高活性漆酶菌株6BS筛选及其对染料脱色的影响
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摘要
为了寻找能有效降解染料的菌株,以Cu2+为筛选剂,丁香醛连氮为底物从土壤样品中筛选具有较高漆酶活性的菌株,结合形态学、生理生化特性以及16SrDNA序列同源性分析等方法进行鉴定。结果表明:经鉴定土壤菌株为芽孢杆菌属细菌,命名为Bacillus sp.6BS。菌株6BS的最适生长温度和pH分别为37℃和8.0,能够耐受5%的NaCl和2mmol/L的Cu2+。菌株6BS的芽孢漆酶在不含介体的情况下对结晶紫脱色效果较好,6h脱色率可达到92%。介体乙酰丁香酮(AS)和2,2,6,6,-四甲基哌啶氧化物(TEM-PO)能显著提高菌株6BS的芽孢漆酶对靛红、活性黑和RBBR的脱色率,丁香醛(SA)能显著提高芽孢漆酶对靛红和活性黑的脱色率。天然介体AS具有非常显著的助推作用,是可推荐的介体。6BS菌株漆酶在介体的作用下具有较强的染料处理能力,是值得推荐的高效菌株。
The strain with a high laccase activity was screened from the soil sample containing syringaldazine and identified by using morphology,physiological-biochemical characteristics and 16 SrDNA sequence homology analysis to search strains with a capacity of effective dye decolorization.Result:The screened strain is identified as Bacillus 6 BS.The optimum growth temperature and pH of strain 6 BS are37℃ and 8.0 respectively.The strain 6 BS can tolerate 5% NaCl and 2 mmol/L Cu2+.The spore laccase of strain 6 BS has a good decolorization effect to crystal violet without any mediators and the decolorization rate can reach 92%after 6 h.The spore laccase of strain 6 BS can significantly increase the decolorization rate of isatin,reactive black and RBBR when acetosyringone(AS)and 2,2,6,6,-tetramethylpiperidine oxide(TEMPO)are used as the mediators.The spore laccase of strain 6 BS can improve the decolorization rate of isatin and reactive black when SA is used as the mediator significantly.The natural mediator AS with a very significant role is a recommendable mediator.The spore laccase of strain 6 BS has a strong processing capacity to dyes under the mediator role,which indicates that strain 6 BS is a recommendable high-efficiency strain.
The strain with a high laccase activity was screened from the soil sample containing syringaldazine and identified by using morphology,physiological-biochemical characteristics and 16 SrDNA sequence homology analysis to search strains with a capacity of effective dye decolorization.Result:The screened strain is identified as Bacillus 6 BS.The optimum growth temperature and pH of strain 6 BS are37℃ and 8.0 respectively.The strain 6 BS can tolerate 5% NaCl and 2 mmol/L Cu2+.The spore laccase of strain 6 BS has a good decolorization effect to crystal violet without any mediators and the decolorization rate can reach 92%after 6 h.The spore laccase of strain 6 BS can significantly increase the decolorization rate of isatin,reactive black and RBBR when acetosyringone(AS)and 2,2,6,6,-tetramethylpiperidine oxide(TEMPO)are used as the mediators.The spore laccase of strain 6 BS can improve the decolorization rate of isatin and reactive black when SA is used as the mediator significantly.The natural mediator AS with a very significant role is a recommendable mediator.The spore laccase of strain 6 BS has a strong processing capacity to dyes under the mediator role,which indicates that strain 6 BS is a recommendable high-efficiency strain.
引文
[1]MAYER A M,STAPLES R C.Laccase:new functions for an old enzyme[J].Phytochemistry,2002,60(6):551-565.
[2]GIARDINA P,FARACO V,PEZZELLA C,et al.Laccases:A never-ending story[J].Cell Mol Life Sci,2010,67:369-385.
[3]THURSTON C F.The structure and function of fungal laccases[J].Microbiology,1994,140(1):19-26.
[4]HOEGGER P J,KILARU S,JAMES T Y,et al.Phylogenetic comparison and classification of laccase and related multicopper oxidase protein sequences[J].FEBS,2006,273(10):2308-2326.
[5]司静,李伟,崔宝凯,等.真菌漆酶性质、分子生物学及其应用研究进展[J].生物技术通报,2011(2):48-55.
[6]PALMIERI G,CENNAMO G,SANNIA G.Remazol Brilliant R decolourisation by the fungi Pleurotus ostreatus and its oxidative enzymatic system[J].Enzyme Microb Technol,2005,36:17-27.
[7]施新花,邢瑞乐.印染生产工艺废水特性浅析[J].污染防治技术,2006,19(4):50-52.
[8]宋娜,王晓义.电化学技术在废水处理中的应用[J].化工管理,2012(S1):35-37.
[9]孙颖,雷彩虹.染料废水脱色的物理化学处理技术[J].中国化工管理干部学院学报,2006(4):86-88.
[10]周稳,管政兵,蔡宇杰,等.芽孢杆菌漆酶的研究进展[J].微生物学报,2015,42(7):1372-1383.
[11]王锋,刘英,汪印,等.漆酶及其应用[J].生物加工过程,2012(4):70-76.
[12]汪春蕾,李凡姝,孙海琼,等.Bacillus细菌的分离及其芽孢漆酶对染料脱色的影响[J].湖南农业大学学报(自然科学版),2016,42(1):85-90.
[13]东秀珠,蔡妙英.常见细菌系统鉴定手册[K].北京:北京科学出版社,2001:364-399.
[14]沈萍,范秀容,李广武.微生物学实验[M].3版.北京:高等教育出版社,1999:26-35.
[15]王远亮,杨瑞红,毛爱军,等.采用未培养技术对荷斯坦奶牛瘤胃细菌多样性进行初步分析[J].微生物学报,2005,45(6):915-919.
[16]GOLDMAN R C,TIPPER D J.Bacillus subtilis spore coats:complexity and purification of an unique polypeptide component[J].Biotechnology,1978,135(3):1091-1106.
[17]孙海琼,汪春蕾.耐铜菌株4BS的鉴定及其对染料脱色的效果[J].湖南农业大学学报(自然科学版),2016,42(4):403-408.
[18]李凡姝,刘海洋,戴绍军,等.高产漆酶菌株Bacillus sp.CLb的筛选及其对染料脱色效果的研究[J].安徽农业科学,2014,42(6):1614-1616,1654.
[19]汪春蕾,赵敏.细菌漆酶的研究[M].哈尔滨:哈尔滨工业大学出版社,2008.
[20]汪春蕾,田菲,张月颖,等.耐盐Bacillus sp.9BS的筛选及其对染料的脱色效果[J].湖南农业大学学报(自然科学版),2015,41(3):313-317.
[21]江莹,蔡亚军.细菌染料脱色机理研究进展[J].广州化工,2015(17):4-6.
[22]曹明乐,张海波,黄峰,等.漆酶的研究进展[J].山东林业科技,2012(3):106-115.
[23]王习文,詹怀宇.漆酶/介体系统生物漂白技术[J].西南造纸,2002(3):7-10.
[24]罗爽,谢天,刘忠川,等.漆酶/介体系统研究进展[J].应用于环境生物学报,2015,21(6):987-99.
[2]GIARDINA P,FARACO V,PEZZELLA C,et al.Laccases:A never-ending story[J].Cell Mol Life Sci,2010,67:369-385.
[3]THURSTON C F.The structure and function of fungal laccases[J].Microbiology,1994,140(1):19-26.
[4]HOEGGER P J,KILARU S,JAMES T Y,et al.Phylogenetic comparison and classification of laccase and related multicopper oxidase protein sequences[J].FEBS,2006,273(10):2308-2326.
[5]司静,李伟,崔宝凯,等.真菌漆酶性质、分子生物学及其应用研究进展[J].生物技术通报,2011(2):48-55.
[6]PALMIERI G,CENNAMO G,SANNIA G.Remazol Brilliant R decolourisation by the fungi Pleurotus ostreatus and its oxidative enzymatic system[J].Enzyme Microb Technol,2005,36:17-27.
[7]施新花,邢瑞乐.印染生产工艺废水特性浅析[J].污染防治技术,2006,19(4):50-52.
[8]宋娜,王晓义.电化学技术在废水处理中的应用[J].化工管理,2012(S1):35-37.
[9]孙颖,雷彩虹.染料废水脱色的物理化学处理技术[J].中国化工管理干部学院学报,2006(4):86-88.
[10]周稳,管政兵,蔡宇杰,等.芽孢杆菌漆酶的研究进展[J].微生物学报,2015,42(7):1372-1383.
[11]王锋,刘英,汪印,等.漆酶及其应用[J].生物加工过程,2012(4):70-76.
[12]汪春蕾,李凡姝,孙海琼,等.Bacillus细菌的分离及其芽孢漆酶对染料脱色的影响[J].湖南农业大学学报(自然科学版),2016,42(1):85-90.
[13]东秀珠,蔡妙英.常见细菌系统鉴定手册[K].北京:北京科学出版社,2001:364-399.
[14]沈萍,范秀容,李广武.微生物学实验[M].3版.北京:高等教育出版社,1999:26-35.
[15]王远亮,杨瑞红,毛爱军,等.采用未培养技术对荷斯坦奶牛瘤胃细菌多样性进行初步分析[J].微生物学报,2005,45(6):915-919.
[16]GOLDMAN R C,TIPPER D J.Bacillus subtilis spore coats:complexity and purification of an unique polypeptide component[J].Biotechnology,1978,135(3):1091-1106.
[17]孙海琼,汪春蕾.耐铜菌株4BS的鉴定及其对染料脱色的效果[J].湖南农业大学学报(自然科学版),2016,42(4):403-408.
[18]李凡姝,刘海洋,戴绍军,等.高产漆酶菌株Bacillus sp.CLb的筛选及其对染料脱色效果的研究[J].安徽农业科学,2014,42(6):1614-1616,1654.
[19]汪春蕾,赵敏.细菌漆酶的研究[M].哈尔滨:哈尔滨工业大学出版社,2008.
[20]汪春蕾,田菲,张月颖,等.耐盐Bacillus sp.9BS的筛选及其对染料的脱色效果[J].湖南农业大学学报(自然科学版),2015,41(3):313-317.
[21]江莹,蔡亚军.细菌染料脱色机理研究进展[J].广州化工,2015(17):4-6.
[22]曹明乐,张海波,黄峰,等.漆酶的研究进展[J].山东林业科技,2012(3):106-115.
[23]王习文,詹怀宇.漆酶/介体系统生物漂白技术[J].西南造纸,2002(3):7-10.
[24]罗爽,谢天,刘忠川,等.漆酶/介体系统研究进展[J].应用于环境生物学报,2015,21(6):987-99.
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