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球形红细菌生物转化2,4-二硝基甲苯及硝基还原酶性质
Biotransformation of 2,4-Dinitrotoluene and Properties of Nitroreductase by Rhodobacter Sphaeroides
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中文摘要:
      为了探索球形红细菌生物转化2,4-二硝基甲苯(2,4-DNT)的中间产物变化趋势及硝基还原酶性质,研究了不同氮源组合对球形红细菌生物转化2,4-DNT的效率及球形红细菌细胞生长的影响,测定了2,4-DNT及2,4-DNT生物转化产物在不同时间的浓度变化, 用非变性聚丙烯酰胺凝胶电泳(native-PAGE)法分析了硝基还原酶的同工酶谱,测得了硝基还原酶的动力学参数。结果表明,球形红细菌转化2,4-DNT的最适组合氮源为(NH4)2SO4/酵母膏; 球形红细菌能够高效还原2,4-DNT为4-氨基-2-硝基甲苯(4A2NT)和2-氨基-4-硝基甲苯(2A4NT),但4A2NT的含量远大于2A4NT的含量,它们没有进一步还原为2,4-二氨基甲苯(2,4-DAT)。当2,4-DNT初始浓度为20、40和60 mg·L-1时,对球形红细菌产生硝基还原酶活力有明显的促进作用,当2,4-DNT初始浓度为80 mg·L-1时,对该酶活力有抑制作用; 当pH值为7时,球形红细菌产生的硝基还原酶活力最高。球形红细菌中硝基还原酶的动力学参数,最大反应速率vmax和米氏常数Km,分别为0.0507 mmol/(L·min)和0.1772 mmol·L-1
中文关键词: 球形红细菌  2,4-二硝基甲苯(2,4-DNT)  氮源  生物转化产物  硝基还原酶  同工酶  聚丙烯酰胺凝胶电泳
英文摘要:
      To explore the change trend of intermediate products in biotransformation of 2,4-dinitrotoluene (2,4-DNT) and properties of nitroreductase produced by Rhodobacter sphaeroides, the effect of different combination of nitrogen sources on the efficiency of biotransformation 2,4-DNT by Rhodobacter sphaeroides and the growth of Rhodobacter sphaeroides cells was studied. The concentration changes of 2,4-DNT and biotransformation products of 2,4-DNT were measured at different time. The patterns of isoenzymes of nitroreductase was analyzed using native-polyacrylamide gel elecrophoresis (native-PAGE) method, and kinetic parameters of the nitroreductase were measured and obtained. Results show that the optimum combination of nitrogen sources to biotransformation of 2,4-DNT by Rhodobacter sphaeroides is complex nitrogen sources composed of yeast extract and (NH4)2SO4. Using Rhodobacter sphaeroides, 2,4-DNT can efficiently reduced as 4-amino-2-nitrotoluene(4A2NT)and 2-amino-4-nitrotoluene(2A4NT). However, the content of 4A2NT is far greater than that of 2A4NT, they have not been futher reduced to 2,4-diaminotoluene (2,4-DAT). When the initial concentration of 2,4-DNT is 20,40 and 60 mg·L-1, there is an obvious promoting effect on the actiyity of nitroreductas produced by Rhodobacter sphaeroides. While, when the initial concentration of 2,4-DNT is 80 mg·L-1, the actiyity of nitroreductase is inhibited. The activity of nitroreductas produced by Rhodobacter sphaeroides is the highest at the initial pH=7. The kinetic parameters, the maximum reaction rate Vmax and Michaelis constant Km, for the nitroreductase in Rhodobacter sphaeroides are 0.0507 mmol·(L·min) -1 and 0.1772 mmol·L-1, respectively.
英文关键词: Rhodobacter sphaeroides  2,4-dinitrotoluene  nitrogen source  biotransformation products  nitroreductase  isoenzyme  polyacrylamide gel elecrophoresis
DOI:10.11943/j.issn.1006-9941.2018.03.013
投稿日期:2017-09-19
最后修改日期:2017-11-15
网络出版日期:2018-03-13
书版出版日期:2018-03-21
基金项目:山西省回国留学人员科研资助项目(2016-084)
引用文献:王寿艳,白红娟,赵婷婷,康鹏洲,等.球形红细菌生物转化2,4-二硝基甲苯及硝基还原酶性质[J].含能材料,2018,26(3):279~284
作者单位E-mail
王寿艳 中北大学环境与安全工程学院, 山西 太原 030051 1345406573@qq.com,bhj44871@163.com 
白红娟 中北大学环境与安全工程学院, 山西 太原 030051 bhj44871@163.com 
赵婷婷 中北大学环境与安全工程学院, 山西 太原 030051  
康鹏洲 中北大学环境与安全工程学院, 山西 太原 030051  
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