路径优化和尿苷 5'-三磷酸再生增强工程菌中乳四糖的生物合成。
Pathway Optimization and Uridine 5'-Triphosphate Regeneration for Enhancing Lacto--Tetraose Biosynthesis in Engineered .
机构信息
State Key Laboratory of Food Science and Technology, Jiangnan University, Wuxi, Jiangsu 214122, China.
出版信息
J Agric Food Chem. 2022 Jun 29;70(25):7727-7735. doi: 10.1021/acs.jafc.2c02426. Epub 2022 Jun 19.
Recently, human milk oligosaccharides (HMOs) have attracted increasing attention and display great commercial importance, especially for the infant formula industry. Lacto--tetraose (LNT) is an important neutral HMO commercially added in infant formula and a core structure for synthesizing complex HMOs. Previously, a novel LNT-generating β-1,3-galactosyltransferase from was identified and used for construction of an LNT-producing engineered . In this work, LNT biosynthesis was further enhanced by pathway optimization and uridine 5'-triphosphate (UTP) regeneration. The main strategies included genomic integration of UDP-glucose 4-epimerase-encoding gene, fine-tuning of the LNT pathway-related genes, blocking of competitive pathways related to UDP-galactose, and overexpression of UTP supply related genes. The maximal LNT titer reached 6.16 and 57.5 g/L by shake-flask and fed-batch fermentation, respectively.
最近，人乳寡糖（HMOs）受到越来越多的关注，具有巨大的商业价值，特别是对婴儿配方奶粉行业而言。乳四糖（LNT）是婴儿配方奶粉中添加的一种重要的中性 HMO，也是合成复杂 HMO 的核心结构。先前，从 中鉴定出一种新型的生成 LNT 的β-1,3-半乳糖基转移酶，并将其用于构建 LNT 生产的工程菌。在这项工作中，通过途径优化和尿苷 5'-三磷酸（UTP）再生进一步增强了 LNT 的生物合成。主要策略包括基因组整合 UDP-葡萄糖 4-差向异构酶编码基因、精细调整 LNT 途径相关基因、阻断与 UDP-半乳糖相关的竞争途径以及过表达 UTP 供应相关基因。摇瓶和分批补料发酵的最大 LNT 产量分别达到 6.16 和 57.5 g/L。
Zotero 插件