拟粉红锁掷孢酵母降解展青霉素的蛋白质组学
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摘要
展青霉素(PAT)又称棒曲霉素,是一种主要由曲霉属、青霉属等产生的有毒的真菌次级代谢产物,主要污染水果及其制品,具有细胞毒性、致畸性及遗传毒性等,严重威胁人类的健康。近年来,生物防治因其安全、绿色、高效等优点成为一种具有良好发展前景的控制水果真菌毒素产生的方法。有研究表明,部分酵母菌可以直接抑制PAT的产生,有的甚至可以降解PAT,但具体的降解机制尚不明确。作者所在实验室前期研究发现,拟粉红锁掷孢酵母(Sporidiobolus pararoseus Y16)能够在体外降解PAT,但具体的降解机制尚未明确,作者利用蛋白质组学技术对S. pararoseus Y16降解PAT的机制进行了初步研究,可以为真菌毒素的生物降解提供新的理论依据。
Patulin(PAT) is one of toxic fungal secondary metabolites,mainly produced by Aspergillus sp and Penicillium sp,it has been identified in various fruits and fruit products all over the world,and it has cytotoxicity,teratogenicity and genotoxicity,hence,it poses potential hazard to human and animals. In recent years,biological control has become a promising method to control the mycotoxin produced by fruits because of its advantages of safety,green and high efficiency. It has been reported that,some strains of yeast can directly inhibit the production of PAT,and even degrade PAT,however,the mechanism of controlling and degradation PAT underlying is not clear. Our previous study found that,Sporidiobolus pararoseus Y16 can significantly degrade PAT in vitro,but the possible mechanism is not clear,in this study,the mechanism of degradation of PAT by S.pararoseus using proteomic technology was investigated. This study can provide a new theoretical evidence for the biological degradation of mycotoxins.
Patulin(PAT) is one of toxic fungal secondary metabolites,mainly produced by Aspergillus sp and Penicillium sp,it has been identified in various fruits and fruit products all over the world,and it has cytotoxicity,teratogenicity and genotoxicity,hence,it poses potential hazard to human and animals. In recent years,biological control has become a promising method to control the mycotoxin produced by fruits because of its advantages of safety,green and high efficiency. It has been reported that,some strains of yeast can directly inhibit the production of PAT,and even degrade PAT,however,the mechanism of controlling and degradation PAT underlying is not clear. Our previous study found that,Sporidiobolus pararoseus Y16 can significantly degrade PAT in vitro,but the possible mechanism is not clear,in this study,the mechanism of degradation of PAT by S.pararoseus using proteomic technology was investigated. This study can provide a new theoretical evidence for the biological degradation of mycotoxins.
引文
[1]MARIN S,RAMOS A J,CANO-SANCHO G,et al.Mycotoxins:occurrence,toxicology,and exposure assessment[J].Food&Chemical Toxicology An International Journal Published for the British Industrial Biological Research Association,2013,60(10):218-237.
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[3]WU T S,LIAO Y C,YU F Y,et al.Mechanism of patulin-induced apoptosis in human leukemia cells(HL-60)[J].Toxicology Letters,2008,183(3):105-111.
[4]DONMEZALTUNTAS H,GOKALPYILDIZ P,BITGEN N,et al.Evaluation of genotoxicity,cytotoxicity and cytostasis in human lymphocytes exposed to patulin by using the cytokinesis-block micronucleus cytome(CBMN cyt)assay[J].Mycotoxin Research,2013,29(2):63.
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[7]PUEL O,GALTIER P,OSWALD I P.Biosynthesis and toxicological effects of patulin[J].Toxins,2010,2(4):613.
[8]EGMOND H P V,JONKER M A.Chapter 3-regulations and limits for mycotoxins in fruits and vegetables[J].Mycotoxins in Fruits&Vegetables,2008:45-74.
[9]ZHANG H.Analysis of《the limited of mycotoxins in food》new standard[J].Food Safety Guide,2013(1):46-47.
[10]杨其亚.植酸增强胶红酵母对草莓、苹果及桃果采后病害的控制效应[D].镇江:江苏大学,2011.
[11]YANG Q,ZHANG H,ZHANG X,et al.Phytic acid enhances biocontrol activity of Rhodotorula mucilaginosa against Penicillium expansum contamination and patulin production in apples[J].Frontiers in Microbiology,2015,6(8):1296.
[12]CAO J,ZHANG H,YANG Q,et al.Efficacy of Pichia caribbica in controlling blue mold rot and patulin degradation in apples[J].International Journal of Food Microbiology,2013,162(2):167.
[13]CASTORIA R,MANNINA L,DUR魣NPATR譫N R,et al.Conversion of the mycotoxin patulin to the less toxic desoxypatulinic acid by the biocontrol yeast Rhodosporidium kratochvilovae strain LS11[J].Journal of Agricultural&Food Chemistry,2011,59(21):11571-11578.
[14]CASTORIA R,MORENA V,CAPUTO L,et al.Effect of the biocontrol yeast Rhodotorula glutinis strain LS11 on patulin accumulation in stored apples[J].Phytopathology,2005,95(11):1271-1278.
[15]ZHU R,YU T,GUO S,et al.Effect of the yeast Rhodosporidium paludigenum on postharvest decay and patulin accumulation in apples and pears[J].Journal of Food Protection,2015,78(1):157-163.
[16]LI Q,LI C,LI P,et al.The biocontrol effect of Sporidiobolus pararoseus Y16 against postharvest diseases in table grapes caused by Aspergillus niger,and the possible mechanisms involved[J].Biological Control,2017,113:18-25.
[17]AKUTSU J I,ZHANG Z,MORITA R,et al.Identification and characterization of a thermostable bifunctional enzyme with phosphomannose isomerase and sugar-1-phosphate nucleotidylyltransferase activities from a hyperthermophilic archaeon,Pyrococcus horikoshii OT3[J].Extremophiles Life Under Extreme Conditions,2015,19(6):1077-1085.
[18]BRADFORD M M.A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding[J].Analytical Biochemistry,1976,72(s1-2):248-254.
[19]CHOI W I,JEON B N,YUN C O,et al.Proto-oncogene FBI-1 represses transcription of p21CIP1 by inhibition of transcription activation by p53 and Sp1[J].Journal of Biological Chemistry,2009,284(19):12633-12644.
[20]XU Danning,TIAN Yunbo,HUANG Yumao,et al.Advances in protective effect of HSP70/HSP27 on immune cells[J].Journal of Zhongkai University of Agriculture and Engineering,2011,24(1):56-60.(in Chinese)
[21]NEUER A,MELE C,LIU H C,et al.Monoclonal antibodies to mammalian heat shock proteins impair mouse embryo development in vitro[J].Human Reproduction,1998,13(4):987-990.
[22]CAI B,LI Q,XU Y,et al.Genome-wide analysis of the fructose 1,6-bisphosphate aldolase(FBA)gene family and functional characterization of FBA7 in tomato[J].Plant Physiology&Biochemistry,2016,108:251-265.
[2]SCHUMACHER D M,METZLER M,LEHMANN L.Mutagenicity of the mycotoxin patulin in cultured Chinese hamster V79cells,and its modulation by intracellular glutathione[J].Archives of Toxicology,2005,79(2):110-21.
[3]WU T S,LIAO Y C,YU F Y,et al.Mechanism of patulin-induced apoptosis in human leukemia cells(HL-60)[J].Toxicology Letters,2008,183(3):105-111.
[4]DONMEZALTUNTAS H,GOKALPYILDIZ P,BITGEN N,et al.Evaluation of genotoxicity,cytotoxicity and cytostasis in human lymphocytes exposed to patulin by using the cytokinesis-block micronucleus cytome(CBMN cyt)assay[J].Mycotoxin Research,2013,29(2):63.
[5]周思敏.棒曲霉毒素对HepG2细胞的遗传毒性及氧化应激机制的研究[D].大连:大连医科大学,2009.
[6]WRIGHT S A I,FELICE D V D,IANIRI G,et al.Two rapid assays for screening of patulin biodegradation[J].International Journal of Environmental Science&Technology,2014,11(5):1387-1398.
[7]PUEL O,GALTIER P,OSWALD I P.Biosynthesis and toxicological effects of patulin[J].Toxins,2010,2(4):613.
[8]EGMOND H P V,JONKER M A.Chapter 3-regulations and limits for mycotoxins in fruits and vegetables[J].Mycotoxins in Fruits&Vegetables,2008:45-74.
[9]ZHANG H.Analysis of《the limited of mycotoxins in food》new standard[J].Food Safety Guide,2013(1):46-47.
[10]杨其亚.植酸增强胶红酵母对草莓、苹果及桃果采后病害的控制效应[D].镇江:江苏大学,2011.
[11]YANG Q,ZHANG H,ZHANG X,et al.Phytic acid enhances biocontrol activity of Rhodotorula mucilaginosa against Penicillium expansum contamination and patulin production in apples[J].Frontiers in Microbiology,2015,6(8):1296.
[12]CAO J,ZHANG H,YANG Q,et al.Efficacy of Pichia caribbica in controlling blue mold rot and patulin degradation in apples[J].International Journal of Food Microbiology,2013,162(2):167.
[13]CASTORIA R,MANNINA L,DUR魣NPATR譫N R,et al.Conversion of the mycotoxin patulin to the less toxic desoxypatulinic acid by the biocontrol yeast Rhodosporidium kratochvilovae strain LS11[J].Journal of Agricultural&Food Chemistry,2011,59(21):11571-11578.
[14]CASTORIA R,MORENA V,CAPUTO L,et al.Effect of the biocontrol yeast Rhodotorula glutinis strain LS11 on patulin accumulation in stored apples[J].Phytopathology,2005,95(11):1271-1278.
[15]ZHU R,YU T,GUO S,et al.Effect of the yeast Rhodosporidium paludigenum on postharvest decay and patulin accumulation in apples and pears[J].Journal of Food Protection,2015,78(1):157-163.
[16]LI Q,LI C,LI P,et al.The biocontrol effect of Sporidiobolus pararoseus Y16 against postharvest diseases in table grapes caused by Aspergillus niger,and the possible mechanisms involved[J].Biological Control,2017,113:18-25.
[17]AKUTSU J I,ZHANG Z,MORITA R,et al.Identification and characterization of a thermostable bifunctional enzyme with phosphomannose isomerase and sugar-1-phosphate nucleotidylyltransferase activities from a hyperthermophilic archaeon,Pyrococcus horikoshii OT3[J].Extremophiles Life Under Extreme Conditions,2015,19(6):1077-1085.
[18]BRADFORD M M.A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding[J].Analytical Biochemistry,1976,72(s1-2):248-254.
[19]CHOI W I,JEON B N,YUN C O,et al.Proto-oncogene FBI-1 represses transcription of p21CIP1 by inhibition of transcription activation by p53 and Sp1[J].Journal of Biological Chemistry,2009,284(19):12633-12644.
[20]XU Danning,TIAN Yunbo,HUANG Yumao,et al.Advances in protective effect of HSP70/HSP27 on immune cells[J].Journal of Zhongkai University of Agriculture and Engineering,2011,24(1):56-60.(in Chinese)
[21]NEUER A,MELE C,LIU H C,et al.Monoclonal antibodies to mammalian heat shock proteins impair mouse embryo development in vitro[J].Human Reproduction,1998,13(4):987-990.
[22]CAI B,LI Q,XU Y,et al.Genome-wide analysis of the fructose 1,6-bisphosphate aldolase(FBA)gene family and functional characterization of FBA7 in tomato[J].Plant Physiology&Biochemistry,2016,108:251-265.