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使用多酶级联与辅助因子再生合成2'-糖糖

Chenqi Cao¹, Yi Bai², Mengxin Zhang¹

¹State Key Laboratory of Green Biomanufacturing, College of Life Science and Technology, Beijing University of Chemical Technology, Beijing, 100029, China.

Scientific reports

|December 13, 2025

概括

研究人员开发了一种新的 in vitro 酶系统,用于从曼诺和乳糖中合成2 - 糖 (2′-FL). 这种高效的途径显著减少了辅助因子的使用,为生产这种重要的母乳寡糖化物提供了一种新方法.

科学领域:

生物化学和合成生物学
酶合成酶的合成
碳水化合物化学碳水化合物化学

背景情况:

2′-福科西拉克托斯 (2′-FL) 是一种重要的母乳寡糖,具有显著的健康益处.
现有的2′-FL合成方法主要侧重于从葡萄糖和甘油的体内路径.
需要有效和可扩展的体外合成方法来生产2′-FL.

研究的目的:

建立第一个体外多酶级联系统,用于2′-FL的新合成.
为了利用曼诺和乳糖作为2′-FL生产的基质.
设计和优化酶和辅助因子再生,以提高2′-FL产量.

主要方法:

设计了一种工程性聚酸盐依赖型葡萄糖酶 (PPGMK) 用于曼诺斯酸化.
优化了多酶级联系统,包括GDP-D-mannose-4,6-dehydratase (gmd) 和α-1,2-fucosyltransferase.
实施了一种高效的GTP再生系统,使用聚酸盐激酶2进行辅因子回收.

主要成果:

通过系统优化和GTP再生,实现了2′-FL产量的2.94倍改善.
在使用曼诺斯和乳糖的单反应中合成了13.18mM 2′-FL.
证明了异常的辅助因子效率,减少了NADPH和GTP消耗的6.6倍.

结论:

为2′-FL合成建立了一个新的de novo酶途径.
确定了GDP-D-mannose-4,6-dehydratase作为一个关键的速度限制酶.
提供了对 oligosaccharide 生产路径构建和酶工程的见解.

关键词:

2′-Fucosyllactose 的使用情况.在GTP再生的再生过程中.人类牛奶中的寡糖类.多酶级联是多种酶的级联.一个α-1,2-fucosyl转移酶.

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