The danger of mycotoxin contamination entering the food supply through post-harvest infection is of perennial concern to food safety experts. To explore the distribution of Penicillium expansum and diffusion of its my...The danger of mycotoxin contamination entering the food supply through post-harvest infection is of perennial concern to food safety experts. To explore the distribution of Penicillium expansum and diffusion of its mycotoxin, patulin, in blue mold-damaged pears, Pyrus bretschneideri Rehd. cv. Yali obtained from markets and orchards in China were artificially inoculated with P. expansum and assayed for patulin accumulation and degree of fungal colonization. The inoculated pears were incubated until the lesions were 5, 10, 20, or 30 mm in diameter. We sampled tissue at a range of distances from the lesion, measured the spread of Penicillium by plate colony-counting methods, and used UHPLC-MS/MS to detect and quantify the patulin concentration. More P. expansum colony-forming units were isolated from pears with a higher degree of decay. Farther from the lesion, the fewer P. expansum colonies were observed, and the lower the patulin content detected. We found a significant difference in the patulin content between samples due to lesion size, and also in tissue sampled 10 mm away from the lesion. In consideration of this finding, to ensure food safety, we recommend that when a blue mold rot lesion on pear is 5, 10, or 20 mm in diameter, 20, 30, and 40 mm beyond the lesion should be removed, respectively. If a lesion surpasses 30 mm in diameter, the whole pear should be thrown away.展开更多
The content of patulin in dairy products was detected by rapid ELISA assay. The pretreatment of samples was simple, with wide range of application. The standard product showed good linear relationship with the range o...The content of patulin in dairy products was detected by rapid ELISA assay. The pretreatment of samples was simple, with wide range of application. The standard product showed good linear relationship with the range of 0-5.4 μg/kg, and the linear relationship reached 0.999 3, with high sensitivity. It took shorter time, with a total time-consumption of 1 h. Moreover, it had good repeatability for analytical requirements.展开更多
An analytical method was developed and validated for determination of patulin in apple puree by HPLC. Extraction and clean-up ofpatulin from clear extract are achieved on AFFINIMIP SPEPATULIN cartridges. Patulin is th...An analytical method was developed and validated for determination of patulin in apple puree by HPLC. Extraction and clean-up ofpatulin from clear extract are achieved on AFFINIMIP SPEPATULIN cartridges. Patulin is then separated on a Hypersil GOLD column 150 mm × 4 mm, 5 μtmand detected at 276 nm. The recovery in the range of 5 μg/kg-80 μg/kg was 81.47%. The limit of detection （LOD） was 1.36 μg/kg, and the limit of quantification （LOQ） was 4.55 μg/kg. The patulin content of the commercial samples of apple puree and samples of apple and fruit puree forinfants and young children as well as the samples of apple puree prepared from two apple varieties intended for processing （Jonathan, Florina） and obtained from conventional and uncertified organic cultures has been evaluated in this paper. The 44.83% patulin concentration of the analyzed samples were under the maximum level of the European Commission Regulation （EC） 1881/2006, in 46.55% of the analyzed samples patulin was not detected and in 8.62% of samples patulin concentration was lower than LOQ （European Comission, 2006a） Patulin was not detected in samples of apple puree intended for infants and young children consumption.展开更多
The mycotoxin, patulin （4-hydroxy-4H-furo [3, 2c] pyran-2 [6H]-one）, is produced by a number of fungi common to fruit and vegetable-based products, most notably apples. Patulin contamination within apple products po...The mycotoxin, patulin （4-hydroxy-4H-furo [3, 2c] pyran-2 [6H]-one）, is produced by a number of fungi common to fruit and vegetable-based products, most notably apples. Patulin contamination within apple products poses a serious health risk to consumers. Studies done on laboratory animals have demonstrated that patulin has a broad spectrum of toxicity, including mutagen city and carcinogenicity. The aim of the experiment was studying influence of selectively acting activated carbon powder--Ercarbon SH （Erbsloh, Germany） which is special produced for lowering HMF （hydroxy methyl furfural）, on reduction of patulin content in clear apple juice. Industrial apple row material with some damaged parts was pressed, juice was pasteurized at 95 ℃ during 2 min. After cooling on 55 ℃, enzymatic treated and clarified juice were filtered by 0.45 [am pore sizes membrane filter, Apple clear juice sample was divided for five parts. The samples of apple juice were diluted to 11.5° Brix and contacted with concentrations of 2, 2.5, 3 and 3.5 g/L activated carbon powder for 30 min. After filtration in the experimental samples, putulin was quantitatively determined by HPLC （high performance liquid chromatography with UV） detector at 276 nm. The research revealed that the best results were achieved by treatment with activated carbon in its powder form at concentration of 2.5 g/L with 30 min contact time.展开更多
In this study,the patulin imprinted and the non-imprinted nanoparticles are synthesized by the two-phase mini emulsion polymerization method and characterized by zeta-size analysis,Fourier transform infrared spectrosc...In this study,the patulin imprinted and the non-imprinted nanoparticles are synthesized by the two-phase mini emulsion polymerization method and characterized by zeta-size analysis,Fourier transform infrared spectroscopy,and scanning electron microscopy.Afterwards,the patulin imprinted and the non-imprinted nanoparticles are attached on the surface of surface plasmon resonance(SPR)chips.The patulin imprinted and the non-imprinted SPR nanosensors are characterized by using atomic force microscope,ellipsometer,and contact angle measurements.Kinetic studies for patulin detection are carried out in the concentration range of 0.5nmol-750nmol.The limit of detection and the limit of quantification values are obtained as 0.011 nmol and 0.036 nmol,respectively.In all kinetic analysis,the response time is 13 min for equilibration,adsorption,and desorption cycles.The selectivity studies of the patulin imprinted and the non-imprinted SPR nanosensors are determined in the presence of ochratoxin A and aflatoxin Bl.In order to demonstrate the applicability,validation studies of the patulin imprinted SPR nanosensor are performed by liquid chromatography-tandem mass spectrometry(LC-MS).展开更多
Patulin(PTL,154 Da)is a small molecule toxin produced as a mold metabolite.PTL is unstable in animals and is quickly converted to its metabolites.Thus,few successful antibodies were reported even after extensive effor...Patulin(PTL,154 Da)is a small molecule toxin produced as a mold metabolite.PTL is unstable in animals and is quickly converted to its metabolites.Thus,few successful antibodies were reported even after extensive efforts in hapten design.In this work,a library immobilization strategy was used to obtain DNA aptamers for PTL,where exposure of PTL to live animals or cells is no longer needed.After 20 rounds of selection,the most abundant aptamer named PTL-1 has a dissociation constant(K_(d))of 12.5μM based on isothermal titration calorimetry and 18.4μM based on thioflavin T(ThT)fluorescence spectroscopy.PTL-1 has excellent selectivity against nine other antibiotics.Compared to previously reported aptamers for PTL,PTL-1 is either shorter or has higher affinity.A label-free ThT fluorescence assay based on PTL-1 was developed,which has a detection limit of 0.2μM(detection range 0.2-20μM)for PTL in fruit juices.The PTL-1 aptamer shows resistance to matrix interference and can be applied to the rapid detection of PTL in a blended juice.This study is a nice demonstration of selecting aptamers for a target molecule difficult for the production of antibodies.展开更多
Patulin(PAT)is a toxic fungal metabolite frequently detected in fruits and derived products,posing significant risks to food safety and public health.In this study,we identified and characterized an NADPH-dependent al...Patulin(PAT)is a toxic fungal metabolite frequently detected in fruits and derived products,posing significant risks to food safety and public health.In this study,we identified and characterized an NADPH-dependent aldoketo reductase(LpAKR)from the probiotic strain Lactiplantibacillus plantarum CCFM1287 that efficiently catalyzes the conversion of PAT into the less toxic metabolite E-ascladiol.Functional disruption of the LpAKR gene in L.plantarum significantly reduced PAT-degrading capacity,confirming its essential role in the detoxification process.The recombinant LpAKR protein was expressed and purified,and its enzymatic activity toward PAT was confirmed in vitro.Structural modeling and molecular docking revealed that PAT binds in a hydrophobic pocket formed by residues Trp24,Trp116,Tyr53,Ala52,Leu78,and Met276,stabilized throughπ-πstacking with Trp116 and positioned adjacent to the NADPH nicotinamide ring.His111 and Asp48 were predicted to participate in proton and electron transfer during catalysis,and site-directed mutagenesis(His111D,Trp116A,Met276A)markedly decreased enzymatic activity,validating their catalytic importance.Molecular dynamics simulations further confirmed the role of these residues in substrate binding and structural stability.These findings provide mechanistic insight into PAT degradation by a food-grade aldo-keto reductase and highlight the potential application of L.plantarum CCFM1287 in enzymatic mycotoxin detoxification strategies for the food industry.展开更多
Selenium(Se)is an essential trace element for humans.Inorganic Se is converted into organic Se that is more easily absorbed during the growth of yeast.Patulin(PAT)is a fungal toxin that can cause damage to the intesti...Selenium(Se)is an essential trace element for humans.Inorganic Se is converted into organic Se that is more easily absorbed during the growth of yeast.Patulin(PAT)is a fungal toxin that can cause damage to the intestinal mucosa.Se can alleviate intestinal damage,and this study mainly focuses on the research of Se enrichment in Kluyveromyces marxianus YG-4(SeK)and its repair effect on jejunal injury induced by PAT.In this study,K.marxianus YG-4 could enrich up to 721.05μg/g of total Se content,of which 86.30%was organic Se.In the organic Se of SeK,protein-bound Se,polysaccharide-bound Se,and nucleic acid-bound Se accounted for 83.22%,10.76%,and 2.68%,respectively.Moreover,SeK had an increased amino acid content compared with K.marxianus YG-4.Finally,the morphology and survival ability of K.marxianus YG-4 were not disrupted after Se enrichment,indicating that SeK has the potential to become a probiotic in the food industry.Furthermore,SeK had the ability to alleviate jejunal damage induced by PAT.Specifically,SeK alleviated intestinal tissue lesions and led to an increase in mucin levels.Moreover,SeK promoted the expression of tight junction proteins(claudin-1,occludin,and ZO-1)and inhibited the activation of the NF-κB signaling pathway.Additionally,SeK reduced harmful bacteria including Turicibacter,Anaerotruncus,and Desulfovibrio,and increased beneficial bacteria including Eggerthellaceae,Muribaculaceae,Dubosiella,and short chain fatty acid producing bacteria(Erysipelotrichaceae and Faecalibaculum).In summary,SeK is expected to become a dietary candidate for alleviating the damage caused by mycotoxins.展开更多
Patulin(PAT)is a harmful mycotoxin that poses a serious threat to food safety,highlighting the urgent need for the development of efficient and environmentally friendly detoxification methods.This study identified a n...Patulin(PAT)is a harmful mycotoxin that poses a serious threat to food safety,highlighting the urgent need for the development of efficient and environmentally friendly detoxification methods.This study identified a novel PAT-degrading enzyme,Glutaredoxin GRX3(GRX3)for the first time.Molecular docking results indicated that PAT stably binds to the hydrophobic cavity of GRX3.Key GRX3 residues interact with PAT through hydrogen bonds.Biochemical analyses revealed that GRX3 is highly active at moderate temperatures(28-45℃)and nearneutral pH and functions via glutathione(GSH)-dependent and thiol-mediated mechanisms.LC-MS/MS analysis confirmed that GRX3 catalyzed the PAT ring opening,converting it into E-ascladiol,which exhibited significantly reduced toxicity.In practical applications,GRX3 effectively degrades PAT in both freshly prepared and commercially available fruit juices,achieving nearly 100%degradation within 24 h in fresh juice.Overall,this study reveals a novel GSH-dependent detoxification pathway for PAT and establishes a strong mechanistic and practical application of GRX3 in agricultural products and food safety control.展开更多
基金supported by the Quality and Safety Risk Assessment for Agro-products of China (GJFP 2015014)
文摘The danger of mycotoxin contamination entering the food supply through post-harvest infection is of perennial concern to food safety experts. To explore the distribution of Penicillium expansum and diffusion of its mycotoxin, patulin, in blue mold-damaged pears, Pyrus bretschneideri Rehd. cv. Yali obtained from markets and orchards in China were artificially inoculated with P. expansum and assayed for patulin accumulation and degree of fungal colonization. The inoculated pears were incubated until the lesions were 5, 10, 20, or 30 mm in diameter. We sampled tissue at a range of distances from the lesion, measured the spread of Penicillium by plate colony-counting methods, and used UHPLC-MS/MS to detect and quantify the patulin concentration. More P. expansum colony-forming units were isolated from pears with a higher degree of decay. Farther from the lesion, the fewer P. expansum colonies were observed, and the lower the patulin content detected. We found a significant difference in the patulin content between samples due to lesion size, and also in tissue sampled 10 mm away from the lesion. In consideration of this finding, to ensure food safety, we recommend that when a blue mold rot lesion on pear is 5, 10, or 20 mm in diameter, 20, 30, and 40 mm beyond the lesion should be removed, respectively. If a lesion surpasses 30 mm in diameter, the whole pear should be thrown away.
文摘The content of patulin in dairy products was detected by rapid ELISA assay. The pretreatment of samples was simple, with wide range of application. The standard product showed good linear relationship with the range of 0-5.4 μg/kg, and the linear relationship reached 0.999 3, with high sensitivity. It took shorter time, with a total time-consumption of 1 h. Moreover, it had good repeatability for analytical requirements.
文摘An analytical method was developed and validated for determination of patulin in apple puree by HPLC. Extraction and clean-up ofpatulin from clear extract are achieved on AFFINIMIP SPEPATULIN cartridges. Patulin is then separated on a Hypersil GOLD column 150 mm × 4 mm, 5 μtmand detected at 276 nm. The recovery in the range of 5 μg/kg-80 μg/kg was 81.47%. The limit of detection （LOD） was 1.36 μg/kg, and the limit of quantification （LOQ） was 4.55 μg/kg. The patulin content of the commercial samples of apple puree and samples of apple and fruit puree forinfants and young children as well as the samples of apple puree prepared from two apple varieties intended for processing （Jonathan, Florina） and obtained from conventional and uncertified organic cultures has been evaluated in this paper. The 44.83% patulin concentration of the analyzed samples were under the maximum level of the European Commission Regulation （EC） 1881/2006, in 46.55% of the analyzed samples patulin was not detected and in 8.62% of samples patulin concentration was lower than LOQ （European Comission, 2006a） Patulin was not detected in samples of apple puree intended for infants and young children consumption.
文摘The mycotoxin, patulin （4-hydroxy-4H-furo [3, 2c] pyran-2 [6H]-one）, is produced by a number of fungi common to fruit and vegetable-based products, most notably apples. Patulin contamination within apple products poses a serious health risk to consumers. Studies done on laboratory animals have demonstrated that patulin has a broad spectrum of toxicity, including mutagen city and carcinogenicity. The aim of the experiment was studying influence of selectively acting activated carbon powder--Ercarbon SH （Erbsloh, Germany） which is special produced for lowering HMF （hydroxy methyl furfural）, on reduction of patulin content in clear apple juice. Industrial apple row material with some damaged parts was pressed, juice was pasteurized at 95 ℃ during 2 min. After cooling on 55 ℃, enzymatic treated and clarified juice were filtered by 0.45 [am pore sizes membrane filter, Apple clear juice sample was divided for five parts. The samples of apple juice were diluted to 11.5° Brix and contacted with concentrations of 2, 2.5, 3 and 3.5 g/L activated carbon powder for 30 min. After filtration in the experimental samples, putulin was quantitatively determined by HPLC （high performance liquid chromatography with UV） detector at 276 nm. The research revealed that the best results were achieved by treatment with activated carbon in its powder form at concentration of 2.5 g/L with 30 min contact time.
基金This study was partly supported by Scientific Research Foundation of Hacettepe University Project(Grant No.FHD-2019-18247).
文摘In this study,the patulin imprinted and the non-imprinted nanoparticles are synthesized by the two-phase mini emulsion polymerization method and characterized by zeta-size analysis,Fourier transform infrared spectroscopy,and scanning electron microscopy.Afterwards,the patulin imprinted and the non-imprinted nanoparticles are attached on the surface of surface plasmon resonance(SPR)chips.The patulin imprinted and the non-imprinted SPR nanosensors are characterized by using atomic force microscope,ellipsometer,and contact angle measurements.Kinetic studies for patulin detection are carried out in the concentration range of 0.5nmol-750nmol.The limit of detection and the limit of quantification values are obtained as 0.011 nmol and 0.036 nmol,respectively.In all kinetic analysis,the response time is 13 min for equilibration,adsorption,and desorption cycles.The selectivity studies of the patulin imprinted and the non-imprinted SPR nanosensors are determined in the presence of ochratoxin A and aflatoxin Bl.In order to demonstrate the applicability,validation studies of the patulin imprinted SPR nanosensor are performed by liquid chromatography-tandem mass spectrometry(LC-MS).
基金funded by the Natural Sciences and Engineering Research Council of Canada(NSERC)by Canada Research Chairs Program.
文摘Patulin(PTL,154 Da)is a small molecule toxin produced as a mold metabolite.PTL is unstable in animals and is quickly converted to its metabolites.Thus,few successful antibodies were reported even after extensive efforts in hapten design.In this work,a library immobilization strategy was used to obtain DNA aptamers for PTL,where exposure of PTL to live animals or cells is no longer needed.After 20 rounds of selection,the most abundant aptamer named PTL-1 has a dissociation constant(K_(d))of 12.5μM based on isothermal titration calorimetry and 18.4μM based on thioflavin T(ThT)fluorescence spectroscopy.PTL-1 has excellent selectivity against nine other antibiotics.Compared to previously reported aptamers for PTL,PTL-1 is either shorter or has higher affinity.A label-free ThT fluorescence assay based on PTL-1 was developed,which has a detection limit of 0.2μM(detection range 0.2-20μM)for PTL in fruit juices.The PTL-1 aptamer shows resistance to matrix interference and can be applied to the rapid detection of PTL in a blended juice.This study is a nice demonstration of selecting aptamers for a target molecule difficult for the production of antibodies.
基金supported by the National Natural Science Foundation of China(Grant No.32301255 and 31772090)"Pioneer"and"Leading Goose"R&D Program of Zhejiang(Grants No.2024SSYS0103).
文摘Patulin(PAT)is a toxic fungal metabolite frequently detected in fruits and derived products,posing significant risks to food safety and public health.In this study,we identified and characterized an NADPH-dependent aldoketo reductase(LpAKR)from the probiotic strain Lactiplantibacillus plantarum CCFM1287 that efficiently catalyzes the conversion of PAT into the less toxic metabolite E-ascladiol.Functional disruption of the LpAKR gene in L.plantarum significantly reduced PAT-degrading capacity,confirming its essential role in the detoxification process.The recombinant LpAKR protein was expressed and purified,and its enzymatic activity toward PAT was confirmed in vitro.Structural modeling and molecular docking revealed that PAT binds in a hydrophobic pocket formed by residues Trp24,Trp116,Tyr53,Ala52,Leu78,and Met276,stabilized throughπ-πstacking with Trp116 and positioned adjacent to the NADPH nicotinamide ring.His111 and Asp48 were predicted to participate in proton and electron transfer during catalysis,and site-directed mutagenesis(His111D,Trp116A,Met276A)markedly decreased enzymatic activity,validating their catalytic importance.Molecular dynamics simulations further confirmed the role of these residues in substrate binding and structural stability.These findings provide mechanistic insight into PAT degradation by a food-grade aldo-keto reductase and highlight the potential application of L.plantarum CCFM1287 in enzymatic mycotoxin detoxification strategies for the food industry.
基金supported by the key projects for tackling key core technologies in Shaanxi Province(2024-NC-GJHX-12)the Natural Science Foundation of China(32172301 and 32202065).
文摘Selenium(Se)is an essential trace element for humans.Inorganic Se is converted into organic Se that is more easily absorbed during the growth of yeast.Patulin(PAT)is a fungal toxin that can cause damage to the intestinal mucosa.Se can alleviate intestinal damage,and this study mainly focuses on the research of Se enrichment in Kluyveromyces marxianus YG-4(SeK)and its repair effect on jejunal injury induced by PAT.In this study,K.marxianus YG-4 could enrich up to 721.05μg/g of total Se content,of which 86.30%was organic Se.In the organic Se of SeK,protein-bound Se,polysaccharide-bound Se,and nucleic acid-bound Se accounted for 83.22%,10.76%,and 2.68%,respectively.Moreover,SeK had an increased amino acid content compared with K.marxianus YG-4.Finally,the morphology and survival ability of K.marxianus YG-4 were not disrupted after Se enrichment,indicating that SeK has the potential to become a probiotic in the food industry.Furthermore,SeK had the ability to alleviate jejunal damage induced by PAT.Specifically,SeK alleviated intestinal tissue lesions and led to an increase in mucin levels.Moreover,SeK promoted the expression of tight junction proteins(claudin-1,occludin,and ZO-1)and inhibited the activation of the NF-κB signaling pathway.Additionally,SeK reduced harmful bacteria including Turicibacter,Anaerotruncus,and Desulfovibrio,and increased beneficial bacteria including Eggerthellaceae,Muribaculaceae,Dubosiella,and short chain fatty acid producing bacteria(Erysipelotrichaceae and Faecalibaculum).In summary,SeK is expected to become a dietary candidate for alleviating the damage caused by mycotoxins.
基金supported by the National Natural Science Foundation of China(32472804 and 32472414).
文摘Patulin(PAT)is a harmful mycotoxin that poses a serious threat to food safety,highlighting the urgent need for the development of efficient and environmentally friendly detoxification methods.This study identified a novel PAT-degrading enzyme,Glutaredoxin GRX3(GRX3)for the first time.Molecular docking results indicated that PAT stably binds to the hydrophobic cavity of GRX3.Key GRX3 residues interact with PAT through hydrogen bonds.Biochemical analyses revealed that GRX3 is highly active at moderate temperatures(28-45℃)and nearneutral pH and functions via glutathione(GSH)-dependent and thiol-mediated mechanisms.LC-MS/MS analysis confirmed that GRX3 catalyzed the PAT ring opening,converting it into E-ascladiol,which exhibited significantly reduced toxicity.In practical applications,GRX3 effectively degrades PAT in both freshly prepared and commercially available fruit juices,achieving nearly 100%degradation within 24 h in fresh juice.Overall,this study reveals a novel GSH-dependent detoxification pathway for PAT and establishes a strong mechanistic and practical application of GRX3 in agricultural products and food safety control.
