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Genetic Characterization of the Resorcinol Catabolic Pathway in Corynebacterium glutamicum

机译:谷氨酸棒杆菌中间苯二酚分解代谢途径的遗传特征

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Corynebacterium glutamicum grew on resorcinol as a sole source of carbon and energy. By genome-wide data mining, two gene clusters, designated NCgl1110-NCgl1113 and NCgl2950-NCgl2953, were proposed to encode putative proteins involved in resorcinol catabolism. Deletion of the NCgl2950-NCgl2953 gene cluster did not result in any observable phenotype changes. Disruption and complementation of each gene at NCgl1110-NCgl1113, NCgl2951, and NCgl2952 indicated that these genes were involved in resorcinol degradation. Expression of NCgl1112, NCgl1113, and NCgl2951 in Escherichia coli revealed that NCgl1113 and NCgl2951 both coded for hydroxyquinol 1,2-dioxygenases and NCgl1112 coded for maleylacetate reductases. NCgl1111 encoded a putative monooxygenase, but this putative hydroxylase was very different from previously functionally identified hydroxylases. Cloning and expression of NCgl1111 in E. coli revealed that NCgl1111 encoded a resorcinol hydroxylase that needs NADPH as a cofactor. E. coli cells containing Ncgl1111 and Ncgl1113 sequentially converted resorcinol into maleylacetate. NCgl1110 and NCgl2950 both encoded putative TetR family repressors, but only NCgl1110 was transcribed and functional. NCgl2953 encoded a putative transporter, but disruption of this gene did not affect resorcinol degradation by C. glutamicum. The function of NCgl2953 remains unclear.
机译:谷氨酸棒杆菌在间苯二酚上生长,是碳和能量的唯一来源。通过全基因组数据挖掘,提出了两个基因簇,分别命名为NCg1110-NCgl1313和NCgl2950-NCgl2953,以编码与间苯二酚分解代谢有关的推定蛋白质。 NCgl2950-NCgl2953基因簇的删除不会导致任何可观察到的表型变化。 NCg1110-NCgl1113,NCgl2951和NCgl2952上每个基因的破坏和互补表明这些基因与间苯二酚降解有关。 NCgl1112,NCgl1113和NCgl2951在大肠杆菌中的表达表明,NCgl1113和NCgl2951均编码羟基喹啉1,2-二加氧酶,而NCgl1112均编码马来酸乙酸酯还原酶。 NCg1111编码一个推定的单加氧酶,但该推定的羟化酶与以前在功能上鉴定的羟化酶有很大不同。 NCgl1111在大肠杆菌中的克隆和表达表明,NCgl1111编码间苯二酚羟化酶,需要NADPH作为辅因子。包含Ncg1111和Ncg1113的大肠杆菌细胞将间苯二酚依次转化为马来酸乙酸酯。 NCgl1110和NCgl2950都编码推定的TetR家族阻遏物,但只有NCgl1110被转录且具有功能。 NCgl2953编码推定的转运蛋白,但该基因的破坏并不影响谷氨酸棒杆菌对间苯二酚的降解。 NCgl2953的功能仍不清楚。

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