Biosynthesis of GDP-D-glycero-α-D-manno-heptose for the Capsular Polysaccharide of Campylobacter jejuni
Biosynthesis of GDP-D-glycero-α-D-manno-heptose for the Capsular Polysaccharide of Campylobacter jejuni
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DOI：
10.1021/acs.biochem.9b00548
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发表时间：
2019-09-17
期刊：
影响因子：
2.9
通讯作者：
Raushel, Frank M.
中科院分区：
文献类型：
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作者：
Huddleston, Jamison P.;Raushel, Frank M.
空肠弯曲菌的荚膜多糖（CPS）结构有助于其稳健的适应性。许多菌株在其CPS单元中含有庚糖部分。前体庚糖是GDP-D-甘油-α-D-甘露-庚糖;在C3-C6处的立体化学修饰以及甲基和氨基磷酸酯基团的添加导致C.空肠CPS结构。先前已经描述了GDP-D-甘油-α-D-甘露-庚糖的合成，但是使用来自嗜热需氧动脉瘤芽孢杆菌DSM 10155的酶。在这里，我们描述了GDP-D-甘油-α-D-甘露庚糖的完全合成使用的酶从C。空肠NTCC 11168：Cj 1152和Cj 1423-Cj 1425。我们的研究结果产生这些酶的动力学参数，并概述了一个成功的策略，毫克-克规模的GDP-D-甘油-α-D-甘露庚糖的合成。这一成就对于其他碳水化合物剪裁酶的表征是至关重要的，这些碳水化合物剪裁酶预期利用GDP-D-甘油-α-D-甘露庚糖用于C.空肠。
The capsular polysaccharide (CPS) structure of Campylobacter jejuni contributes to its robust fitness. Many strains contain heptose moieties in their CPS units. The precursor heptose is GDP-D-glycero-alpha-D-manno-heptose; modifications to the stereochemistry at C3-C6 as well as additions of methyl and phosphoramidate groups lend to the hyper-variability of the C. jejuni CPS structures. Synthesis of GDP-D-glycero-alpha-D-manno-heptose has been described previously, but using enzymes from Aneurinibacillus thermoaerophilus DSM 10155. Here we describe the complete synthesis of GDP-D-glycero-alpha-D-manno-heptose using enzymes from C. jejuni NTCC 11168: Cj1152 and Cj1423-Cj1425. Our results yield kinetic parameters for these enzymes and outline a successful strategy for milligram-gram scale synthesis of GDP-D-glycero-alpha-D-manno-heptose. This achievement is critical for the characterization of other carbohydrate tailoring enzymes, which are expected to utilize GDP-D-glycero-alpha-D-manno-heptose for the biosynthesis of more complex carbohydrates in the CPS of C. jejuni.