Institute of Microbiology and Biotechnology, University of Ulm, Ulm, Germany.
J Bacteriol. 2012 Mar;194(5):941-55. doi: 10.1128/JB.06064-11. Epub 2011 Dec 16.
Expression profiling of Corynebacterium glutamicum in comparison to a derivative deficient in the transcriptional regulator AtlR (previously known as SucR or MtlR) revealed eight genes showing more than 4-fold higher mRNA levels in the mutant. Four of these genes are located in the direct vicinity of the atlR gene, i.e., xylB, rbtT, mtlD, and sixA, annotated as encoding xylulokinase, the ribitol transporter, mannitol 2-dehydrogenase, and phosphohistidine phosphatase, respectively. Transcriptional analysis indicated that atlR and the four genes are organized as atlR-xylB and rbtT-mtlD-sixA operons. Growth experiments with C. glutamicum and C. glutamicum ΔatlR, ΔxylB, ΔrbtT, ΔmtlD, and ΔsixA derivatives with sugar alcohols revealed that (i) wild-type C. glutamicum grows on D-arabitol but not on other sugar alcohols, (ii) growth in the presence of D-arabitol allows subsequent growth on D-mannitol, (iii) D-arabitol is cometabolized with glucose and preferentially utilized over D-mannitol, (iv) RbtT and XylB are involved in D-arabitol but not in D-mannitol metabolism, (v) MtlD is required for D-arabitol and D-mannitol metabolism, and (vi) SixA is not required for growth on any of the substrates tested. Furthermore, we show that MtlD confers D-arabitol and D-mannitol dehydrogenase activities, that the levels of these and also xylulokinase activities are generally high in the C. glutamicum ΔatlR mutant, whereas in the parental strain, they were high when cells were grown in the presence of D-arabitol and very low when cells were grown in its absence. Our results show that the XylB, RbtT, and MtlD proteins allow the growth of C. glutamicum on D-arabitol and that D-arabitol metabolism is subject to arabitol-dependent derepression by AtlR.
与缺失转录调节因子 AtlR(以前称为 SucR 或 MtlR)的衍生物相比,对谷氨酸棒杆菌的表达谱进行分析后,发现 8 个基因在突变体中的 mRNA 水平高出 4 倍以上。其中 4 个基因位于 atlR 基因的直接附近,即编码木酮糖激酶的 xylB、核糖醇转运蛋白 rbtT、甘露醇 2-脱氢酶 mtlD 和磷酸组氨酸磷酸酶 sixA。转录分析表明,atlR 和这 4 个基因组成 atlR-xylB 和 rbtT-mtlD-sixA 操纵子。用糖醇进行谷氨酸棒杆菌和谷氨酸棒杆菌ΔatlR、ΔxylB、ΔrbtT、ΔmtlD 和ΔsixA 衍生物的生长实验表明:(i)野生型谷氨酸棒杆菌可以生长在 D-阿拉伯糖醇上,但不能生长在其他糖醇上;(ii)在 D-阿拉伯糖醇存在的情况下生长,可以随后在 D-甘露醇上生长;(iii)D-阿拉伯糖醇与葡萄糖共代谢,优先于 D-甘露醇被利用;(iv)RbtT 和 XylB 参与 D-阿拉伯糖醇但不参与 D-甘露醇代谢;(v)MtlD 是 D-阿拉伯糖醇和 D-甘露醇代谢所必需的;(vi)SixA 对于测试的任何底物的生长都不是必需的。此外,我们还表明,MtlD 赋予 D-阿拉伯糖醇和 D-甘露醇脱氢酶活性,这些酶的水平以及木酮糖激酶活性在谷氨酸棒杆菌ΔatlR 突变体中通常较高,而在亲株中,当细胞在 D-阿拉伯糖醇存在下生长时,这些酶的水平较高,而当细胞在 D-阿拉伯糖醇不存在下生长时,这些酶的水平非常低。我们的结果表明,XylB、RbtT 和 MtlD 蛋白允许谷氨酸棒杆菌在 D-阿拉伯糖醇上生长,并且 D-阿拉伯糖醇代谢受 AtlR 调控的阿拉伯糖醇依赖性去阻遏控制。