Size-Controlled Chemoenzymatic Synthesis of Homogeneous Oligosaccharides of Neisseria meningitidis W Capsular Polysaccharide.
Size-Controlled Chemoenzymatic Synthesis of Homogeneous Oligosaccharides of Neisseria meningitidis W Capsular Polysaccharide.
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DOI：
10.1021/acscatal.9b05597
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发表时间：
2020-02-21
期刊：
影响因子：
12.9
通讯作者：
Chen X
中科院分区：
文献类型：
--
作者：
Li R;Yu H;Muthana SM;Freedberg DI;Chen X
脑膜炎奈瑟菌（Nm）血清群W （NmW）是导致大多数侵袭性脑膜炎球菌病（IMD）的六种脑膜炎球菌血清群之一。其荚膜多糖（CPS）是一种毒力因子，是NmW CPS-蛋白结合疫苗的关键成分。目前临床使用的NmW CPS-蛋白结合疫苗是有效的，但成本高，产品在CPS和结合水平上都是异质的。为了开发可能更好的NmW CPS疫苗，本文报道了在顺序一锅多酶（OPME）平台上使用多糖合成酶NmSiaDW以粒径控制的方式合成NmW CPS的均质低聚糖。利用获得的结构明确的合成寡糖，用疏水发色团标记，实现了NmSiaDW的详细生化表征。而NmSiaDW的半乳糖基转移酶活性的催化效率随着唾液苷受体底物大小的增加而显著增加，而半乳糖苷受体底物大小的差异对NmSiaDW唾液基转移酶活性的影响不显著。研究发现，供体和受体底物浓度的比例，而不是受体底物的大小，是产生寡糖大小的主要决定因素。已观察到聚合度（DP）大于65的NmW CPS低聚糖。该研究提供了对NmSiaDW荚膜多糖合酶的更好理解，并展示了一个有效的化学酶合成平台，以尺寸控制的方式获得结构明确的NmW CPS低聚糖。
Neisseria meningitidis (Nm) serogroup W (NmW) is one of the six meningococcal serogroups that cause majority of invasive meningococcal diseases (IMD). Its capsular polysaccharide (CPS) is a virulence factor and is a key component in NmW CPS-protein conjugate vaccines. The current clinically used NmW CPS-protein conjugate vaccines are effective but the costs are high and the products are heterogeneous at both the CPS and the conjugate levels. Towards the development of potentially better NmW CPS vaccines, herein we report the synthesis of homogeneous oligosaccharides of NmW CPS in a size-controlled manner using polysaccharide synthase NmSiaDW in a sequential one-pot multienzyme (OPME) platform. Taking advantage of the obtained structurally defined synthetic oligosaccharides tagged with a hydrophobic chromophore, detailed biochemical characterization of NmSiaDW has been achieved. While the catalytic efficiency of the galactosyltransferase activity of NmSiaDW increases dramatically with the increase of the sialoside acceptor substrate size, the size difference of the galactoside acceptor substrate does not influence NmSiaDW sialyltransferase activity significantly. The ratio of donor and acceptor substrate concentrations, but not the size of the acceptor substrates, has been found to be the major determining factor for the sizes of the oligosaccharides produced. NmW CPS oligosaccharides with a degree of polymerization (DP) higher than 65 have been observed. The study provides a better understanding of NmSiaDW capsular polysaccharide synthase and showcases an efficient chemoenzymatic synthetic platform for obtaining structurally defined NmW CPS oligosaccharides in a size-controlled manner.
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影响因子：
4
作者：
Chen, Xi;Varki, Ajit
通讯作者：
Varki, Ajit
DOI：
10.1073/pnas.1800853116
发表时间：
2019-02-05
影响因子：
11.1
作者：
Geissner, Andreas;Reinhardt, Anika;Seeberger, Peter H.
通讯作者：
Seeberger, Peter H.
影响因子：
6.4
作者：
McNamara, Lucy A.;Thomas, Jennifer Dolan;Acosta, Anna
通讯作者：
Acosta, Anna
DOI：
10.1007/978-1-61779-346-2_1
发表时间：
2012
期刊：
Methods in molecular biology (Clifton, N.J.)
影响因子：
--
作者：
Rouphael, Nadine G;Stephens, David S
通讯作者：
Stephens, David S
影响因子：
3.6
作者：
Claus, Heike;Stummeyer, Katharina;Vogel, Ulrich
通讯作者：
Vogel, Ulrich