Patulin is a toxic chemical contaminant produced by several species of mold, especially within Aspergillus, Penicillium and Byssochlamys. It is the most common mycotoxin found in apples and ...apple-derived products such as juice, cider, compotes and other food intended for young children. Exposure to this mycotoxin is associated with immunological, neurological and gastrointestinal outcomes. Assessment of the health risks due to patulin consumption by humans has led many countries to regulate the quantity in food. A full understanding of the molecular genetics of patulin biosynthesis is incomplete, unlike other regulated mycotoxins (aflatoxins, trichothecenes and fumonisins), although the chemical structures of patulin precursors are now known. The biosynthetic pathway consists of approximately 10 steps, as suggested by biochemical studies. Recently, a cluster of 15 genes involved in patulin biosynthesis was reported, containing characterized enzymes, a regulation factor and transporter genes. This review includes information on the current understanding of the mechanisms of patulin toxinogenesis and summarizes its toxicological effects.
•Degradation of patulin in apple juices was achieved by pulsed light processing.•Supplementation with glutathione and ferrous ions enhanced patulin degradation.•Glutathione-patulin adducts were ...identified as patulin degradation products.
Patulin is a toxic mycotoxin usually associated with apple products. Due to its unhealthy effects for humans, its content is regulated by the food safety authorities. The removal or degradation of this mycotoxin in contaminated apple juices has been studied with different approaches with uneven effectiveness. However, a strategy based on the chemical reaction between patulin and glutathione (GSH), in order to generate the conjugates that are formed during cell detoxification process, is an innovative approach yet to be evaluated. In this work, the formation of patulin-GSH conjugates activated by the application of pulsed light treatments and catalyzed by Fe2+ ions was evaluated. The study of patulin degradation and effect of the GSH/Fe2+ molar ratio showed that a molar ratio of 5 allows an adequate catalytic effect of the metal ions. In addition, mono-substituted patulin-glutathione adducts were identified as the main type of generated conjugates.
•A novel UiO-66(NH2)@Au-Cys adsorbent is applied to patulin removal for the first time.•UiO-66(NH2)@Au-Cys exhibits high patulin adsorption capacity.•UiO-66(NH2)@Au-Cys has potential applications in ...the apple juice industry for patulin removal.
Patulin (PAT) is one of the most common toxic contaminants of apple juice, which causes severe food safety issues throughout the apple industry. In order to remove PAT efficiently, a metal-organic framework-based adsorbent (UiO-66(NH2)@Au-Cys) was successfully synthesized and used for PAT removal from juice-pH simulation solution and real apple juice. Batch adsorption experiments were systematically performed to study the adsorption behavior for PAT. The results showed that adsorption process could be well described by the Pseudo-second order model and Freundlich isotherm model. The maximum adsorption capacity (4.38 µg/mg) was 10 times higher than the microbe-based biosorbents. Thermodynamic investigation demonstrated that adsorption process was spontaneous and endothermic. Furthermore, no marked cytotoxicity on NIH 3T3 cell lines was observed when the concentration of the adsorbent was lower than 10 μg/mL. Therefore, UiO-66(NH2)@Au-Cys is a potential adsorbent for PAT removal from apple juice with little quality changes.
Summary
The patulin biosynthesis is one of model pathways in an understanding of secondary metabolite biology and network novelties in fungi. However, molecular regulation mechanism of patulin ...biosynthesis and contribution of each gene related to the different catalytic enzymes in the biochemical steps of the pathway remain largely unknown in fungi. In this study, the genetic components of patulin biosynthetic pathway were systematically dissected in Penicillium expansum, which is an important fungal pathogen and patulin producer in harvested fruits and vegetables. Our results revealed that all the 15 genes in the cluster are involved in patulin biosynthesis. Proteins encoded by those genes are compartmentalized in various subcellular locations, including cytosol, nucleus, vacuole, endoplasmic reticulum, plasma membrane and cell wall. The subcellular localizations of some proteins, such as PatE and PatH, are required for the patulin production. Further, the functions of eight enzymes in the 10‐step patulin biosynthetic pathway were verified in P. expansum. Moreover, velvet family proteins, VeA, VelB and VelC, were proved to be involved in the regulation of patulin biosynthesis, but not VosA. These findings provide a thorough understanding of the biosynthesis pathway, spatial control and regulation mechanism of patulin in fungi.
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•A short-chain dehydrogenase/reductase CgSDR has been expressed and purified.•CgSDR is a new patulin degradation enzyme and transformed patulin into E-ascladiol.•VAL188 of CgSDR may ...play a crucial role during the degrading reaction.•CgSDR has potential application for removal of patulin in the apple juice industry.
Biodegradation based on microbial enzymes is considered to be one of the promising ways for controlling patulin contamination. However, few patulin degrading enzymes have been isolated and characterized until now. Here, a short-chain dehydrogenase/reductase (SDR) gene, CgSDR, was cloned from a yeast strain Candida guilliermondii, and expressed in Escherichia coli. The expression of CgSDR conferred a strong patulin tolerance and degradation ability to E. coli, and purified CgSDR could transform patulin into E-ascladiol in vitro with NADPH as a coenzyme. Moreover, addition of CgSDR at 150 μg/mL could reduce 80% of patulin in apple juice and the biodegradation process did not affect the quality of the apple juice. A molecular docking analysis and site-directed mutagenesis indicated that CgSDR might interact with patulin via VAL188 as an active binding sites. The findings provide new insights for developing enzymic formulations for mycotoxin detoxification in fruit derived products.
Patulin (PAT) is the most common mycotoxin found in moldy fruits and their derived products, and is reported to cause diverse toxic effects, including hepatotoxicity, nephrotoxicity, cardiotoxicity, ...neurotoxicity, immunotoxicity, gastrointestinal toxicity and dermal toxicity. The cell death induction by PAT is suggested to be a key cellular mechanism involved in PAT-induced toxicities. Accumulating evidence indicates that the multiple forms of cell death are induced in response to PAT exposure, including apoptosis, autophagic cell death, pyroptosis and ferroptosis. Mechanistically, the cell death induction by PAT is associated the oxidative stress induction via reducing the antioxidant capacity or inducing pro-oxidant NADPH oxidase, the activation of mitochondrial pathway via regulating BCL-2 family proteins, the disruption of iron metabolism through ferritinophagy-mediated ferritin degradation, and the induction of the NOD-like receptor (NLR) family pyrin domain containing 3 (NLRP3) inflammasome/caspase-1/gasdermin D (GSDMD) pathway. In this review article, we summarize the present understanding of the cell death induction by PAT, discuss the potential signaling pathways underlying PAT-induced cell death, and propose the issues that need to be addressed to promote the development of cell death-based approach to counteract PAT-induced toxicities.
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•Apoptosis, pyroptosis, ferroptosis and autophagic cell death are induced by patulin.•Patulin induces apoptosis via inducting oxidative stress and activating mitochondrial pathway.•Patulin induces pyroptosis via activating NLRP3/caspase-1/GSDMD pathway.•Patulin induces ferroptosis via inactivating SLC7A11 and disrupting iron metabolism.•Autophagy activation by patulin contributes to the induction of apoptosis, pyroptosis and ferroptosis.
•A rabbit anti-patulin-BSA IgG was developed for the detection of fungal toxin patulin.•The rabbit anti-patulin-BSA IgG was applied to a glass carbon electrode sensor confined with graphene ...oxide.•The developed sensor has linearity with concentration of patulin.•The sensor could detect as low as 5 µg/L of patulin in a few minutes.
The presence of fungal-produced patulin in foods poses a high health risk to people because it can cause neurologic and gastrointestinal illnesses. A glass carbon electrode (GCE) sensor was developed for the rapid and sensitive detection of patulin. Anti-patulin-BSA IgG of a rabbit was produced and immobilised on a GCE coated with a graphene oxide/gold nanocomposite. The mycotoxin patulin in the samples could be captured by the anti-patulin-BSA IgG on the surface of the GCE sensor. The spatial hindrance effect of IgG on the GCE sensor was reduced by the reaction between IgG and patulin, resulting in a decrease in the electron transfer resistance. The current changes in the immobilised anti-patulin-BSA IgG GCE sensor exhibited a linear relationship with patulin concentration and facilitated the sensitive detection of patulin. This immuno-electrochemical GCE sensor could rapidly detect patulin in less than 1 min with a detection limit of 5 µg/L.
Patulin is the main mycotoxin contaminating apples. During the brewing of alcoholic beverages, this mycotoxin is degraded to ascladiol, which is also the last precursor of patulin. The present study ...aims (1) to characterize the last step of the patulin biosynthetic pathway and (2) to describe the toxicity of ascladiol. A
patE
deletion mutant was generated in
Penicillium expansum
. In contrast to the wild strain, this mutant does not produce patulin but accumulates high levels of E-ascladiol with few traces of Z-ascladiol. This confirms that
patE
encodes the patulin synthase involved in the conversion of E-ascladiol to patulin. After purification, cytotoxicities of patulin and E- and Z-ascladiol were investigated on human cell lines from liver, kidney, intestine, and immune system. Patulin was cytotoxic for these four cell lines in a dose-dependent manner. By contrast, both E- and Z-ascladiol were devoid of cytotoxicity. Microarray analyses on human intestinal cells treated with patulin and E-ascladiol showed that the latter, unlike patulin, did not alter the whole human transcription. These results demonstrate that E- and Z-ascladiol are not toxic and therefore patulin detoxification strategies leading to the accumulation of ascladiol are good approaches to limit the patulin risk.
•SD-LLLME is applied to extract patulin for the first time.•ID-LC-MS method is developed for the analysis of patulin in apple juice.•Method is validated with fortified samples.
Quick and simple ...analytical methodology has been developed for the measurement of patulin in apple juice by combining single-drop liquid-liquid-liquid microextraction with isotope dilution ultra-high performance liquid chromatography-mass spectrometry. After systematic parameters optimization, the whole sample pretreatment only consisted of single extraction that consumed 1.5 mL ethyl acetate. While, other methods usually needed complicated pretreatment, including extraction, purification, evaporation and redissolution. This sample pretreatment method could greatly lower the interferences from sugar-rich matrix, and the limit of detection was 0.5 μg/L and limit of quantification was 2 μg/L in apple juice. Moreover, linear range covered three orders of magnitude from 2 to 2000 μg/L. The proposed method is promising for patulin extraction and detection in apple juice, which will opens a new perspective in the enrichment of trace contaminations in high sugar complex matrix.
Mycotoxins can be found in food and feed storage as well as in several kinds of foodstuff and are capable of harming mammals and some of them even in small doses. This study investigated on the ...undifferentiated neuronal cell line SH-SY5Y the effects of two mycotoxins: patulin (PAT) and citrinin (CTN), which are predominantly produced by fungi species Penicillium and Aspergillus. Here, the individual and combined cytotoxicity of PAT and CTN was investigated using the cytotoxic assay MTT. Our findings indicate that after 24 h of treatment, the IC50 value for PAT is 2.01 μM, which decreases at 1.5 μM after 48 h. In contrast, CTN did not attain an IC50 value at the tested concentration. Therefore, we found PAT to be the more toxic compared to CTN. However, the combined treatment suggests an additive toxic effect. With 2,7-dichlorodihydrofluorescin diacetate (DCFH-DA) DCFH-DA assay, ROS generation was demonstrated after CTN treatment, but PAT showed only small changes. The mixture presented a very constant behavior over time. Finally, the median-effect/combination index (CI-) isobologram equation demonstrated an additive effect after 24 h, but an antagonistic effect after 48 h for the interaction of the two mycotoxins.
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