Efficient Biosynthesis of Lacto-N-Biose I, a Building Block of Type I Human Milk Oligosaccharides, by a Metabolically Engineered Escherichia coli,Journal of Agricultural and Food Chemistry

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Efficient Biosynthesis of Lacto-N-Biose I, a Building Block of Type I Human Milk Oligosaccharides, by a Metabolically Engineered Escherichia coli
Journal of Agricultural and Food Chemistry ( IF 5.7 ) Pub Date : 2024-03-07 , DOI: 10.1021/acs.jafc.4c00153
Mengting Tao ₁ , Longhao Yang ₁ , Chunhua Zhao ₂ , Wenli Zhang ₁ , Yingying Zhu ₁ , Wanmeng Mu ₁

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Lacto-N-biose I (LNB), termed a Type 1 disaccharide, is an important building block of human milk oligosaccharides. It shows promising prebiotic activity by stimulating the proliferation of many gut-associated bifidobacteria and thus displays good potential in infant foods or supplements. Enzymatic and microbial approaches to LNB synthesis have been studied, almost all of which involve glycosylation of LNB phosphorylase as the final step. Herein, we report a new and easier microbial LNB synthesis strategy through the route “lactose → lacto-N-triose II (LNTri II) → lacto-N-tetraose (LNT) → LNB”. A previously constructed LNT-producing Escherichia coli BL21(DE3) strain was engineered for LNB biosynthesis by introducing Bifidobacterium bifidum LnbB. LNB was efficiently produced, accompanied by lactose regeneration. Genomic integration of key pathway genes related to LNTri II and LNT synthesis was performed to enhance LNB titers. The final engineered strain produced 3.54 and 26.88 g/L LNB by shake-flask and fed-batch cultivation, respectively.

中文翻译：

代谢工程大肠杆菌高效生物合成 Lacto-N-Biose I（I 型母乳低聚糖的组成部分）

乳-N-二糖 I (LNB)，称为 1 型二糖，是母乳低聚糖的重要组成部分。它通过刺激许多肠道相关双歧杆菌的增殖而显示出有前景的益生元活性，因此在婴儿食品或补充剂中显示出良好的潜力。人们已经研究了 LNB 合成的酶促和微生物方法，几乎所有方法都涉及 LNB 磷酸化酶的糖基化作为最后一步。在此，我们报告了一种新的、更简单的微生物LNB合成策略，通过“乳糖→乳糖-N-三糖II（LNTri II）→乳糖-N-四糖（LNT）→LNB”路线。通过引入两歧双歧杆菌LnbB，对先前构建的产生 LNT 的大肠杆菌BL21(DE3) 菌株进行改造，用于 LNB 生物合成。 LNB 得到有效生产，同时乳糖再生。对与 LNTri II 和 LNT 合成相关的关键途径基因进行基因组整合，以提高 LNB 滴度。最终的工程菌株通过摇瓶和补料分批培养分别产生 3.54 和 26.88 g/L LNB。

更新日期：2024-03-07

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