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大肠杆菌arcA突变体:以甘油作为碳源的微需氧培养物的代谢谱特征分析

Escherichia coli arcA mutants: metabolic profile characterization of microaerobic cultures using glycerol as a carbon source.

作者信息

Nikel Pablo I, Pettinari M Julia, Ramírez M Cecilia, Galvagno Miguel A, Méndez Beatriz S

机构信息

Departamento de Química Biológica, Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires, Buenos Aires, Argentina.

出版信息

J Mol Microbiol Biotechnol. 2008;15(1):48-54. doi: 10.1159/000111992. Epub 2008 Mar 14.

Abstract

ArcA is a global regulator that switches on the expression of fermentation genes and represses the aerobic pathways when Escherichia coli enters low oxygen growth conditions. The metabolic profile of E. coli CT1062 (DeltaarcA)and CT1061 (arcA2) grown in microaerobiosis with glycerol as carbon source were determined and compared with E. coli K1060, the arcA+ parent strain. Both arcA mutants achieved higher biomass yields than the wild-type strain. The production of acetate, formate, lactate, pyruvate, succinate and ethanol were determined in the supernatants of cultures grown on glycerol under microaerobic conditions for 48 h. The yield of extracellular metabolites on glycerol showed lower acid and higher ethanol values for the mutants. The ethanol/acetate ratio was 0.87 for the parent strain, 2.01 for CT1062, and 12.51 for CT1061. Accordingly, the NADH/NAD+ ratios were 0.18, 0.63, and 0.97, respectively. The extracellular succinate yield followed a different pattern, with yield values of 0.164 for K1060, 0.442 for CT1062 and 0.214 for CT1061. The dissimilarities observed can be attributed to the different effects exerted by the deletion and point mutations in a global regulator.

摘要

ArcA是一种全局调节因子,当大肠杆菌进入低氧生长条件时,它会开启发酵基因的表达并抑制需氧途径。测定了以甘油为碳源在微需氧条件下生长的大肠杆菌CT1062(ΔarcA)和CT1061(arcA2)的代谢谱,并与arcA+亲本菌株大肠杆菌K1060进行了比较。两个arcA突变体的生物量产量均高于野生型菌株。在微需氧条件下以甘油为培养基培养48小时后,测定培养上清液中乙酸、甲酸、乳酸、丙酮酸、琥珀酸和乙醇的产量。突变体在甘油上的细胞外代谢产物产量显示出较低的酸值和较高的乙醇值。亲本菌株的乙醇/乙酸比率为0.87,CT1062为2.01,CT1061为12.51。相应地,NADH/NAD+比率分别为0.18、0.63和0.97。细胞外琥珀酸产量呈现出不同的模式,K1060的产量值为0.164,CT1062为0.442,CT1061为0.214。观察到的差异可归因于全局调节因子中的缺失和点突变所产生的不同影响。

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