Crystal Structure and Functional Characterization of the Bifunctional N-(5′-Phosphoribosyl)anthranilate Isomerase-indole-3-glycerol-phosphate Synthase from Corynebacterium glutamicum,Journal of Agricultural and Food Chemistry

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Crystal Structure and Functional Characterization of the Bifunctional N-(5′-Phosphoribosyl)anthranilate Isomerase-indole-3-glycerol-phosphate Synthase from Corynebacterium glutamicum
Journal of Agricultural and Food Chemistry ( IF 5.7 ) Pub Date : 2021-10-16 , DOI: 10.1021/acs.jafc.1c05132
Woojin Park ₁ , Hyeoncheol Francis Son ₁ , Donghoon Lee ₁ , Il-Kwon Kim ₁ , Kyung-Jin Kim ₁

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L-Tryptophan is known as an aromatic amino acid and one of the essential amino acids that must be ingested through various additives or food. TrpCF is a bifunctional enzyme that has indole-glycerol-phosphate synthase (IGPS) and phosphoribosylanthranilate isomerase (PRAI) activity. In this report, we identified the crystal structure of TrpCF from Corynebacterium glutamicum (CgTrpCF) and successfully elucidated the active site by attaching rCdRP similar to the substrate and product of the TrpCF reaction. Also, we revealed that CgTrpCF shows a conformational change at the loops upon substrate binding. We analyzed amino acid sequences of the homologues of CgTrpCF, and the residues of the substrate-binding site in TrpCF were highly conserved except for some residues. These less conserved residues were replaced by site-directed mutagenesis experiments. Consequently, we obtained the CgTrpCF^P294K (PRAI^CD/P294K) variant that has enhanced activity.

中文翻译：

谷氨酸棒杆菌双功能 N-(5'-磷酸核糖基)邻氨基苯甲酸异构酶-吲哚-3-甘油-磷酸合酶的晶体结构和功能表征

L-色氨酸被称为芳香族氨基酸，是必须通过各种添加剂或食物摄取的必需氨基酸之一。TrpCF 是一种双功能酶，具有吲哚-甘油-磷酸合酶 (IGPS) 和磷酸核糖邻氨基苯甲酸异构酶 (PRAI) 活性。在本报告中，我们从谷氨酸棒状杆菌( Cg TrpCF) 中鉴定了TrpCF 的晶体结构，并通过将类似于 TrpCF 反应的底物和产物的 rCdRP 连接起来，成功阐明了活性位点。此外，我们发现Cg TrpCF 在底物结合后在环处显示出构象变化。我们分析了Cg同源物的氨基酸序列除了一些残基外，TrpCF 和 TrpCF 中底物结合位点的残基高度保守。这些不太保守的残基被定点诱变实验取代。因此，我们获得了活性增强的Cg TrpCF ^P294K (PRAI ^CD/P294K ) 变体。

更新日期：2021-10-27

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