Enzymatic production of HMO mimics by the sialylation of galacto-oligosaccharides.
Enzymatic production of HMO mimics by the sialylation of galacto-oligosaccharides.
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DOI：
10.1016/j.foodchem.2015.02.064
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发表时间：
2015-08
期刊：
影响因子：
8.8
通讯作者：
Yuanming Wang;Kai-Jun Jiang;Huanna Ma;W. Zeng;P. Wang;Nana Yao;Weiqing Han;Jiansong Cheng;Wei Wang
中科院分区：
文献类型：
--
作者：
Yuanming Wang;Kai-Jun Jiang;Huanna Ma;W. Zeng;P. Wang;Nana Yao;Weiqing Han;Jiansong Cheng;Wei Wang
母乳低聚糖 (HMO) 是一类结构多样的非共轭聚糖，具有广泛的生物活性。在本报告中，我们描述了一种高效的多酶一锅法策略，通过低聚半乳糖 (GOS) 的唾液酸化来生产 HMO 模拟物，低聚半乳糖 (GOS) 经常添加到婴儿配方奶粉中，作为 HMO 的廉价替代品。在该系统中，唾液酸转移酶供体胞苷-5'-单磷酸-N-乙酰神经氨酸（CMP-Neu5Ac）是使用 CMP 唾液酸合成酶原位生成的。使用碱性磷酸酶 PhoA 将 CMP 消化为胞苷和无机磷酸盐后，通过一步纯化获得唾液酸化的 GOS。尽管合成的α2,3-、α2,6-和α2,3/8-唾液酸-GOS表现出不同的唾液酸化水平和模式，但所有这些混合物都可以被长双歧杆菌婴儿亚种ATCC 15697发酵，但不能被青春双歧杆菌ATCC 15703发酵。前一菌株特有的唾液酸酶NanH2可以水解所有合成的产物。 HMO 模仿。
Human milk oligosaccharides (HMOs) are a family of structurally diverse unconjugated glycans that exhibit a wide range of biological activities. In this report, we describe an efficient, Multi-Enzyme One-Pot strategy to produce HMO mimics by the sialylation of galacto-oligosaccharides (GOSs), which are often added to infant formula as an inexpensive alternative to HMOs. In this system, the sialyltransferase donor, cytidine-5′-monophospho-N-acetylneuraminic acid (CMP-Neu5Ac), was generatedin situusing a CMP-sialic acid synthetase. The sialylated GOSs were obtained by one-step purification after digesting CMP using the alkaline phosphatase PhoA to cytidine and inorganic phosphate. Although the synthesized α2,3-, α2,6- and α2,3/8-sialyl-GOSs exhibit different sialylation levels and patterns, all of these mixtures can be fermented byBifidobacterium longumsubsp.infantisATCC 15697 but not byBifidobacterium adolescentisATCC 15703. The sialidase NanH2, which is unique to the former strain, hydrolyzed all of the synthesized HMO mimics.