苯并(a)芘降解过程中细菌降解相关酶的蛋白应答
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摘要
以苯并(a)芘(BaP)为目标污染物,探讨了3株细菌(BZSY-1、BZSY-2为芽孢杆菌; BZSY-3为动胶杆菌)在不同条件下对BaP细菌降解的酶蛋白应答以及降解率与酶活性的关系。结果表明:不同浓度BaP降解过程中,3株细菌的邻苯二酚2,3-双加氧酶(C23O)酶活性均较高,多酚氧化酶(PPO)酶活性均较低,而过氧化氢酶(CAT)酶活性与BaP的浓度无关;加入共代谢底物琥珀酸钠后,3株细菌的邻苯二酚1,2-双加氧酶(C12O)、C23O酶活性明显提高,CAT酶活性无明显变化,而PPO酶活性通过诱导依然很低;与对照相比,3株菌的C12O、C23O和PPO在TW80≤1000μg·m L~(-1)时酶活性不受影响,而当TW80>2000μg·m L~(-1)时,3株细菌的这3种酶均受到不同程度的抑制;与细菌BZSY-1相比,BZSY-2和BZSY-3对BaP的降解率较高,对应的几种酶活性(尤其是C12O、C23O)也比较高,进一步支持BaP的微生物降解是酶促反应,这3株细菌对BaP降解的关键酶是C12O和C23O。
The protein response of bacterial enzymes involved in degradation of benzo( a) pyrene( BaP) by three bacteria( BZSY-1,BZSY-2 for Bacillus,BZSY-3 for Zooloea sp.) were examined under different conditions. The relationships between the degradation rate and enzyme activity were analyzed. Four enzymes( C12O,C23O,CAT and PPO) were detected upon using different concentrations of BaP. Among the four enzymes,the activity of C23O was the highest and that of PPO was the lowest. The activity of CAT was not correlated with the concentration of BaP.After adding sodium succinate( co-metabolites),the activities of C12O and C23O were enhanced,whereas that of CAT was not changed,and the activity of PPO was still lower. Compared with the control,the activities of C12O,C23O and PPO were not affected when the concentration of TW80 was less than or equal to 1000 μg·mL~(-1). When the concentration of TW80 was greater than 2000 μg · m L~(-1),the activities of these three enzymes were inhibited by various degrees.Compared with BZSY-1,both BZSY-2 and BZSY-3 had higher BaP degradation rates and higher activities of the corresponding enzymes( especially C12O and C23O),which further supported the notion that the microbial degradation of BaP is an enzymatic reaction. C12O and C23O are the two key BaP-degrading enzymes in the three bacteria.
The protein response of bacterial enzymes involved in degradation of benzo( a) pyrene( BaP) by three bacteria( BZSY-1,BZSY-2 for Bacillus,BZSY-3 for Zooloea sp.) were examined under different conditions. The relationships between the degradation rate and enzyme activity were analyzed. Four enzymes( C12O,C23O,CAT and PPO) were detected upon using different concentrations of BaP. Among the four enzymes,the activity of C23O was the highest and that of PPO was the lowest. The activity of CAT was not correlated with the concentration of BaP.After adding sodium succinate( co-metabolites),the activities of C12O and C23O were enhanced,whereas that of CAT was not changed,and the activity of PPO was still lower. Compared with the control,the activities of C12O,C23O and PPO were not affected when the concentration of TW80 was less than or equal to 1000 μg·mL~(-1). When the concentration of TW80 was greater than 2000 μg · m L~(-1),the activities of these three enzymes were inhibited by various degrees.Compared with BZSY-1,both BZSY-2 and BZSY-3 had higher BaP degradation rates and higher activities of the corresponding enzymes( especially C12O and C23O),which further supported the notion that the microbial degradation of BaP is an enzymatic reaction. C12O and C23O are the two key BaP-degrading enzymes in the three bacteria.
引文
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钟鸣,周启星,许华夏,等.2004.降解性细菌对菲诱导的蛋白及酶活性应答反应.应用生态学报,15(5):871-874.
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Suzuki N,Sato M,Nassar HF,et al.2016.Seawater polluted with highly concentrated polycyclic aromatic hydrocarbons suppresses osteoblastic activity in the scales of goldfish,Carassius auratus.Zoological Science,33:407-413.
Wei R,Ni J,Li X,et al.2017.Dissipation and phytoremediation of polycyclic aromatic hydrocarbons in freshly spiked and long-term field-contaminated soils.Environmental Science&Pollution Research,24:7994-8003.
Zang S,Lian B.2009.Synergistic degradation of 2-naphthol by Fusarium proliferatum and Bacillus subtilis in wastewater.Journal of Hazardous Materials,166:33-38.
李忠光,龚明.2005.植物多酚氧化酶活性测定方法的改进.云南师范大学学报:自然科学版,25(1):44-45.
刘洋.2017.双加氧酶催化邻苯二酚类化合物生物降解机理的研究(博士学位论文).广州:华南理工大学.
刘建国,唐孝炎,胡建信.2003.持久性生物累积性有毒污染物与国际相关控制策略和行动.环境保护,(4):52-56.
曲春香,沈颂东,王雪峰,等.2006.用考马斯亮蓝测定植物粗提液中可溶性蛋白质含量方法的研究.苏州大学学报:自然科学版,22(2):82-85.
宋宇,王健,苏振成,等.2016.PAHs污染土壤的物理强化修复过程中土壤细菌的响应.生态学杂志,35(6):1547-1552.
张小凡,柳津广志.2003.多环芳烃化合物菲分解菌的分离鉴定及分解特性研究.上海环境科学,(8):544-547.
张银萍,王芳,杨兴伦,等.2010.土壤中高环多环芳烃微生物降解的研究进展.微生物学通报,37(2):280-288.
钟鸣,周启星,许华夏,等.2004.降解性细菌对菲诱导的蛋白及酶活性应答反应.应用生态学报,15(5):871-874.
朱婷婷,倪晋仁.2012.不同碳源对苯并[a]芘降解菌生长和降解性能影响的研究.北京大学学报:自然科学版,48(2):343-346.
And SHC,Aitken MD.1999.Salicylate stimulates the degradation of high-molecular weight polycyclic aromatic hydrocarbons by Pseudomonas saccharophila P15.Environmental Science&Technology,33:435-439.
Cerniglia CE,Gibson DT.1979.Oxidation of benzo[a]pyrene by the filamentous fungus Cunninghamella elegans.Journal of Biological Chemistry,254:12174-12180.
Ijoma GN,Tekere M.2017.Potential microbial applications of co-cultures involving ligninolytic fungi in the bioremediation of recalcitrant xenobiotic compounds.International Journal of Environmental Science and Technology,14:1-20.
Keith LH,Telliard WA.1979.Priority pollutants:I.A perspective view.Environmental Science&Technology,13:416-423.
Pongpiachan S,Hattayanone M,Pinyakong O,et al.2016.Quantitative ecological risk assessment of inhabitants exposed to polycyclic aromatic hydrocarbons in terrestrial soils of King George Island,Antarctica.Polar Science,11:19-29.
Romero MC,Salvioli ML,Cazau MC,et al.2002.Pyrene degradation by yeasts and filamentous fungi.Environmental Pollution,117:159-163.
Suzuki N,Sato M,Nassar HF,et al.2016.Seawater polluted with highly concentrated polycyclic aromatic hydrocarbons suppresses osteoblastic activity in the scales of goldfish,Carassius auratus.Zoological Science,33:407-413.
Wei R,Ni J,Li X,et al.2017.Dissipation and phytoremediation of polycyclic aromatic hydrocarbons in freshly spiked and long-term field-contaminated soils.Environmental Science&Pollution Research,24:7994-8003.
Zang S,Lian B.2009.Synergistic degradation of 2-naphthol by Fusarium proliferatum and Bacillus subtilis in wastewater.Journal of Hazardous Materials,166:33-38.