreover, NTP effectively degraded AFB1 by disintegrating the furan ring, modifying the methoxy group, and forming the double bond in cyclopentanone, thereby decreasing the toxicity of AFB1. Hence, NTP exhibits exceptional decontamination effects against A. flavus and AFB1 via fungal inactivation, aflatoxin biosynthesis suppression, and AFB1 degradation, making it a viable alternative strategy for agricultural disinfection.