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首页> 外文期刊>Applied Microbiology >Phenylacetic Acid Catabolism and Its Transcriptional Regulation in Corynebacterium glutamicum
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Phenylacetic Acid Catabolism and Its Transcriptional Regulation in Corynebacterium glutamicum

机译：谷氨酸棒杆菌中苯乙酸的分解代谢及其转录调控

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The industrially important organism Corynebacterium glutamicum has been characterized in recent years for its robust ability to assimilate aromatic compounds. In this study, C. glutamicum strain AS 1.542 was investigated for its ability to catabolize phenylacetic acid (PAA). The paa genes were identified; they are organized as a continuous paa gene cluster. The type strain of C. glutamicum , ATCC 13032, is not able to catabolize PAA, but the recombinant strain ATCC 13032/pEC-K18 mob 2:: paa gained the ability to grow on PAA. The paaR gene, encoding a TetR family transcription regulator, was studied in detail. Disruption of paaR in strain AS 1.542 resulted in transcriptional increases of all paa genes. Transcription start sites and putative promoter regions were determined. An imperfect palindromic motif (5′-ACTNACCGNNCGNNCGGTNAGT-3′; 22 bp) was identified in the upstream regions of paa genes. Electrophoretic mobility shift assays (EMSA) demonstrated specific binding of PaaR to this motif, and phenylacetyl coenzyme A (PA-CoA) blocked binding. It was concluded that PaaR is the negative regulator of PAA degradation and that PA-CoA is the PaaR effector. In addition, GlxR binding sites were found, and binding to GlxR was confirmed. Therefore, PAA catabolism in C. glutamicum is regulated by the pathway-specific repressor PaaR, and also likely by the global transcription regulator GlxR. By comparative genomic analysis, we reconstructed orthologous PaaR regulons in 57 species, including species of Actinobacteria , Proteobacteria , and Flavobacteria , that carry PAA utilization genes and operate by conserved binding motifs, suggesting that PaaR-like regulation might commonly exist in these bacteria.
机译：近年来，工业上重要的生物体谷氨酸棒杆菌的特征在于其吸收芳族化合物的强大能力。在这项研究中，研究了谷氨酸棒杆菌菌株AS 1.542的分解代谢苯乙酸（PAA）的能力。鉴定了paa基因;它们被组织成连续的paa基因簇。谷氨酸棒杆菌型菌株ATCC 13032不能分解代谢PAA，但是重组菌株ATCC 13032 / pEC-K18 mob 2 :: paa具有在PAA上生长的能力。详细研究了编码TetR家族转录调节子的paaR基因。菌株AS 1.542中paaR的破坏导致所有paa基因的转录增加。确定了转录起始位点和推定的启动子区域。在paa基因的上游区域发现了一个不完善的回文基序（5'-ACTNACCGNNCGNNCGGTNAGT-3'; 22 bp）。电泳迁移率变动分析（EMSA）证明PaaR与该基序具有特异性结合，而苯乙酰辅酶A（PA-CoA）阻止了结合。结论是，PaaR是PAA降解的负调节剂，而PA-CoA是PaaR效应子。另外，发现了GlxR结合位点，并证实了与GlxR的结合。因此，谷氨酸棒杆菌中的PAA分解代谢受途径特异性阻遏物PaaR的调节，也可能受全局转录调节因子GlxR的调节。通过比较基因组分析，我们重建了57种物种的直系同源PaaR调控因子，包括放线菌属，变形杆菌属和黄杆菌属的物种，这些物种携带PAA利用基因并通过保守的结合基序进行操作，这表明这些细菌中可能普遍存在类PaaR调控。

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