腐殖酸对Mycobacterium sp. NJS-1降解芘的影响
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1.南京师范大学分析测试中心;2.南京师范大学化学与材料学院;3.中国科学院土壤环境与污染修复重点实验室

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国家重点研发计划重点专项


Effects of Humic acid on biodegradation of pyrene by Mycobacterium sp. NJS-1
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1.Nanjing Normal University Center for Analysis and Testing, School of Chemistry and Materials, Nanjing Normal University;2.Key Laboratory of Soil Environment and Pollution Remediation, Chinese Academy of Sciences;3.School of Chemistry and Materials, Nanjing Normal University;4.Nanjing Normal University Center for Analysis and Testing

Fund Project:

National Key Research and Development Program of China

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    摘要:

    为研究腐殖酸(Humic acid,HA)对微生物降解环境中多环芳烃(Polycyclic Aromatic Hydrocarbons , PAHs)的影响,以四环芘为典型PAHs及HA为代表性溶解性有机质,选取芘高效降解菌Mycobacterium sp.NJS-1于有无HA条件下降解芘,应用傅里叶变换回旋离子共振质谱(FT-ICR-MS), 激光共聚焦显微镜(CLSM), 及光谱技术分析降解过程中芘的残留特征、中间产物信息及菌落的变化特征。结果显示:未添加HA,芘降解率为35.33%±3.27%;添加HA后,其降解率明显提高,达到88.33%±3.40%,表明HA对高效菌Mycobacterium sp. NJS-1降解芘有明显的促进作用。首先,不添加HA,菌落Mycobacterium sp. NJS-1分泌的双加氧酶只进攻芘的C4,5位点,而添加HA后增加了进攻芘的C1,2位点,因此增加了芘的代谢途径。添加HA后,CLSM结果显示无死亡细胞出现而且团聚明显,表明HA能够提高细胞的存活率并促进微生物膜的形成。同时,在降解芘的过程中,产生了酪氨酸类蛋白质和可见光类富里酸,表明添加HA反而提高了降解菌活性,而且形成了三维结构的生物膜。光谱分析部分发现:蛋白酰胺,酪氨酸、色氨酸和苯丙氨酸发生红移,表明HA的添加引起了细胞表面官能团作用力的变化。可见降解菌不仅通过细胞表面官能团相互作用形成微生物膜促进降解,而且外源HA能够促进菌落之间的作用,加快降解的进行。 关键词:腐殖酸;芘;生物降解;微生物膜

    Abstract:

    【Objective】Pollution by Polycyclic Aromatic Hydrocarbons (PAHs) in the soil environment has become a serious problem. In recent years, microbial degradation of PAHs to achieve soil remediation has become an important method. However, microbial degradation is not a very efficient and ideal process for the remediation of PAHs-polluted environments. In this study, humic acid (HA) was added during the degradation process to explore its effect on the microbial degradation of PAHs. 【Method】 Using pyrene as a typical PAH and HA as representative dissolved organic matter, Mycobacterium sp. NJS-1 with high pyrene degradation efficiency was selected to degrade pyrene with or without HA. Fourier transform cyclotron resonance mass spectrometry (FT-ICR-MS), laser confocal microscopy (CLSM), and techniques such as infrared and Raman spectroscopies were used to analyze the residual characteristics of pyrene, intermediates and colony changes during degradation. 【Results】 Results show: (1) The presence of HA significantly accelerated pyrene biodegradation. Approximately 88.33%±3.40% of pyrene was biodegraded within 7 days of incubation with the addition of HA, but only 35.33%±3.27% was biodegraded without HA; (2) The FT-ICR-MS results indicated that addition of HA induced two biodegradation pathways - attacking of 1,2-positions and 4,5-positions of pyrene by dioxygenase, whereas in the absence of HA only the 4,5-position was attacked; (3) The CLSM results showed that HA enhanced the growth of Mycobacterium sp. NJS-1 with no death of bacteria cells. Simultaneously, tyrosine proteins and fulvic acid-like compounds under visible light were detected and showed that HA enhanced bacterial activity by biofilm formation; (4) The results of the spectral analysis showed that a series of bands, mainly including protein amide, tyrosine, tryptophan and phenylalanine, red-shifted, implying that the interacted forces from the functional groups of bacterial surfaces changed in the presence of HA.【Conclusion】 HA can improve the degradation efficiency by increasing the degradation pathway of pyrene and inducing the interaction of functional groups on the cell surface to form a microbial membrane. Meanwhile, the addition of exogenous HA could also promote the interactions among colonies and accelerate the degradation. Keywords: Humic acid; Pyrene; Biodegradable; Microbial film

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方文雯,李晓宁,吴仕希,王芳,顾正桂,张银萍.腐殖酸对Mycobacterium sp. NJS-1降解芘的影响[J].土壤学报,,[待发表]
fang wenwen, Li Xiaoning, wu shixi, wang fang, gu zhenggui, zhang yinping. Effects of Humic acid on biodegradation of pyrene by Mycobacterium sp. NJS-1[J]. Acta Pedologica Sinica,,[In Press]

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