作　　者： ; ; ; ;

机构地区： 南宁邦尔克生物技术有限责任公司广西南宁530003

出　　处： 《广西大学学报（自然科学版）》 2008年第3期317-321,332,共6页

摘　　要： 建立一种快捷的检测基因工程菌代谢丙酮酸能力的方法,对于开展与丙酮酸相关的代谢流研究至关重要.本研究基于丙酮酸与2,4-二硝基苯肼发生羰基化反应的原理,建立了优化的丙酮酸检测体系;研究了葡萄糖等18种代谢相关物质对所建检测体系的影响;成功地将之推广到对发酵产物体系的检测中;利用含有丙酮酸甲酸裂解酶基因的大肠杆菌BL21工程菌发酵以丙酮酸为唯一碳源的培养基,结果进一步验证了该检测方法切实可行. A simple and useful method for studying engineered strain metabolizing pyruvic acid is critical to metabolism flux research in the metabolism engineering and modern gene engineering. Based on the carbonylation between pyruvic acid and 2, 4- Dinitrophenylhydrazine, after clarifying the influcence of pH,temperature and reagent ratio, a method considering the optimal constitutes and reaction conditions was constructed. The common 18 materials found in culture medium, TCA cycle or fermentation products hardly interfere the carbonylation between pyruvic acid and 2,4-Dinitrophenylhydrazine. The ability to consume pyruvid acid of engineered E. coli BL21 （DE3）pLysS containing pET22b-pflAB was successfule studied further proved the new method.

关 键 词： 丙酮酸 二硝基苯肼 基因工程菌 丙酮酸甲酸裂解酶 羰基化反应

领　　域： [生物学]