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Structural Insights into the Catalytic Cycle of Inulin Fructotransferase: From Substrate Anchoring to Product Releasing

文献类型： 外文期刊

作者： Cheng, Mei 1 ; Hou, Xiaodong 3 ; Huang, Zhaolin 1 ; Chen, Ziwei 1 ; Ni, Dawei 1 ; Zhang, Wenli 1 ; Rao, Yijian 3 ; Mu, Wanmeng 1 ;

作者机构： 1.Jiangnan Univ, State Key Lab Food Sci & Technol, Wuxi 214122, Jiangsu, Peoples R China

2.Jiangsu Acad Agr Sci, Inst Agr Prod Proc, Nanjing 210014, Peoples R China

3.Jiangnan Univ, Sch Biotechnol, Key Lab Carbohydrate Chem & Biotechnol, Minist Educ, Wuxi 214122, Jiangsu, Peoples R China

4.Jiangsu Univ, Sch Food & Biol Engn, Zhenjiang 212013, Jiangsu, Peoples R China

关键词： inulin fructotransferase; difructose anhydride; catalytic cycle; substratetrajectory; productpath

期刊名称：JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY （ 影响因子：5.7； 五年影响因子：6.0 ）

ISSN： 0021-8561

年卷期： 2024 年 72 卷 30 期

页码：

收录情况： SCI

摘要： 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.

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