Patulin Degradation Research Articles

The transcriptomic and proteomic changes of Hannaella sinensis in response to patulin stress

ABSTRACT Patulin (PAT) is a significant mycotoxin contaminant of fruit and fruit products. Hannaella sinensis is an antagonistic yeast that controls postharvest diseases and mycotoxins in crops and is potentially valuable for ensuring food safety. Our previous studies have shown that H. sinensis can effectively degrade PAT. The possible physiological mechanisms of yeast have been previously reported, however, the molecular mechanism of PAT degradation by the yeast needs to be explored in depth. Therefore, our current work aimed to use transcriptomic and proteomic techniques to unravel the molecular mechanisms by which PAT degradation occurs in H. sinensis. The results showed that transcriptomic studies of H. sinensis in PAT culture identified 951 differentially expressed genes, of which 194 were up-regulated, and 757 were down-regulated. These genes could be used to induce the ability of H. sinensis to resist stress, promote the synthesis of drug-resistant substances and maintain the integrity of cell membranes. Besides, proteomic analyses revealed that PAT treatment enhances multiple aspects of H. sinensis, including protein processing, cell growth, reproduction, catalase, cell wall synthesis, and more. Our study helps to understand the possible mechanisms of PAT degradation and PAT stress response at the transcription and protein expression levels in H. sinensis.

Effect of patulin on the growth and nutrient composition of black soldier fly larvae

Abstract Black solider fly (BSF, Hermetia illucens) is a common insect in most part of North America, Europe, and Asia, and it is one of the most promising alternative protein sources for animal consumption. BSF can be fed on food waste and agricultural byproducts, which are usually contaminated with mycotoxins. Therefore, it is important to study the fate of mycotoxins in BSF and how mycotoxins will affect the growth and nutrient composition of BSF. Patulin is one of the major mycotoxins found in rotten fruits and consumption of patulin contaminated food can cause both long term and short-term health problems in both human and livestock. In this study clean and patulin spiked apple puree were provided to 50 three days old black soldier fly larvae (BSFL) for 3, 6, 9 and 17 days. Patulin bioaccumulation and degradation by BSFL, survival rate, weight gain and nutrient composition of BSFL were evaluated. During the treatment period, no patulin bioaccumulation was found in BSFL. BSFL was able to degrade patulin and convert patulin into (E)-ascladiol. By day 17, BSFL degraded 89.7% of patulin, 5.7% of patulin was converted into (E)-ascladiol, and 10.3% of patulin remained in its original form. While patulin treatment led to lower biomass and growth rate in BSFL, greater protein content and lower lipid content was observed in patulin treated BSFL. Thus, we conclude that BSFL do not accumulate patulin, and it is able to degrade patulin and converts patulin into its derivatives. The applied patulin negatively affects the development of BSLF resulting in lower biomass, growth rate and lipid content, but it induces higher protein content in BSFL.

Current insights into yeast application for reduction of patulin contamination in foods: A comprehensive review.

Patulin, a fungal secondary metabolite with multiple toxicities, is widely existed in a variety of fruits and their products. This not only causes significant economic losses to the agricultural and food industries but also poses a serious threat to human health. Conventional techniques mainly involved physical and chemical methods present several challenges include incomplete patulin degradation, high technical cost, and fruit quality decline. In comparison, removal of mycotoxin through biodegradation is regarded as a greener and safer strategy which has become popular research. Among them, yeast has a unique advantage in detoxification effect and application, which has attracted our attention. Therefore, this review provides a comprehensive account of the yeast species that can degrade patulin, degradation mechanism, current application status, and future challenges. Yeasts can efficiently convert patulin into nontoxic or low-toxic substances through biodegradation. Alternatively, it can use physical adsorption, which has the advantages of safety, high efficiency, and environmental friendliness. Nevertheless, due to the inherent complexity of the production environment, the sole utilization of yeast as a control agent remains inherently unstable and challenging to implement on a large scale in a practical manner. Integration control, enhancement of yeast resilience, improvement of yeast cell wall adsorption capacity, and research on additional patulin-degrading enzymes will facilitate the practical application of this approach. Furthermore, we analyzed the feasibility of the yeast commercial application in patulin reduction and provided suggestions on how to enhance its commercial value, which is of great significance for the control of mycotoxins in food products.

Identification and application of a novel patulin degrading enzyme from Cyberlindnera fabianii

Patulin (PAT) is a mycotoxin commonly found in fruits and vegetables, prompting the need for effective removal and detoxification methods, which have garnered significant research attention in recent years. Among these methods, the utilization of microbial-derived enzymes stands out due to their mild operating conditions, specificity in targeted functional groups, and the production of non-toxic by-products, making it a preferred degradation approach. In this study, a novel PAT-degrading enzyme derived from Cyberlindnera fabianii (Cyfa-SDR) was identified, demonstrating its highest catalytic activity at pH 7.0 and 80 °C against PAT. This temperature tolerance level represents the highest reported for PAT-degrading enzymes to date. The enzyme was further characterized as a short-chain dehydrogenase through analysis of its amino acid composition, conserved GXXXGXG motif, and dependency on NADPH. Moreover, the study evaluated the efficiency of PAT degradation by Cyfa-SDR at varying substrate and enzyme concentrations, surpassing the performance of other PAT-degrading enzymes, thus highlighting its substantial potential for the biological control of PAT. In conclusion, the enzymatic treatment using the PAT-degrading enzyme Cyfa-SDR presents a viable and promising solution for enhancing the quality and safety of fruit juice.

Patulin Biodegradation Mechanism Study in Pichia guilliermondii S15-8 Based on PgSDR-A5D9S1.

Patulin contamination has become a bottleneck problem in the safe production of fruit products, although biodegradation technology shows potential application value in patulin control. In the present study, the patulin biodegradation mechanism in a probiotic yeast, Pichia guilliermondii S15-8, was investigated. Firstly, the short-chain dehydrogenase PgSDR encoded by gene A5D9S1 was identified as a patulin degradation enzyme, through RNA sequencing and verification by qRT-PCR. Subsequently, the exogenous expression system of the degradation protein PgSDR-A5D9S1 in E. coli was successfully constructed and demonstrated a more significant patulin tolerance and degradation ability. Furthermore, the structure of PgSDR-A5D9S1 and its active binding sites with patulin were predicted via molecular docking analysis. In addition, the heat-excited protein HSF1 was predicted as the transcription factor regulating the patulin degradation protein PgSDR-A5D9S1, which may provide clues for the further analysis of the molecular regulation mechanism of patulin degradation. This study provides a theoretical basis and technical support for the industrial application of biodegradable functional strains.

Study on the mechanism of photocatalytic degradation of patulin in simulated apple juice.

Patulin poses a potential risk to human health, and current methods for removing it have certain limits. Thus, the effective and secure technique for degrading patulin in juice is critical. In this study, a nitrogen-doped chitosan-TiO2 nanocomposite (N-TiO2 Nps) as a photocatalyst was employed to decompose patulin. Under the action of the photocatalyst, 500μg/L patulin was completely degraded within 1h in simulated juice. Quenching experiments identified superoxide and hydroxyl radicals as the dominant species responsible for patulin degradation. On the bases of liquid chromatography-mass spectrometry (LC-MS) and density functional theory (DFT) calculation, the reaction sites in patulin were predicted and a possible photodegradation pathway was proposed. The findings not only elucidated a new method for removing patulin but also provided a theoretical basis for scrutinizing the degradation mechanism of mycotoxins.

New insights into searching patulin degrading enzymes in Saccharomyces cerevisiae through proteomic and molecular docking analysis

Global warming has increased the contamination of mycotoxins. Patulin (PAT) is a harmful contaminant that poses a serious threat to food safety and human health. Saccharomyces cerevisiae biodegrades PAT by its enzymes during fermentation, which is a safe and efficient method of detoxification. However, the key degradation enzymes remain unclear. In this study, the proteomic differences of Saccharomyces cerevisiae under PAT stress were investigated. The results showed that the proteins involved in redox reactions and defense mechanisms were significantly up-regulated to resist PAT stress. Subsequently, molecular docking was used to virtual screen for degrading enzymes. Among 18 proteins, YKL069W showed the highest binding affinity to PAT and was then expressed in Escherichia coli, where the purified YKL069W completely degraded 10 μg/mL PAT at 48 h. YKL069W was demonstrated to be able to degrade PAT into E-ascladiol. Molecular dynamics simulations confirmed that YKL069W was stable in catalyzing PAT degradation with a binding free energy of − 7.5 kcal/mol. Furthermore, it was hypothesized that CYS125 and CYS101 were the key amino acid residues for degradation. This study offers new insights for the rapid screening and development of PAT degrading enzymes and provides a theoretical basis for the detoxification of mycotoxins.

Overexpression of the SDR gene improves the ability of Meyerozyma guilliermondii to degrade patulin in pears and juices.

The intracellular enzymes of antagonistic yeast are effective in controlling patulin (PAT) contamination. However, countless enzymes that have been revealed remain functionally uncharacterized. The study built on previous transcriptomic data obtained by our research group to amplify and express a gene encoding a short-chain dehydrogenase/reductase (SDR) in Meyerozyma guilliermondii. Overexpression of SDR increased the tolerance of M. guilliermondii to PAT and the ability to degrade PAT of the intracellular enzymes. Furthermore, MgSDR-overexpressed M. guilliermondii showed higher PAT degradation in juices (apple and peach) and controlled the blue mold of pears at 20°C and 4°C while significantly reduced the content of PAT and the biomass of Penicillium expansum in decayed tissues than wild-type M. guilliermondii. This study provides theoretical references for the subsequent heterologous expression, formulation, and application of the SDR protein from M. guilliermondii and contributes to elucidating the PAT degradation mechanism of antagonistic yeasts.

MOF-derived La–ZnFe2O4@Fe3O4@carbon magnetic hybrid composite as a highly efficient and recyclable photocatalyst for mycotoxins degradation

During the planting, storage, or feeding stages, foodstuffs can be easily polluted by mycotoxins such as aflatoxin B1, patulin and zearalenone, which are produced by pathogenic microorganisms. Therefore, the removal of these mycotoxins constitutes an active area of research. Herein, we used an Fe/Zn metal–organic framework (MOF) as a precursor to fabricate a novel La–ZnFe2O4@Fe3O4@carbon magnetic hybrid composite via a simple absorption–pyrolysis method and explored its photocatalytic performance in the degradation of aflatoxin B1, patulin, and zearalenone. La–ZnFe2O4@Fe3O4@carbon can effectively degrade 98.37% of aflatoxin B1, outperforming Fe3O4@carbon (69.57%) and ZnFe2O4@Fe3O4@carbon (91.39%). Moreover, La–ZnFe2O4@Fe3O4@carbon exhibits superior stability and excellent photocatalytic activity for the degradation of patulin (97.35%) and zearalenone (98.52%). Transient photocurrent response measurements reveal that the La doping and the combination with ZnFe2O4 highly promote the separation of photoinduced electron–hole pairs on Fe3O4@carbon. An electron spin resonance analysis suggests that ·OH and ·O2− are the main active radicals during the photodegradation process. Thus, a plausible photodegradation pathway for the degradation of mycotoxins produced by pathogenic microorganisms is proposed.

Molecular Response of Meyerozyma guilliermondii to Patulin: Transcriptomic-Based Analysis

Patulin (PAT), mainly produced by Penicillium expansum, is a potential threat to health. In recent years, PAT removal using antagonistic yeasts has become a hot research topic. Meyerozyma guilliermondii, isolated by our group, produced antagonistic effects against the postharvest diseases of pears and could degrade PAT in vivo or in vitro. However, the molecular responses of M. guilliermondii over PAT exposure and its detoxification enzymes are not apparent. In this study, transcriptomics is used to unveil the molecular responses of M. guilliermondii on PAT exposure and the enzymes involved in PAT degradation. The functional enrichment of differentially expressed genes indicated that the molecular response mainly includes the up-regulated expression of genes related to resistance and drug-resistance, intracellular transport, growth and reproduction, transcription, DNA damage repair, antioxidant stress to avoid cell damage, and PAT detoxification genes such as short-chain dehydrogenase/reductases. This study elucidates the possible molecular responses and PAT detoxification mechanism of M. guilliermondii, which could be helpful to further accelerate the commercial application of antagonistic yeast toward mycotoxin decontamination.

Detoxification of patulin in apple juice by enzymes and evaluation of its degradation products

The contamination of patulin is widely present in apple, hawthorn, pear, and their products. Many patulin detoxification methods have already been reported, among which enzymes are considered as one of the most promising degradation methods. However, in the reports available so far, patulin-degrading enzymes have rarely been separated and produced, making their application limited. In this study, four commercial enzymes including esterase, peroxidase, lipase A, and lipase M with the potential ability were used for patulin degradation. The results showed that four enzymes can degrade patulin with a rate of 97.8%, 53.2%, 76.3%, and 68.2%, respectively. The soluble solids content, pH value, browning degree, transmittance, organic acids and soluble sugar content, and volatile components of apple juice did not vary much or were within the acceptable range. The main degradation products of patulin were desoxypatulinic acid and hydroascladiol, the toxicity of which have been reported to be lower than that of patulin. Our discovery may provide a more convenient option for enzymes in juice detoxification. • The removal effects of four commercial enzymes on patulin were evaluated. • Esterase with a high degradation rate of 97.8% for Patulin was screened. • The effects of commercial enzymes on the quality of apple juice were evaluated. • The degradation products DPA and Hydroascladiol with lower toxicity were obtained.

Role of thiols and ascladiol production in patulin degradation by lactobacilli

Patulin is a mycotoxin contaminant in various foods with apple products being its major dietary source. Yeast can reduce patulin levels during fermentation via biotransformation and thiol-adduct formation, with the ability of patulin to react with thiols being well known. Conversion of patulin to ascladiol by lactobacilli has been sparsely reported, while the contribution of thiols in reduction of patulin levels by lactobacilli remains undocumented. In this study, 11 strains of lactobacilli were screened for ascladiol formation in apple juice fermentation. Highest bioconversion was obtained for Lactiplantibacillus plantarum strains followed by Levilactobacillus brevis TMW1.465. Ascladiol production was also detected in several other lactobacilli species albeit in trace amounts. Reduction in patulin levels by Fructilactobacillus sanfranciscensis DMS 20451 and its glutathione reductase (ΔgshR) negative mutant was also assayed to determine the contribution of thiols. The hydrocinnamic acid reductase of Furfurilactobacillus milii did not contribute to reduction of patulin levels. In conclusion, this study demonstrated the potential of various lactobacilli in reduction of patulin levels via biotransformation of patulin to ascladiol, while also providing evidence for the role of thiol formation by lactobacilli and its presence in reducing patulin levels during fermentation.

Efficient Biodegradation of Patulin by Aspergillus niger FS10 and Metabolic Response of Degrading Strain.

Patulin, a mycotoxin commonly found in fruits and derived products, causes serious health problems for humans and animals worldwide. Several microbial strains have been observed to possess the ability to effectively remove patulin. However, these methods are presently associated with disadvantages such as low degradation efficiency and an unclear biodegradation mechanism. In the current study, the characteristics of patulin degradation via Aspergillus niger FS10 were evaluated, and the mechanisms involved were analyzed using metabolomics technologies. The results showed that the suspension of A. niger FS10 could degrade 94.72% of patulin within 36 h. The moment concentration pf patulin was 0.116 μg/mL, and the detection limit value was 0.01 μg/mL. In addition, the patulin content was reduced to levels below the detection limit within 48 h. A. niger FS10 mainly degrades patulin by producing intracellular enzymes, which can convert patulin into ascladiol. This degradation method can effectively reduce the damage caused by patulin to HepG2 cells. In addition, the patulin treatment significantly affects the pentose phosphate pathway and the glutathione pathway. These two metabolic pathways are speculated to be closely related to patulin degradation via A. niger FS10. The incubation of A. niger FS10 with patulin-contaminated apple pomace can not only eliminate patulin but also increase the utilization of apple pomace. Therefore, our research results provide a new method for addressing patulin contamination in the food and feed industries.

Degradation of Patulin in Apple Juice by Pulsed Light and its Effect on the Quality

Degradation of Patulin in Apple Juice by Pulsed Light and its Effect on the Quality

Ultraviolet Applications to Control Patulin Produced by Penicillium expansum CMP-1 in Apple Products and Study of Further Patulin Degradation Products Formation and Toxicity

Patulin is a mycotoxin whose presence in apple-derived products and fruit juices is legally regulated, being its maximum limits established in the legislation of multiple countries. However, the management of contaminated batches is still an issue for producers. This investigation aims to evaluate ultraviolet light (254 nm, UV-C254nm) irradiation to find solutions that can be applied at different stages of the apple juice production chain. In this regard, 8.8 (UV-1) and 35.1 (UV-2) kJ m−2 treatments inactivated spores of Penicillium expansum CMP-1 on the surface of apples. Although the same treatments applied to wounded apples (either before the infection or after the infection, immediately or when the lesion had appeared) did not show any effect on the growth rate of P. expansum during storage (up to 14 days, at 4 or 25 °C), they reduced patulin content per lesion size in apples treated after the infection had occurred (patulin decreased from 2.24 (control) to 0.65 µg kg−1 cm−2 (UV-2 treated apples)). Additionally, the treatment of juice with patulin with ultraviolet light up to 450.6 kJ m−2 resulted in more than 98 % reduction of patulin. Degradation products of patulin after UV-C254nm treatments were tentatively identified by HPLC–MS, and toxicity and biological activities were assessed in silico, and results indicated that such products did not pose an increased risk when compared to patulin.

Effect of the antagonistic yeast Hannaella sinensis on the degradation of Patulin

Patulin (PAT) is a toxic fungal metabolite that is widely distributed in food and poses a serious health risk to humans and animals. Hannaella sinensis has been found to be effective in controlling post-harvest diseases of apples. However, it is not known whether H. sinensis can degrade PAT. The aim of this study was to investigate the degradation efficacy of PAT by H. sinensis and the possible mechanisms involved. The experimental results showed that H. sinensis degraded 100 % of PAT in NYDB medium within 30 h. The living cells of H. sinensis did not absorb PAT, and the cell wall and extracellular metabolites of H. sinensis were not involved in the degradation of PAT. The addition of cycloheximide to the culture medium inhibited the degradation of PAT by H. sinensis. Our results indicated that H. sinensis could degrade PAT and that the intracellular enzymes of H. sinensis played a crucial role in the degradation process. This result provides a new solution for the degradation of PAT.

Alterations in the proteome as a regulating mechanism for patulin stress by the antagonistic yeast Meyerozyma guilliermondii

Meyerozyma guilliermondii is an antagonistic yeast; our research team has recently reported its biocontrol activities against blue mold decay of pears and strong patulin (PAT) reduction ability in vitro. PAT is a harmful mycotoxin produced by some species of Penicillium, Aspergillus, and Byssochlamys. The PAT degradation process and molecular mechanism(s) of M. guilliermondii remain unclear. The data-independent acquisition (DIA) strategies promise a comprehensive, reproducible, digital, qualitative and quantitative record of the proteins present in a sample. Differentially expressed proteins accompanying PAT degradation in M. guilliermondii were observed to investigate PAT reduction mechanisms. The comparative proteomic analysis of M. guilliermondii showed that 136 proteins were up-regulated and 122 were down-regulated upon PAT degradation. The M. guilliermondii's regulation of PAT response mainly includes the up-regulation of (1) growth and reproduction-related protein expression, (2) transcription and translation-related protein expression, (3) resistance-related protein expression, (4) multidrug transporters, (5) anti-oxidative stress-related proteins, (6) the cell wall and cell membrane production-related proteins, and (7) PAT degradation-related protein expression. This paper analyzed M. guilliermondii’s molecular response and regulation upon PAT reduction at the gene and protein expression levels. Our results provide a solid foundation for PAT control in fruits and their products by antagonistic yeasts.

Degradation of Patulin in Pear Juice and Apple Juice by Ascorbic Acid and the Combination of Ascorbic Acid and Ferrous Iron.

Patulin (PAT) is a toxic secondary metabolite produced by certain species of Penicillium sp. and Aspergillus sp. on apples and pears. In this study, we investigated the effects of ascorbic acid and the combination of ascorbic acid and ferrous iron on degradation of PAT in 100% pure pear juice and apple juice using high-performance liquid chromatography UV detector (HPLC-UVD). The addition of 2 different levels of ascorbic acid (143 or 286 μg/mL) into pear juice or apple juice containing 0.08 or 0.4 μg/mL of PAT showed 87.7-100% and 67.3-68.7% of PAT degradation rates, respectively, after 24 h incubation at 25 °C. Moreover, the addition of both ascorbic acid (143 or 286 μg/mL) and ferrous iron (0.033 or 0.11 μmol/mL) into pear juice or apple juice containing the same level of PAT exhibited higher PAT degradation rates (100 and 75-94%, respectively) than the addition of only ascorbic acid after 24 h incubation at 25 °C. Our data demonstrated that ascorbic acid plus ferrous iron as well as ascorbic acid were highly effective on degradation of PAT in pear juice and apple juice and that addition of both ascorbic acid and ferrous iron produced higher PAT degradation rates than addition of only ascorbic acid.

Possible Reaction Mechanisms Involved in Degradation of Patulin by Heat-Assisted Cysteine under Highly Acidic Conditions

Patulin (PAT) is one of mycotoxins that usually contaminates apple juice, and it is not easily detoxified by cysteine (CYS) at room temperature due to the highly acidic conditions based on the Michael addition reaction. However, it could be effectively degraded by a heating treatment at 120 °C for 30 min in the presence of cysteine. In our study, a total of eight degradation products (DP A–H) were characterized and identified via liquid chromatography quadrupole time-of-flight mass spectrometry (LC-Q-TOF-MS) in a negative ion mode, and their structures and formulas were proposed based on their accurate mass data. The fragmentation patterns of PAT and its degradation products were obtained from the MS/MS analysis. Meanwhile, the possible reaction mechanisms involved in the degradation of PAT were established and explained for the first time. According to the relation between the structure and toxicity of PAT, it could be deduced that the toxic effects of PAT degradation products were potentially much less than those of PAT-self.

Insights into the Metabolic Response of Lactiplantibacillus plantarum CCFM1287 upon Patulin Exposure.

Patulin (PAT) is a common mycotoxin in the food industry, and is found in apple products in particular. Consumption of food or feed contaminated with PAT can cause acute or chronic toxicity in humans and animals. Lactiplantibacillus plantarum CCFM1287 is a probiotic strain that effectively degrades PAT in PBS and food systems. In this study, it was found that the concentration of PAT (50 mg/L) in MRS medium decreased by 85.09% during the first stages of CCFM1287 growth, and this change was consistent with the first-order degradation kinetic model. Meanwhile, the regulation of oxidative stress by L. plantarum CCFM1287 in response to PAT exposure and metabolic changes that occur during PAT degradation were investigated. The degree of intracellular damage was attenuated after 16 h of exposure compared to 8 h. Meanwhile, metabolomic data showed that 30 and 29 significantly different metabolites were screened intracellularly in the strain after 8 h and 16 h of PAT stress at 50 mg/L, respectively. The results of pathway enrichment analysis suggested that the purine metabolic pathway was significantly enriched at both 8 h and 16 h. However, as is consistent with the performance of the antioxidant system, the changes in Lactiplantibacillus diminished with increasing time of PAT exposure. Therefore, this study helps to further explain the mechanism of PAT degradation by L. plantarum CCFM1287.