Kurthia huakuii LAM0618~T重组酚氧化酶LaclK对乙基紫脱色的研究
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摘要
以三苯甲烷类染料乙基紫为研究对象,研究了Kurthia huakuii LAM0618~T重组酚氧化酶LaclK对乙基紫的脱色性能,探讨了给酶量、脱色时间、染料浓度、反应p H、卤离子、有机试剂以及NO~-_3、SO~(2-)_4和腐殖酸对脱色效果的影响。在pH 7.0,60℃条件下反应1 h,200 U/L的LaclK对10μmol/L和20μmol/L的乙基紫显示出较好的脱色效果,脱色率分别达到91%和84%。此外,LaclK能够耐受卤离子和有机试剂,具有一定的生物修复应用潜力。
The dye decolorization ability of recombinant Lacl K on ethyl violet was studied in this work. Effects of parameters,such as amount of Lacl K,incubation time,ethyl violet concentration,p H,halide ions,organic solvents,NO~-_3,SO~(2-)_4 and humic acid on the decolorization was investigated. After incubation at 60℃ and pH 7. 0 for 1 h,200 U / L of Lacl K decolorized 91% of 10 μmol / L ethyl violet and 84% of 20 μmol / L ethyl violet,respectively. Moreover,Lacl K exhibited a strong tolerance to halide ions and organic solvents,indicating the industrial application potential on bioremediation.
The dye decolorization ability of recombinant Lacl K on ethyl violet was studied in this work. Effects of parameters,such as amount of Lacl K,incubation time,ethyl violet concentration,p H,halide ions,organic solvents,NO~-_3,SO~(2-)_4 and humic acid on the decolorization was investigated. After incubation at 60℃ and pH 7. 0 for 1 h,200 U / L of Lacl K decolorized 91% of 10 μmol / L ethyl violet and 84% of 20 μmol / L ethyl violet,respectively. Moreover,Lacl K exhibited a strong tolerance to halide ions and organic solvents,indicating the industrial application potential on bioremediation.
引文
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[14]Fang Z,Li T,Wang Q,et al..A bacterial laccase from marine microbial metagenome exhibiting chloride tolerance and dye decolorization ability[J].Appl.Microbiol.Biotechnol.,2011,89:1103-1110.
[15]Saratale R G,Saratale G D,Chang J S,et al..Bacterial decolorization and degradation of azo dyes:A review[J].J.Taiwan.Inst.Chem.Eng.,2011,42(1):138-157.
[16]Baldrian P.Fungal laccases-occurrence and properties[J].FEMS.Microbiol.Rev.,2006,30:215-242.
[17]Liu H,Cheng Y,Du B,et al..Overexpression of a novel thermostable and chloride-tolerant laccase from Thermus thermophilus SG0.5JP17-16 in Pichia pastoris and its application in synthetic dye decolorization[J].PLo S ONE,2015,10(3):e0119833.
[18]Singh G,Sharma P,Capalash N.Performance of an alkalophilic and halotolerant laccase fromγ-proteobacterium JB in the presence of industrial pollutants[J].J.Gen.Appl.Microbiol.,2009,55:283-289.
[19]Khmelnitsky Y L,Rich J O.Biocatalysis in nonaqueous solvents[J].Curr.Opin.Chem.Biol.,1994,3:47-53.
[20]Klibanov A M.Why are enzymesless active in organic solvents than in water?[J].Trends Biotechnol.,1997,15:97-101.
[21]Erdemoglu S,Aksu S K,Sayilkan F,et al..Photocatalytic degradation of Congo Red by hydrothermally synthesized nanocrystalline Ti O2and identification of degradation products by LC-MS[J].J.Hazard.Mater.,2008,155(3):469-476.
[22]Kumar P,Prasad B,Mishra I,et al..Catalytic thermal treatment of desizing wastewaters[J].J.Hazard.Mater.,2007,149:26-34.
[23]Sinirlioglu Z A,Sinirlioglu D,Akbas F.Preparation and characterization of stable cross-linked enzyme aggregates of novel laccase enzyme from Shewanella putrefaciens and using malachite green decolorization[J].Bioresour.Technol.,2013,146:807-811.
[24]Rasekh B,Khajeh K,Ranjbar B,et al..Protein engineering of laccase to enhance its activity and stability in the presence of organic solvents[J].Engin.Life Sci.,2014,14(4):442-448.
[2]Selvam K,Swaminathan K,Chae K S.Decolourization of azo dyes and a dye industry effluent by a white rot fungus Thelephora sp.[J].Bioresour.Technol.,2003,88(2):115-119.
[3]TunS,Gürkan T,Duman O.On-line spectrophotometric method for the determination of optimum operation parameters on the decolorization of acid red 66 and direct blue 71 from aqueous solution by fenton process[J].Chem.Eng.J.,2012,(181-182):431-442.
[4]Shah M P.Microbe-mediated degradation of synthetic dyes in wastewater[A].In:Singh S N.Microbial Degradation of Synthetic Dyes in Wastewaters[M].Springer International Publishing Switzerland,2015,205-242.
[5]Gao F,Ding H,Shao L,et al..Molecular characterization of a novel thermal stable reductase capable of decoloration of both azo and triphenylmethane dyes[J].Appl.Microbiol.Biotechnol.,2015,99:255-267.
[6]Jasin ska A,Paraszkiewicz K,Saba M,et al..Microbial decolorization of triphenylmethane dyes[A].In:Singh S N.Microbial Degradation of Synthetic Dyes in Wastewaters[M].Springer International Publishing Switzerland,2015,169-186.
[7]Singh L,Singh V P.Textile dyes degradation:a microbial approach for biodegradation of pollutants[A].In:Singh S N.Microbial Degradation of Synthetic Dyes in Wastewaters[M].Springer International Publishing Switzerland,2015,187-204.
[8]张培培,任随周,许玫英,等.微生物对三苯基甲烷类染料脱色的研究进展[J].微生物学通报,2009,32(9):1410-1417.
[9]Jasinska A,Paraszkiewicz K,Sip A,et al..Malachite green decolorization by the filamentous fungus Myrothecium roridummechanistic study and process optimization[J].Bioresour.Technol.,2015,194:43-48.
[10]Sani R K,Banerjee U C.Decolorization of triphenylmethane dyes and textile and dye-stuff effluent by Kurthia sp.[J].Enzyme Microb.Technol.,1999,24(7):433-437.
[11]Ruan Z,Wang Y,Song J,et al..Kurthia huakuii sp.nov.,isolated from biogas slurry,and emended description of the genus Kurthia[J].Int.J.Syst.Evol.Microbiol.,2014,64:518-521.
[12]Cong W,Zhu Z,He H,et al..A visible dye-based staining method for DNA in polyacrylamide gels by ethyl violet[J].Anal.Biochem.,2010,402(1):99-101.
[13]Velusamy P,Pitchaimuthu S,Rajalakshmi S,et al..Modification of the photocatalytic activity of Ti O2by betacyclodextrin in decoloration of ethyl violet dye[J].J.Adv.Res.,2014,5(1):19-25.
[14]Fang Z,Li T,Wang Q,et al..A bacterial laccase from marine microbial metagenome exhibiting chloride tolerance and dye decolorization ability[J].Appl.Microbiol.Biotechnol.,2011,89:1103-1110.
[15]Saratale R G,Saratale G D,Chang J S,et al..Bacterial decolorization and degradation of azo dyes:A review[J].J.Taiwan.Inst.Chem.Eng.,2011,42(1):138-157.
[16]Baldrian P.Fungal laccases-occurrence and properties[J].FEMS.Microbiol.Rev.,2006,30:215-242.
[17]Liu H,Cheng Y,Du B,et al..Overexpression of a novel thermostable and chloride-tolerant laccase from Thermus thermophilus SG0.5JP17-16 in Pichia pastoris and its application in synthetic dye decolorization[J].PLo S ONE,2015,10(3):e0119833.
[18]Singh G,Sharma P,Capalash N.Performance of an alkalophilic and halotolerant laccase fromγ-proteobacterium JB in the presence of industrial pollutants[J].J.Gen.Appl.Microbiol.,2009,55:283-289.
[19]Khmelnitsky Y L,Rich J O.Biocatalysis in nonaqueous solvents[J].Curr.Opin.Chem.Biol.,1994,3:47-53.
[20]Klibanov A M.Why are enzymesless active in organic solvents than in water?[J].Trends Biotechnol.,1997,15:97-101.
[21]Erdemoglu S,Aksu S K,Sayilkan F,et al..Photocatalytic degradation of Congo Red by hydrothermally synthesized nanocrystalline Ti O2and identification of degradation products by LC-MS[J].J.Hazard.Mater.,2008,155(3):469-476.
[22]Kumar P,Prasad B,Mishra I,et al..Catalytic thermal treatment of desizing wastewaters[J].J.Hazard.Mater.,2007,149:26-34.
[23]Sinirlioglu Z A,Sinirlioglu D,Akbas F.Preparation and characterization of stable cross-linked enzyme aggregates of novel laccase enzyme from Shewanella putrefaciens and using malachite green decolorization[J].Bioresour.Technol.,2013,146:807-811.
[24]Rasekh B,Khajeh K,Ranjbar B,et al..Protein engineering of laccase to enhance its activity and stability in the presence of organic solvents[J].Engin.Life Sci.,2014,14(4):442-448.