堺伊甸菌中参与 PET 降解的 MHETase 和 TPA 降解基因的表达调控。

Yuya Tanaka, Kazumi Hiraga, Masayuki Inui
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摘要

Ideonella sakaiensis 是一种能够降解和消耗聚对苯二甲酸乙二醇酯(PET)的细菌,这种塑料材料以前被认为是不可生物降解的。PET 的降解需要两种酶,即聚对苯二甲酸乙二醇酯水解酶(PETase)和对苯二甲酸单(2-羟乙基)酯水解酶(MHETase),它们能将 PET 分解成对苯二甲酸乙二醇酯(TPA)和乙二醇(EG),作为细菌的碳源。以往的研究主要集中在 PETase 和 MHETase 的酶特性、结构和作用机制上。然而,尚未研究 PETase 和 MHETase 基因表达的调控。本研究发现了一种与 MHETase 启动子 DNA 结合的蛋白质,即 I. sakaiensis 中的 MHETase 基因调控蛋白(MRP)。PET 或 TPA 可诱导 PETase 和 MHETase 基因的表达。此外,通过删除 mrp 基因,MHETase 基因的诱导作用被取消,而 PETase 基因的表达却得以保持。此外,参与 TPA 代谢的基因在 mrp 突变体中没有被诱导。此外,PET 和 TPA 的生长因 mrp 突变而恶化。此外,MRP 与 MHETase 基因和 TPA 代谢基因的启动子区域结合,但与 PETase 基因启动子没有结合。这些结果表明,MRP 是一种能激活 MHETase 和 TPA 代谢基因的转录因子。
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Regulation of the expression of MHETase and TPA degradation genes involved in the degradation of PET in Ideonella sakaiensis.

Ideonella sakaiensis is a bacterium that can degrade and consume polyethylene terephthalate (PET), a plastic material that was previously considered non-biodegradable. The degradation of PET requires two enzymes, namely poly (ethylene terephthalate) hydrolase (PETase) and mono (2-hydroxyethyl) terephthalate hydrolase (MHETase), which break down PET into terephthalate (TPA) and ethylene glycol (EG), which serve as carbon sources for the bacterium. Previous studies have focused on the enzymatic properties, structure, and mechanism of action of PETase and MHETase. However, the regulation of PETase and MHETase gene expression has not been investigated. This study identified a protein that binds to the MHETase promoter DNA, MHETase gene-regulating protein (MRP) in I. sakaiensis. PET or TPA induced the expression of PETase and MHETase genes. Furthermore, the induction of the MHETase gene was abolished by the deletion of the mrp gene, while the expression of the PETase gene was maintained. In addition, the genes involved in TPA metabolism were not induced in the mrp mutant. Furthermore, the growth of the PET and TPA deteriorated due to mrp mutation. Also, MRP binds to the promoter regions of the MHETase gene and TPA metabolizing genes, but not to the PETase gene promoter. These results suggest that MRP is a transcription factor that activates MHETase and TPA-metabolizing genes.

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