Substrate and Process Engineering for Biocatalytic Synthesis and Facile Purification of Human Milk Oligosaccharides.
Substrate and Process Engineering for Biocatalytic Synthesis and Facile Purification of Human Milk Oligosaccharides.
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DOI：
10.1002/cssc.202102539
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发表时间：
2022-05-06
期刊：
影响因子：
8.4
通讯作者：
Chen, Xi
中科院分区：
文献类型：
--
作者：
Bai, Yuanyuan;Yang, Xiaohong;Yu, Hai;Chen, Xi
工艺开发方面的创新对于将生物催化应用于有机化合物的工业和实验室生产至关重要，这些有机化合物包括有益的碳水化合物，如人乳低聚糖(HMOS)。HMO因探索其作为关键成分在改善人类健康产品中的潜在应用而受到越来越多的关注。为了通过生物催化有效地获取HMO，我们开发了一种结合底物和过程工程的策略，即多步骤一锅多酶(MSOPME)设计。该策略允许在单个反应器中使用单个C18柱纯化过程来获得纯标记的HMO，而不考虑靶子的长度。高产率(71-91%)一锅法合成了20个标记的HMO(83-155 mg)，包括含或不含岩藻糖化或唾液酸化的长链低聚糖，从乳糖苷到非糖基，而无需分离中间低聚糖。克级合成了一种重要的HMO衍生物，标记的LNFP-I(LNFP-I)，产率为84%。标签的去除效率很高(94-97%)，不需要柱净化来产生所需的具有自由还原端的天然HMO。我们设想，这种方法可以很容易地适应大规模的合成和自动化，以允许快速获得HMO，其他多糖和糖共轭化合物。有牛奶和糖吗？通过结合生物催化底物和过程工程的概念，开发了一种高效的多步一锅多酶(MSOPME)合成碳水化合物的策略。单一的C18柱纯化过程对任何目标都是足够的。该策略被证明用于制备和克级合成大量的人乳寡糖(HMOS)，这些低聚糖是工业生物催化生产的有吸引力的目标。
Innovation in process development is essential for applying biocatalysis in industrial and laboratory productions of organic compounds including beneficial carbohydrates such as human milk oligosaccharides (HMOs). HMOs have attracted increasing attention for exploring their potential applications as key ingredients in products that can improve human health. To access HMOs via biocatalysis in an efficient manner, we have developed a combined substrate and process engineering strategy namely the multistep one-pot multienzyme (MSOPME) design. The strategy allows the access to a pure tagged HMO in a single reactor with a single C18-cartridge purification process despite the length of the target. Its efficiency was demonstrated in high-yield (71–91%) one-pot synthesis of twenty tagged HMOs (83–155 mg) including long-chain oligosaccharides with or without fucosylation or sialylation up to nonaoses from a lactoside without the isolation of the intermediate oligosaccharides. Gram-scale synthesis of an important HMO derivative, a tagged lacto-N-fucopentaose-I (LNFP-I), succeeded in 84% yield. Tag removal was achieved in high efficiency (94–97%) without the need for column purification to produce the desired natural HMOs with a free reducing end. We envision that the method can be readily adapted for large-scale synthesis and automation to allow quick access to HMOs, other glycans, and glycoconjugates. Got milk sugar? A highly efficient multistep one-pot multienzyme (MSOPME) strategy is developed for carbohydrate synthesis by integrating biocatalysis substrate and process engineering concepts. A single C18-cartridge purification process is sufficient for any target. The strategy is demonstrated for preparative and gram-scale synthesis of numerous human milk oligosaccharides (HMOs), which are attractive targets for industrial biocatalytic production.
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