Combinatorial engineering of microbial cell factory for 2′-fucosyllactose biosynthesis

IF 3.7 3区生物学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY

Process Biochemistry Pub Date : 2025-06-04 DOI:10.1016/j.procbio.2025.05.024

Jingyu Zhou , Xinyue Su , Yongzhong Wang , Qiang Ding

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引用次数: 0

Abstract

2’-fucosyllactose (2’-FL), a predominant human milk oligosaccharide (HMOs), has showed significant potential in infant health applications including intestinal protection, cognitive development, and immune system modulation. Therefore, to enhance microbial biosynthesis of 2’-FL, we engineered a de novo synthesis pathway through deleting key enzymes gene: wecB (encoding UDP-N-acetyl glucosamine-2-epimerase), lacZ (encoding β-galactosidase), wcaJ (encoding UDP-glucose lipid carrier transferase), which could increase the 2’-FL titer and productivity to 2.25 g/L and 0.03 g/L/h, respectively. Furthermore, optimization of induction time further improved production performance to 4.06 g/L and 0.06 g/L/h. Through systematic gene deletion analysis, we identified lacA and iclR as critical metabolic nodes. Their knockout negatively impacted 2’-FL biosynthesis by reducing lactose availability and disturbing metabolic flux. Finally, the 2’-FL was furthermore improved to 4.75 g/L through expressing gsk and zwf genes for NADPH and GTP regeneration. This study can effectively increase the production of 2’-fucosyllactose, which can provide a research basis for the large-scale bioproduction of 2’-fucosyllactose in the industrial factory.

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2′-焦酰基乳糖生物合成微生物细胞工厂的组合工程

2 ' -焦酰基乳糖（2 ' - fl）是一种主要的母乳低聚糖（HMOs），在婴儿健康方面具有重要的应用潜力，包括肠道保护、认知发展和免疫系统调节。因此，为了提高微生物对2′-FL的合成，我们通过删除关键酶基因wecB（编码udp - n -乙酰氨基葡萄糖-2- epimase）、lacZ（编码β-半乳糖苷酶）、wcaJ（编码udp -葡萄糖脂质载体转移酶），设计了一条新的合成途径，使2′-FL的滴度和产率分别提高到2.25 g/L和0.03 g/L/h。优化诱导时间可进一步提高生产性能，分别达到4.06 g/L和0.06 g/L/h。通过系统的基因缺失分析，我们发现lacA和iclR是关键的代谢节点。它们的敲除通过降低乳糖可利用性和扰乱代谢通量对2 ' -FL生物合成产生负面影响。最后，通过表达gsk和zwf基因使NADPH和GTP再生，2′-FL进一步提高到4.75 g/L。本研究可有效提高2′-焦糖乳糖的产量，为工业化工厂大规模生物生产2′-焦糖乳糖提供研究依据。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Process Biochemistry 生物-工程：化工

CiteScore

8.30

自引率

4.50%

发文量

374

审稿时长

53 days

期刊介绍： Process Biochemistry is an application-orientated research journal devoted to reporting advances with originality and novelty, in the science and technology of the processes involving bioactive molecules and living organisms. These processes concern the production of useful metabolites or materials, or the removal of toxic compounds using tools and methods of current biology and engineering. Its main areas of interest include novel bioprocesses and enabling technologies (such as nanobiotechnology, tissue engineering, directed evolution, metabolic engineering, systems biology, and synthetic biology) applicable in food (nutraceutical), healthcare (medical, pharmaceutical, cosmetic), energy (biofuels), environmental, and biorefinery industries and their underlying biological and engineering principles.