Department of Life Sciences and Institute of Genome Sciences, National Yang Ming Chiao Tung University, Taipei 112, Taiwan.
Department of Agricultural Chemistry, National Taiwan University, Taipei 10617, Taiwan.
J Agric Food Chem. 2024 Aug 21;72(33):18649-18657. doi: 10.1021/acs.jafc.4c03661. Epub 2024 Aug 7.
Trehalose synthase (TreS) catalyzes the reversible interconversion of maltose to trehalose, playing a vital role in trehalose production. Understanding the catalytic mechanism of TreS is crucial for optimizing the enzyme activity and enhancing its suitability for industrial applications. Here, we report the crystal structures of both the wild type and the E324D mutant of trehalose synthase in complex with the trehalose analogue, validoxylamine A. By employing structure-guided mutagenesis, we identified N253, E320, and E324 as crucial residues within the +1 subsite for isomerase activity. Based on these complex structures, we propose the catalytic mechanism underlying the reversible interconversion of maltose to trehalose. These findings significantly advance our comprehension of the reaction mechanism of TreS.
海藻糖合酶（TreS）催化麦芽糖向海藻糖的可逆转化，在海藻糖生产中起着至关重要的作用。了解 TreS 的催化机制对于优化酶活性和增强其在工业应用中的适用性至关重要。在这里，我们报告了野生型和 E324D 突变体海藻糖合酶与海藻糖类似物 validoxylamine A 复合物的晶体结构。通过结构指导的诱变，我们确定了 N253、E320 和 E324 是异构酶活性的 +1 亚位点中的关键残基。基于这些复合物结构，我们提出了麦芽糖向海藻糖可逆转化的催化机制。这些发现极大地促进了我们对 TreS 反应机制的理解。
