地衣芽孢杆菌转化间二硝基苯为间硝基苯羟胺、间硝基苯胺和间苯二胺的研究

从土壤中筛选得到1株转化间二硝基苯的芽孢杆菌,经生理生化实验及16SrDNA序列分析,确定其为地衣芽孢杆菌CGMCC2280。对含间二硝基苯的菌体转化液在不同时间取样,分离纯化得到化合物A、B和C,通过LC-MS和1 HNMR分析,同时与标准品对照,确定化合物A、B和C分别为间硝基苯羟胺、间硝基苯胺和间苯二胺。底物特异性表明,该菌对间二硝基苯和邻二硝基苯的底物转化率10h达到98%以上,但对对二硝基苯的底物转化率却不到10%。转化液中葡萄糖具有促进间二硝基苯还原的作用,在没有葡萄糖存在的情况下,转化反应速度较慢,且反应后期反应产物间苯二胺会进一步降解。在转化反应的不同时间加入丙酮终止反应并进行产物分析,发现该菌还原间二硝基苯的转化过程是先形成间硝基苯羟胺,再形成间硝基苯胺,最后形成间苯二胺。

Conversion of m-Dinitrobenzene into m-Nitro-Phenylene Hydroxylamine,m-Nitroaniline and m-Phenylenediamine by Bacillus Licheniformis

A Bacillus strain capable of converting m-dinitrobenzene was isolated from soil.Physiological and biochemical experiments and 16S rDNA sequences analysis confirmed that the isolated strain was a Bacillus licheniformis CGMCC2280.Sampling at different times of bacterial fermentation,three compounds of A,B and C were obtained.By LC-MS and 1HNMR analysis with the contrast of the standards,confirmed that A,B and C was m-nitro-phenylene hydroxylamine,m-nitroaniline and m-phenylenediamine,respectively.Substrate specificity study indicated that the bacterial strain converted 98% of m-dinitrobenzene and o-dinitrobenzene substrate in 10 h,but less than 10% of p-dinitrobenzene.It enhanced the reduction of m-dinitrobenzene when glucose was added into the conversion solution.In the system without adding glucose,the conversion rate was lower and the concentration of m-phenylenediamine continued to decrease.By adding acetone to terminate the reaction at different times and analyzing the products,it proved that the first product of the procedure was m-nitro-phenylene hydroxylamine,then m-nitroaniline and finally m-phenylenediamine.