Donor substrate promiscuity of bacterial β1-3-N-acetylglucosaminyltransferases and acceptor substrate flexibility of β1-4-galactosyltransferases.
Donor substrate promiscuity of bacterial β1-3-N-acetylglucosaminyltransferases and acceptor substrate flexibility of β1-4-galactosyltransferases.
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DOI：
10.1016/j.bmc.2016.02.043
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发表时间：
2016-04-15
影响因子：
3.5
通讯作者：
Chen X
中科院分区：
文献类型：
--
作者：
Li Y;Xue M;Sheng X;Yu H;Zeng J;Thon V;Chen Y;Muthana MM;Wang PG;Chen X
β1–3-N-乙酰氨基葡萄糖转移酶 (β3GlcNAcTs) 和 β1–4-半乳糖基转移酶 (β4GalTs) 已广泛用于酶促合成含 N-乙酰基乳糖胺 (LacNAc) 的寡糖和糖复合物，包括人乳中发现的聚 LacNAc 和乳糖 N-新四糖 (LNnT)和其他哺乳动物。为了探索可以通过β3GlcNAcTs和β4GalTs组合合成的寡糖和衍生物，使用39个糖核苷酸的文库分别对来自幽门螺杆菌（Hpβ3GlcNAcT）和脑膜炎奈瑟氏菌（NmLgtA）的两种细菌β3GlcNAcTs进行供体底物特异性研究。两个 β3GlcNAcT 具有互补的供体底物混杂性，并产生了 13 种不同的三糖。它们分别用于研究来自脑膜炎奈瑟菌 (NmLgtB)、幽门螺杆菌 (Hpβ4GalT) 和牛 (Bβ4GalT) 的三种 β4GalT 的受体底物特异性。 13 种三糖中有 10 种被证明是这些 β4GalT 中至少一种的可耐受受体。演示了 NmLgtA 在一锅多酶（OPME）合成两种三糖（包括 GalNAcβ1–3Galβ1–4GlcβProN3 和 Galβ1–3Galβ1–4Glc）中的应用。该研究为使用这些糖基转移酶作为寡糖及其衍生物的酶促和化学酶合成中的强大催化剂提供了重要信息，这些寡糖及其衍生物可作为有用的探针和试剂。
β1–3-N-Acetylglucosaminyltransferases (β3GlcNAcTs) and β1–4-galactosyltransferases (β4GalTs) have been broadly used in enzymatic synthesis of N-acetyllactosamine (LacNAc)-containing oligosaccharides and glycoconjugates including poly-LacNAc, and lacto-N-neotetraose (LNnT) found in the milk of human and other mammals. In order to explore oligosaccharides and derivatives that can be synthesized by the combination of β3GlcNAcTs and β4GalTs, donor substrate specificity studies of two bacterial β3GlcNAcTs from Helicobacter pylori (Hpβ3GlcNAcT) and Neisseria meningitidis (NmLgtA), respectively, using a library of 39 sugar nucleotides were carried out. The two β3GlcNAcTs have complementary donor substrate promiscuity and 13 different trisaccharides were produced. They were used to investigate the acceptor substrate specificities of three β4GalTs from Neisseria meningitidis (NmLgtB), Helicobacter pylori (Hpβ4GalT), and bovine (Bβ4GalT), respectively. Ten of the 13 trisaccharides were shown to be tolerable acceptors for at least one of these β4GalTs. The application of NmLgtA in one-pot multienzyme (OPME) synthesis of two trisaccharides including GalNAcβ1–3Galβ1–4GlcβProN3 and Galβ1–3Galβ1–4Glc was demonstrated. The study provides important information for using these glycosyltransferases as powerful catalysts in enzymatic and chemoenzymatic syntheses of oligosaccharides and derivatives which can be useful probes and reagents.