Youn Jung-Won, Jolkver Elena, Krämer Reinhard, Marin Kay, Wendisch Volker F
Institute of Molecular Microbiology and Biotechnology, Westfalian Wilhelms University Muenster, Corrensstr. 3, D-48149 Muenster, Germany.
J Bacteriol. 2008 Oct;190(19):6458-66. doi: 10.1128/JB.00780-08. Epub 2008 Jul 25.
Many bacteria can utilize C(4)-carboxylates as carbon and energy sources. However, Corynebacterium glutamicum ATCC 13032 is not able to use tricarboxylic acid cycle intermediates such as succinate, fumarate, and l-malate as sole carbon sources. Upon prolonged incubation, spontaneous mutants which had gained the ability to grow on succinate, fumarate, and l-malate could be isolated. DNA microarray analysis showed higher mRNA levels of cg0277, which subsequently was named dccT, in the mutants than in the wild type, and transcriptional fusion analysis revealed that a point mutation in the promoter region of dccT was responsible for increased expression. The overexpression of dccT was sufficient to enable the C. glutamicum wild type to grow on succinate, fumarate, and l-malate as the sole carbon sources. Biochemical analyses revealed that DccT, which is a member of the divalent anion/Na(+) symporter family, catalyzes the effective uptake of dicarboxylates like succinate, fumarate, L-malate, and likely also oxaloacetate in a sodium-dependent manner.
许多细菌能够利用C(4)-羧酸盐作为碳源和能源。然而,谷氨酸棒杆菌ATCC 13032不能将三羧酸循环中间体(如琥珀酸、富马酸和L-苹果酸)用作唯一碳源。经过长时间培养,可以分离出获得在琥珀酸、富马酸和L-苹果酸上生长能力的自发突变体。DNA微阵列分析表明,突变体中cg0277(随后命名为dccT)的mRNA水平高于野生型,转录融合分析表明dccT启动子区域的一个点突变导致表达增加。dccT的过表达足以使谷氨酸棒杆菌野生型能够以琥珀酸、富马酸和L-苹果酸作为唯一碳源生长。生化分析表明,DccT是二价阴离子/Na(+)同向转运体家族的成员,它以钠依赖的方式催化琥珀酸、富马酸、L-苹果酸以及可能还有草酰乙酸等二羧酸的有效摄取。