Structural Insights into the Catalytic Cycle of Inulin Fructotransferase: From Substrate Anchoring to Product Releasing,Journal of Agricultural and Food Chemistry

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Structural Insights into the Catalytic Cycle of Inulin Fructotransferase: From Substrate Anchoring to Product Releasing
Journal of Agricultural and Food Chemistry ( IF 5.7 ) Pub Date : 2024-07-22 , DOI: 10.1021/acs.jafc.4c03615
Mei Cheng _{1,

2} , Xiaodong Hou ₃ , Zhaolin Huang ₁ , Ziwei Chen _{1,

4} , Dawei Ni ₁ , Wenli Zhang ₁ , Yijian Rao ₃ , Wanmeng Mu ₁

Affiliation

Carbohydrate degradation is crucial for living organisms due to their essential functions in providing energy and composing various metabolic pathways. Nevertheless, in the catalytic cycle of polysaccharide degradation, the details of how the substrates bind and how the products release need more case studies. Here, we choose an inulin fructotransferase (SpIFTase) as a model system, which can degrade inulin into functionally difructose anhydride I. At first, the crystal structures of SpIFTase in the absence of carbohydrates and complex with fructosyl-nystose (GF4), difructose anhydride I, and fructose are obtained, giving the substrate trajectory and product path of SpIFTase, which are further supported by steered molecular dynamics simulations (MDSs) along with mutagenesis. Furthermore, structural topology variations at the active centers of inulin fructotransferases are suggested as the structural base for product release, subsequently proven by substitution mutagenesis and MDSs. Therefore, this study provides a case in point for a deep understanding of the catalytic cycle with substrate trajectory and product path.

中文翻译：

菊粉果糖转移酶催化循环的结构见解：从底物锚定到产物释放

碳水化合物的降解对于生物体至关重要，因为它们在提供能量和组成各种代谢途径方面发挥着重要作用。然而，在多糖降解的催化循环中，底物如何结合以及产物如何释放的细节需要更多的案例研究。在这里，我们选择菊粉果糖转移酶（ Sp IFTase）作为模型系统，它可以将菊粉降解为功能性二果糖酐I。首先，在没有碳水化合物的情况下， Sp IFTase与果糖基制糖（GF4）复合的晶体结构获得了二果糖酐 I 和果糖，给出了Sp IFTase 的底物轨迹和产物路径，这得到了引导分子动力学模拟 (MDS) 和诱变的进一步支持。此外，菊粉果糖转移酶活性中心的结构拓扑变化被认为是产品释放的结构基础，随后通过替代诱变和 MDS 得到证明。因此，本研究为深入理解底物轨迹和产物路径的催化循环提供了一个恰当的案例。

更新日期：2024-07-22

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