A new fungal peroxidase with alkaline-tolerant,chloride-enhancing activity and dye decolorization capacity
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- 英文论文题名：A new fungal peroxidase with alkaline-tolerant,chloride-enhancing activity and dye decolorization capacity
- 论文作者：Jing Si ; Bao-Kai Cui ; Yu-Cheng Dai
- 英文论文作者：Jing Si ; Bao-Kai Cui ; Yu-Cheng Dai ; Institute of Microbiology ; Beijing Forestry University
- 年：2016
- 作者机构：Institute of Microbiology,Beijing Forestry University;
- 英文论文关键词：Perenniporia subacida peroxidase ; purification ; alkaline tolerance ; chloride-enhancing activity ; dye decolorization capacity
- 会议召开时间：2016-08-19
- 会议录名称：中国菌物学会2016年学术年会论文摘要集
- 英文会议录名称：Abstracts of 2016 Mycological Society of China Annual Meeting
- 语种：英文
- 分类号：Q939.9
- 学会代码：ZGVL
- 会议名称：中国菌物学会2016年学术年会
- 会议地点：中国福建福州
- 主办单位：中国菌物学会
- 学会名称：中国菌物学会
- 页数：3
- 文件大小：725k
- 原文格式：D
- 会议级别：全国
摘要
A new fungal peroxidase(Pspd) from Perenniporia subacida was purified by ammonium sulfate precipitation,DEAE-cellulose DE52 anionic exchange and Sepharose CL-6B chromatography,resulting in a high specific activity of 9.138 U/mg,3.622-fold higher than that of crude enzyme at the same level.Polyacrylamide gel electrophoresis and UV-visible adsorption spectrum analysis showed that the purified enzyme is a heme-containing monomer with a molecular mass of 43.0 k Da.Optimal peroxidase activity was obtained at p H 5.5 and 30°C when using 100.0 m M n-propanol as substrate,and under these conditions,the catalytic efficiency(kcat/Km) is 1.57 s-1 μM-1.Pspd was inhibited by L-cysteine,dithiothreitol,EDTA and sodium azide,but stimulated by Mn2+,Na+,Mg2+ and K+.The enzyme is stable over a broad p H range of 7.0-8.5 after incubation for 72 h,which indicated that the enzyme is lasting alkaline-tolerant.It was worth noting that the chloride at relatively low concentrations can enhance the peroxidase activity,with concomitant increase in substrate affinity.Additionally,Pspd performed high decolorization capability toward structurally various dyes and the capability was independent of the oxidizing mediators,with 75.31% of Neutral Red(50.0 mg/L) being decolorized by 1.5 U/m L pure enzyme after incubation for 72 h.These properties demonstrated that Pspd has potentials for textile dyes decolorization applications.
A new fungal peroxidase(Pspd) from Perenniporia subacida was purified by ammonium sulfate precipitation,DEAE-cellulose DE52 anionic exchange and Sepharose CL-6B chromatography,resulting in a high specific activity of 9.138 U/mg,3.622-fold higher than that of crude enzyme at the same level.Polyacrylamide gel electrophoresis and UV-visible adsorption spectrum analysis showed that the purified enzyme is a heme-containing monomer with a molecular mass of 43.0 k Da.Optimal peroxidase activity was obtained at p H 5.5 and 30°C when using 100.0 m M n-propanol as substrate,and under these conditions,the catalytic efficiency(kcat/Km) is 1.57 s-1 μM-1.Pspd was inhibited by L-cysteine,dithiothreitol,EDTA and sodium azide,but stimulated by Mn2+,Na+,Mg2+ and K+.The enzyme is stable over a broad p H range of 7.0-8.5 after incubation for 72 h,which indicated that the enzyme is lasting alkaline-tolerant.It was worth noting that the chloride at relatively low concentrations can enhance the peroxidase activity,with concomitant increase in substrate affinity.Additionally,Pspd performed high decolorization capability toward structurally various dyes and the capability was independent of the oxidizing mediators,with 75.31% of Neutral Red(50.0 mg/L) being decolorized by 1.5 U/m L pure enzyme after incubation for 72 h.These properties demonstrated that Pspd has potentials for textile dyes decolorization applications.
A new fungal peroxidase(Pspd) from Perenniporia subacida was purified by ammonium sulfate precipitation,DEAE-cellulose DE52 anionic exchange and Sepharose CL-6B chromatography,resulting in a high specific activity of 9.138 U/mg,3.622-fold higher than that of crude enzyme at the same level.Polyacrylamide gel electrophoresis and UV-visible adsorption spectrum analysis showed that the purified enzyme is a heme-containing monomer with a molecular mass of 43.0 k Da.Optimal peroxidase activity was obtained at p H 5.5 and 30°C when using 100.0 m M n-propanol as substrate,and under these conditions,the catalytic efficiency(kcat/Km) is 1.57 s-1 μM-1.Pspd was inhibited by L-cysteine,dithiothreitol,EDTA and sodium azide,but stimulated by Mn2+,Na+,Mg2+ and K+.The enzyme is stable over a broad p H range of 7.0-8.5 after incubation for 72 h,which indicated that the enzyme is lasting alkaline-tolerant.It was worth noting that the chloride at relatively low concentrations can enhance the peroxidase activity,with concomitant increase in substrate affinity.Additionally,Pspd performed high decolorization capability toward structurally various dyes and the capability was independent of the oxidizing mediators,with 75.31% of Neutral Red(50.0 mg/L) being decolorized by 1.5 U/m L pure enzyme after incubation for 72 h.These properties demonstrated that Pspd has potentials for textile dyes decolorization applications.
引文
Bradford MM,1976.A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding.Analytical Biochemistry,72:248-254.
Eisenman HC,Mues M,Weber SE,Frases S,Chaskes S,Gerfen G,Casadevall A,2007.Cryptococcus neoformans laccase catalyses melanin synthesis from both D-and L-DOPA.Microbiology,153:3954-3962.
Enaud E,Trovaslet M,Naveau F,Decristoforo A,Bizet S,Vanhulle S,Jolivalt C,2011.Laccase chloride inhibition reduction by an anthraquinonic substrate.Enzyme and Microbial Technology,49:517-525.
Enayatizamir N,Tabandeh F,Rodríguez-Couto S,Yakhchali B,Alikhani HA,Mohammadi L,2011.
Biodegradation pathway and detoxification of the diazo dye Reactive Black 5 by Phanerochaete chrysosporium.Bioresource Technology,102:10359-10362.
Fang ZM,Li TL,Chang F,Zhou P,Fang W,Hong YZ,Zhang XC,Peng H,Xiao YZ,2012.A new marine bacterial laccase with chloride-enhancing,alkaline-dependent activity and dye decolorization ability.Bioresource Technology,111:36-41.
Fodil D,Jaouadi B,Badis A,Nadia ZJ,Ferradji FZ,Bejar S,Boutoumi H,2012.A thermostable humic acid peroxidase from Streptomyces sp.strain AH4:purification and biochemical characterization.Bioresource Technology,111:383-390.
Gopinath KP,Kathiravan MN,Srinivasan R,Sankaranarayanan S,2011.Evaluation and elimination of inhibitory effects of salts and heavy metal ions on biodegradation of Congo red by Pseudomonas sp.mutant.Bioresource Technology,102:3687-3693.
Jadhav JP,Kalyani DC,Telke AA,Phugare SS,Govindwar SP,2010.Evaluation of the efficacy of a bacterial consortium for the removal of color,reduction of heavy metals,and toxicity from textile dye effluent.Bioresource Technology,101:165-173.
Kalpana D,Shim JH,Oh BT,Senthil K,Lee YS,2011.Bioremediation of the heavy metal complex dye Isolan Dark Blue 2SGL-01 by white rot fungus Irpex lacteus.Journal of Hazardous Materials,198:198-205.
Kalyani DC,Phugare SS,Shedbalkar UU,Jadhav JP,2011.Purification and characterization of a bacterial peroxidase from the isolated strain Pseudomonas sp.SUK1 and its application for textile dye decolorization.Annals of Microbiology,61:483-491.
Kittl R,Mueangtoom K,Gonaus C,Khazaneh ST,Sygmund C,Haltrich D,Ludwig R,2012.A chloride tolerant laccase from the plant pathogen ascomycete Botrytis aclada expressed at high levels in Pichia pastoris.Journal of Biotechnology,157:304-314.
Niladevi KN,Jacob N,Prema P,2008.Evidence for a halotolerant-alkaline laccase in Streptomyces psammoticus:purification and characterization.Process Biochemistry,43:654-660.
Ntougias S,Baldrian P,Ehaliotis C,Nerud F,Antoniou T,MerhautováV,Zervakis GI,2012.Biodegradation and detoxification of olive mill wastewater by selected strains of the mushroom genera Ganoderma and Pleurotus.Chemosphere,88:620-626.
Paszczyński A,Huynh VB,Crawford R,1986.Comparison of ligninase-I and peroxidase-M2 from the white-rot fungus Phanerochaete chrysosporium.Archives of Biochemistry and Biophysics,244:750-765.
Patil P,Desai N,Govindwar S,Jadhav JP,Bapat V,2009.Degradation analysis of Reactive Red 198 by hairy roots of Tagetes patula L.(Marigold).Planta,230:725-735.
Renganathan V,Miki K,Gold MH,1985.Multiple molecular forms of diarylpropane oxygenase,an H2O2-requiring,lignin-degrading enzyme from Phanerochaete chrysosporium.Archives of Biochemistry and Biophysics,241:304-314.
Shanmugam V,Kumari M,Yadav KD,1999.n-Propanol as a substrate for assaying the ligninperoxidase activity of Phanerochaete chrysosporium.Indian Journal of Biochemistry and Biophysics,36:39-43.
Sutay Kocabas D,Bakir U,Phillips SE,Mc Pherson MJ,Ogel ZB,2008.Purification,characterization,and identification of a novel bifunctional catalase-phenol oxidase from Scytalidium thermophilum.Applied Microbiology and Biotechnology,79:407-415.
Thakkar AP,Dhamankar VS,Kapadnis BP,2006.Biocatalytic decolourisation of molasses by Phanerochaete chrysosporium.Bioresource Technology,97:1377-1381.
Tzika ED,Sotiroudis TG,Papadimitriou V,Xenakis A,2009.Partial purification and characterization of peroxidase from olives(Olea europaea cv.Koroneiki).European Food Research and Technology,228:487-495.
Yu GC,Wen XH,Li R,Qian Y,2006.In vitro degradation of a reactive azo dye by crude ligninolytic enzymes from nonimmersed liquid culture of Phanerochaete chrysosporium.Process Biochemistry,41:1987-1993.
Eisenman HC,Mues M,Weber SE,Frases S,Chaskes S,Gerfen G,Casadevall A,2007.Cryptococcus neoformans laccase catalyses melanin synthesis from both D-and L-DOPA.Microbiology,153:3954-3962.
Enaud E,Trovaslet M,Naveau F,Decristoforo A,Bizet S,Vanhulle S,Jolivalt C,2011.Laccase chloride inhibition reduction by an anthraquinonic substrate.Enzyme and Microbial Technology,49:517-525.
Enayatizamir N,Tabandeh F,Rodríguez-Couto S,Yakhchali B,Alikhani HA,Mohammadi L,2011.
Biodegradation pathway and detoxification of the diazo dye Reactive Black 5 by Phanerochaete chrysosporium.Bioresource Technology,102:10359-10362.
Fang ZM,Li TL,Chang F,Zhou P,Fang W,Hong YZ,Zhang XC,Peng H,Xiao YZ,2012.A new marine bacterial laccase with chloride-enhancing,alkaline-dependent activity and dye decolorization ability.Bioresource Technology,111:36-41.
Fodil D,Jaouadi B,Badis A,Nadia ZJ,Ferradji FZ,Bejar S,Boutoumi H,2012.A thermostable humic acid peroxidase from Streptomyces sp.strain AH4:purification and biochemical characterization.Bioresource Technology,111:383-390.
Gopinath KP,Kathiravan MN,Srinivasan R,Sankaranarayanan S,2011.Evaluation and elimination of inhibitory effects of salts and heavy metal ions on biodegradation of Congo red by Pseudomonas sp.mutant.Bioresource Technology,102:3687-3693.
Jadhav JP,Kalyani DC,Telke AA,Phugare SS,Govindwar SP,2010.Evaluation of the efficacy of a bacterial consortium for the removal of color,reduction of heavy metals,and toxicity from textile dye effluent.Bioresource Technology,101:165-173.
Kalpana D,Shim JH,Oh BT,Senthil K,Lee YS,2011.Bioremediation of the heavy metal complex dye Isolan Dark Blue 2SGL-01 by white rot fungus Irpex lacteus.Journal of Hazardous Materials,198:198-205.
Kalyani DC,Phugare SS,Shedbalkar UU,Jadhav JP,2011.Purification and characterization of a bacterial peroxidase from the isolated strain Pseudomonas sp.SUK1 and its application for textile dye decolorization.Annals of Microbiology,61:483-491.
Kittl R,Mueangtoom K,Gonaus C,Khazaneh ST,Sygmund C,Haltrich D,Ludwig R,2012.A chloride tolerant laccase from the plant pathogen ascomycete Botrytis aclada expressed at high levels in Pichia pastoris.Journal of Biotechnology,157:304-314.
Niladevi KN,Jacob N,Prema P,2008.Evidence for a halotolerant-alkaline laccase in Streptomyces psammoticus:purification and characterization.Process Biochemistry,43:654-660.
Ntougias S,Baldrian P,Ehaliotis C,Nerud F,Antoniou T,MerhautováV,Zervakis GI,2012.Biodegradation and detoxification of olive mill wastewater by selected strains of the mushroom genera Ganoderma and Pleurotus.Chemosphere,88:620-626.
Paszczyński A,Huynh VB,Crawford R,1986.Comparison of ligninase-I and peroxidase-M2 from the white-rot fungus Phanerochaete chrysosporium.Archives of Biochemistry and Biophysics,244:750-765.
Patil P,Desai N,Govindwar S,Jadhav JP,Bapat V,2009.Degradation analysis of Reactive Red 198 by hairy roots of Tagetes patula L.(Marigold).Planta,230:725-735.
Renganathan V,Miki K,Gold MH,1985.Multiple molecular forms of diarylpropane oxygenase,an H2O2-requiring,lignin-degrading enzyme from Phanerochaete chrysosporium.Archives of Biochemistry and Biophysics,241:304-314.
Shanmugam V,Kumari M,Yadav KD,1999.n-Propanol as a substrate for assaying the ligninperoxidase activity of Phanerochaete chrysosporium.Indian Journal of Biochemistry and Biophysics,36:39-43.
Sutay Kocabas D,Bakir U,Phillips SE,Mc Pherson MJ,Ogel ZB,2008.Purification,characterization,and identification of a novel bifunctional catalase-phenol oxidase from Scytalidium thermophilum.Applied Microbiology and Biotechnology,79:407-415.
Thakkar AP,Dhamankar VS,Kapadnis BP,2006.Biocatalytic decolourisation of molasses by Phanerochaete chrysosporium.Bioresource Technology,97:1377-1381.
Tzika ED,Sotiroudis TG,Papadimitriou V,Xenakis A,2009.Partial purification and characterization of peroxidase from olives(Olea europaea cv.Koroneiki).European Food Research and Technology,228:487-495.
Yu GC,Wen XH,Li R,Qian Y,2006.In vitro degradation of a reactive azo dye by crude ligninolytic enzymes from nonimmersed liquid culture of Phanerochaete chrysosporium.Process Biochemistry,41:1987-1993.