活性氧对白腐真菌漆酶合成的影响
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摘要
分别将不同活性氧在发酵第4天加入发酵产酶培养基中,一定时间后通过测生物量和漆酶酶活研究外界活性氧对白腐真菌漆酶合成的影响,并通过活性电泳进一步验证。结果表明:低浓度的活性氧可以通过信号传导方式调控漆酶合成,超氧阴离子抑制白腐真菌产漆酶,过氧化氢和羟自由基促进白腐真菌产漆酶。实验结果对漆酶合成调控、植物-病原菌相互作用以及生物防护具有重要意义。
In this paper,various of reactive oxygen species were respectively added to the fermentation medium on the 4 d. After a certain time,the effection of different reactive oxygen species on laccase synthesis of white-rot fungus,was evaluated by measuring biomass and laccase activity. And then this effection was further investigated by the method of native PAGE. The experiment results show that reactive oxygen species can regulate laccase synthesis of white-rot fungus via redox regulation in cellular signaling. In addition,superoxide anion inhibits laccase synthesis,but hydrogen peroxide and hydroxyl radical promote it obviously. These results are very important for researching biosynthesis regulation of laccase and interaction between plant and pathogen,which can be used for biological protection.
In this paper,various of reactive oxygen species were respectively added to the fermentation medium on the 4 d. After a certain time,the effection of different reactive oxygen species on laccase synthesis of white-rot fungus,was evaluated by measuring biomass and laccase activity. And then this effection was further investigated by the method of native PAGE. The experiment results show that reactive oxygen species can regulate laccase synthesis of white-rot fungus via redox regulation in cellular signaling. In addition,superoxide anion inhibits laccase synthesis,but hydrogen peroxide and hydroxyl radical promote it obviously. These results are very important for researching biosynthesis regulation of laccase and interaction between plant and pathogen,which can be used for biological protection.
引文
[1]黎劼.病原菌对植物的作用及其受环境因子的影响[J].河南科技,2013,12:223-225.
[2] WESENBERG D,KYRIAKIDES I,AGATHOS S N. White-rot fungi and their enzymes for the treatment of industrial dye effluents[J]. Biotechnology Advances,2003,22(1-2):161-187.
[3] ANTOSOVA Z,SYCHROVA H. Yeast hosts for the production of recombinant laccases:a review[J]. Molecular Biotechnology,2016,58(2):93-116.
[4] GALHAUP C,GOLLER S,PETERBAUER C K,et al. Characterization of the major laccase isoenzyme from Trametes pubescens and regulation of its synthesis by metal ions[J]. Microbiology-Sgm,2002,148:2159-2169.
[5] COLLINS P J,DOBSON A D W. Regulation of laccase gene transcription in Trametes versicolor[J]. Applied And Environmental Microbiology,1997,63(9):3444-3450.
[6] TERRON M C,GONZALEZ T,CARBAJO J M,et al. Structural close-related aromatic compounds have different effects on laccase activity and on lcc gene expression in the ligninolytic fungus Trametes sp I-62[J]. Fungal Genetics And Biology,2004,41(10):954-962.
[7] SUN Xiao,LIAO Xiangru,LIU Jiayang,et al. Laccase production under solid-state fermentation of rice straw by Pycnoporus sanguineus SYBC-L12 and its decolorization of synthetic dyes[J]. Journal of Food Science and Biotechnology,2013(3):240-249.(in Chinese)
[8] SODEN D M,DOBSON A D W. Differential regulation of laccase gene expression in Pleurotus sajor-caju[J]. MicrobiologySgm,2001,147:1755-1763.
[9] WANG Zhixin,CAI Yujie,LI Yanyan,et al. Effect of fermentation medium and conditions on laccase production by Pycnoporus sp. SYBC-L1[J]. Journal of Food Science and Biotechnology,2009(6):832-839.(in Chinese)
[10] WOJTASZEK P. Oxidative burst:an early plant response to pathogen infection[J]. Biochemical Journal,1997,322:681-692.
[11] WANG P F,KANG J H,ZHENG R L,et al. Scavenging effects of phenylpropanoid glycosides from Pedicularis on superoxide anion and hydroxyl radical by the spin trapping method(95)02255-4[J]. Biochemical Pharmacology,1996,51(5):687-691.
[12] JASZEK M,ZUCHOWSKI J,DAJCZAK E,et al. Ligninolytic enzymes can participate in a multiple response system to oxidative stress in white-rot basidiomycetes:Fomes fomentarius and Tyromyces pubescens[J]. International Biodeterioration&Biodegradation,2006,58(3-4):168-175.
[13] YANG Y,FAN F F,ZHUO R,et al. Expression of the laccase gene from a white rot fungus in Pichia pastoris can enhance the resistance of this yeast to H2O2-mediated oxidative stress by stimulating the glutathione-based antioxidative system[J]. Applied and Environmental Microbiology,2012,78(16):5845-5854.
[14] MAYER A M,STAPLES R C,GIL-AD N L. Mechanisms of survival of necrotrophic fungal plant pathogens in hosts expressing the hypersensitive response[J]. Phytochemistry,2001,58(1):33-41.
[15] LI H Y,WANG J S. Release of active oxygen from phytopathogenic bacteria and their regulation[J]. Chinese Science Bulletin,1999,44(1):71-75.
[16] ROLKE Y,LIU S J,QUIDDE T,et al. Functional analysis of H2O2-generating systems in Botrytis cinerea:the major Cu-Zn-superoxide dismutase(BCSOD1)contributes to virulence on French bean,whereas a glucose oxidase(BCGOD1)is dispensable[J]. Molecular Plant Pathology,2004,5(1):17-27.
[17] DEIGHTON N,MUCKENSCHNABEL I,GOODMAN B A,et al. Lipid peroxidation and the oxidative burst associated with infection of Capsicum annuum by Botrytis cinerea[J]. Plant Journal,1999,20(4):485-492.
[18] TORRES M A. ROS in biotic interactions[J]. Physiologia Plantarum,2010,138(4):414-429.
[19] RAY P D,HUANG B W,TSUJI Y. Reactive oxygen species(ROS)homeostasis and redox regulation in cellular signaling[J].Cell Signal,2012,24(5):981-990.
[20] LIU Jiayang,CAI Yujie,LIAO Xiangru,et al.Screening and optimization of laccase production froma laccase producing fungal[J].Food&Machinery,2010(4):10-14.(in Chinese)
[21]刘家扬.白腐真菌Pycnoporus sp. SYBC-L3产漆酶及应用研究[D].无锡:江南大学,2013.
[22] WANG Z X,CAI Y J,LIAO X R,et al. Production and characterization of a novel laccase with cold adaptation and high thermal stability from an isolated fungus[J]. Applied Biochemistry And Biotechnology,2010,162(1):280-294.
[23] TEERAPATSAKUL C,PARRA R,BUCKE C,et al. Improvement of laccase production from Ganoderma sp KU-Alk4 by medium engineering[J]. World Journal of Microbiology&Biotechnology,2007,23(11):1519-1527.
[24] ZENG X K,CAI Y J,LIAO X R,et al. Decolorization of synthetic dyes by crude laccase from a newly isolated Trametes trogii strain cultivated on solid agro-industrial residue[J]. Journal of Hazardous Materials,2011,187(1-3):517-525.
[25] BELINKY P A,FLIKSHTEIN N,LECHENKO S,et al. Reactive oxygen species and induction of lignin peroxidase in Phanerochaete chrysosporium[J]. Applied And Environmental Microbiology,2003,69(11):6500-6506.
[26] GEORGIOU C D,PETROPOULOU K P. Effect of the antioxidant ascorbic acid on sclerotial differentiation in Rhizoctonia solani[J]. Plant Pathology,2001,50(5):594-600.
[27] SPICKETT C M,SMIRNOFF N,PITT A R. The biosynthesis of erythroascorbate in Saccharomyces cerevisiae and its role as an antioxidant[J]. Free Radical Biology And Medicine,2000,28(2):183-192.
[28] LIU J Y,CAI Y J,LIAO X R,et al. Efficiency of laccase production in a 65-L air-lift reactor for potential green industrial and environmental application[J]. Journal of Cleaner Production,2013,39:154-160.
[29] BIAN Dan,QIU Luhui,YANG Hailong. Effects of diverse metal ions and pH on laccase production of Ganoderma Lucidum[J].Journal of Wenzhou University(Natural Sciences),2015(2):58-62.(in Chinese)
[30] CHEN Daidi,NIU Jiezhen,YU Xiaoyuan,et al.The factors affecting the expression and activity of laccase in fungi[J]. Chinese Bulletin of Life Sciences,2013,11:1053-1058.(in Chinese)
[31] CHEN S X,SCHOPFER P. Hydroxyl-radical production in physiological reactions-a novel function of peroxidase[J]. European Journal of Biochemistry,1999,260(3):726-735.
[32] BISHOP D F,STERN G,FLEISCHMAN M,et al. Hydrogen peroxide catalytic oxidation of refractory organics in municipal waste waters[J]. Industrial&Engineering Chemistry Process Design and Development,1968,7:110-117.
[2] WESENBERG D,KYRIAKIDES I,AGATHOS S N. White-rot fungi and their enzymes for the treatment of industrial dye effluents[J]. Biotechnology Advances,2003,22(1-2):161-187.
[3] ANTOSOVA Z,SYCHROVA H. Yeast hosts for the production of recombinant laccases:a review[J]. Molecular Biotechnology,2016,58(2):93-116.
[4] GALHAUP C,GOLLER S,PETERBAUER C K,et al. Characterization of the major laccase isoenzyme from Trametes pubescens and regulation of its synthesis by metal ions[J]. Microbiology-Sgm,2002,148:2159-2169.
[5] COLLINS P J,DOBSON A D W. Regulation of laccase gene transcription in Trametes versicolor[J]. Applied And Environmental Microbiology,1997,63(9):3444-3450.
[6] TERRON M C,GONZALEZ T,CARBAJO J M,et al. Structural close-related aromatic compounds have different effects on laccase activity and on lcc gene expression in the ligninolytic fungus Trametes sp I-62[J]. Fungal Genetics And Biology,2004,41(10):954-962.
[7] SUN Xiao,LIAO Xiangru,LIU Jiayang,et al. Laccase production under solid-state fermentation of rice straw by Pycnoporus sanguineus SYBC-L12 and its decolorization of synthetic dyes[J]. Journal of Food Science and Biotechnology,2013(3):240-249.(in Chinese)
[8] SODEN D M,DOBSON A D W. Differential regulation of laccase gene expression in Pleurotus sajor-caju[J]. MicrobiologySgm,2001,147:1755-1763.
[9] WANG Zhixin,CAI Yujie,LI Yanyan,et al. Effect of fermentation medium and conditions on laccase production by Pycnoporus sp. SYBC-L1[J]. Journal of Food Science and Biotechnology,2009(6):832-839.(in Chinese)
[10] WOJTASZEK P. Oxidative burst:an early plant response to pathogen infection[J]. Biochemical Journal,1997,322:681-692.
[11] WANG P F,KANG J H,ZHENG R L,et al. Scavenging effects of phenylpropanoid glycosides from Pedicularis on superoxide anion and hydroxyl radical by the spin trapping method(95)02255-4[J]. Biochemical Pharmacology,1996,51(5):687-691.
[12] JASZEK M,ZUCHOWSKI J,DAJCZAK E,et al. Ligninolytic enzymes can participate in a multiple response system to oxidative stress in white-rot basidiomycetes:Fomes fomentarius and Tyromyces pubescens[J]. International Biodeterioration&Biodegradation,2006,58(3-4):168-175.
[13] YANG Y,FAN F F,ZHUO R,et al. Expression of the laccase gene from a white rot fungus in Pichia pastoris can enhance the resistance of this yeast to H2O2-mediated oxidative stress by stimulating the glutathione-based antioxidative system[J]. Applied and Environmental Microbiology,2012,78(16):5845-5854.
[14] MAYER A M,STAPLES R C,GIL-AD N L. Mechanisms of survival of necrotrophic fungal plant pathogens in hosts expressing the hypersensitive response[J]. Phytochemistry,2001,58(1):33-41.
[15] LI H Y,WANG J S. Release of active oxygen from phytopathogenic bacteria and their regulation[J]. Chinese Science Bulletin,1999,44(1):71-75.
[16] ROLKE Y,LIU S J,QUIDDE T,et al. Functional analysis of H2O2-generating systems in Botrytis cinerea:the major Cu-Zn-superoxide dismutase(BCSOD1)contributes to virulence on French bean,whereas a glucose oxidase(BCGOD1)is dispensable[J]. Molecular Plant Pathology,2004,5(1):17-27.
[17] DEIGHTON N,MUCKENSCHNABEL I,GOODMAN B A,et al. Lipid peroxidation and the oxidative burst associated with infection of Capsicum annuum by Botrytis cinerea[J]. Plant Journal,1999,20(4):485-492.
[18] TORRES M A. ROS in biotic interactions[J]. Physiologia Plantarum,2010,138(4):414-429.
[19] RAY P D,HUANG B W,TSUJI Y. Reactive oxygen species(ROS)homeostasis and redox regulation in cellular signaling[J].Cell Signal,2012,24(5):981-990.
[20] LIU Jiayang,CAI Yujie,LIAO Xiangru,et al.Screening and optimization of laccase production froma laccase producing fungal[J].Food&Machinery,2010(4):10-14.(in Chinese)
[21]刘家扬.白腐真菌Pycnoporus sp. SYBC-L3产漆酶及应用研究[D].无锡:江南大学,2013.
[22] WANG Z X,CAI Y J,LIAO X R,et al. Production and characterization of a novel laccase with cold adaptation and high thermal stability from an isolated fungus[J]. Applied Biochemistry And Biotechnology,2010,162(1):280-294.
[23] TEERAPATSAKUL C,PARRA R,BUCKE C,et al. Improvement of laccase production from Ganoderma sp KU-Alk4 by medium engineering[J]. World Journal of Microbiology&Biotechnology,2007,23(11):1519-1527.
[24] ZENG X K,CAI Y J,LIAO X R,et al. Decolorization of synthetic dyes by crude laccase from a newly isolated Trametes trogii strain cultivated on solid agro-industrial residue[J]. Journal of Hazardous Materials,2011,187(1-3):517-525.
[25] BELINKY P A,FLIKSHTEIN N,LECHENKO S,et al. Reactive oxygen species and induction of lignin peroxidase in Phanerochaete chrysosporium[J]. Applied And Environmental Microbiology,2003,69(11):6500-6506.
[26] GEORGIOU C D,PETROPOULOU K P. Effect of the antioxidant ascorbic acid on sclerotial differentiation in Rhizoctonia solani[J]. Plant Pathology,2001,50(5):594-600.
[27] SPICKETT C M,SMIRNOFF N,PITT A R. The biosynthesis of erythroascorbate in Saccharomyces cerevisiae and its role as an antioxidant[J]. Free Radical Biology And Medicine,2000,28(2):183-192.
[28] LIU J Y,CAI Y J,LIAO X R,et al. Efficiency of laccase production in a 65-L air-lift reactor for potential green industrial and environmental application[J]. Journal of Cleaner Production,2013,39:154-160.
[29] BIAN Dan,QIU Luhui,YANG Hailong. Effects of diverse metal ions and pH on laccase production of Ganoderma Lucidum[J].Journal of Wenzhou University(Natural Sciences),2015(2):58-62.(in Chinese)
[30] CHEN Daidi,NIU Jiezhen,YU Xiaoyuan,et al.The factors affecting the expression and activity of laccase in fungi[J]. Chinese Bulletin of Life Sciences,2013,11:1053-1058.(in Chinese)
[31] CHEN S X,SCHOPFER P. Hydroxyl-radical production in physiological reactions-a novel function of peroxidase[J]. European Journal of Biochemistry,1999,260(3):726-735.
[32] BISHOP D F,STERN G,FLEISCHMAN M,et al. Hydrogen peroxide catalytic oxidation of refractory organics in municipal waste waters[J]. Industrial&Engineering Chemistry Process Design and Development,1968,7:110-117.