Biosynthesis of mannose from glucose via constructing phosphorylation-dephosphorylation reactions in Escherichia coli

IF 3.4 3区生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Enzyme and Microbial Technology Pub Date : 2024-03-18 DOI:10.1016/j.enzmictec.2024.110427

Yuyao Wang , Enhui Chen , Yanfei Wang , Xinming Sun , Qianzhen Dong , Peng Chen , Chenglin Zhang , Jiangang Yang , Yuanxia Sun

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引用次数: 0

Abstract

d-mannose has been widely used in food, medicine, cosmetic, and food-additive industries. To date, chemical synthesis or enzymatic conversion approaches based on iso/epimerization reactions for d-mannose production suffered from low conversion rate due to the reaction equilibrium, necessitating intricate separation processes for obtaining pure products on an industrial scale. To circumvent this challenge, this study showcased a new approach for d-mannose synthesis from glucose through constructing a phosphorylation-dephosphorylation pathway in an engineered strain. Specifically, the gene encoding phosphofructokinase (PfkA) in glycolytic pathway was deleted in Escherichia coli to accumulate fructose-6-phosphate (F6P). Additionally, one endogenous phosphatase, YniC, with high specificity to mannose-6-phosphate, was identified. In ΔpfkA strain, a recombinant synthetic pathway based on mannose-6-phosphate isomerase and YniC was developed to direct F6P to mannose. The resulting strain successfully produced 25.2 g/L mannose from glucose with a high conversion rate of 63% after transformation for 48 h. This performance surpassed the 15% conversion rate observed with 2-epimerases. In conclusion, this study presents an efficient method for achieving high-yield mannose synthesis from cost-effective glucose.

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大肠杆菌通过构建磷酸化-磷酸化反应从葡萄糖中生物合成甘露糖

d-mannose 已广泛应用于食品、医药、化妆品和食品添加剂行业。迄今为止，基于异构/epimerization 反应的化学合成或酶转化方法生产的 d-甘露糖因反应平衡而转化率较低，需要复杂的分离过程才能在工业规模上获得纯产品。为了规避这一挑战，本研究通过在工程菌株中构建磷酸化-磷酸化途径，展示了一种从葡萄糖合成 d-甘露糖的新方法。具体来说，在大肠杆菌中删除了编码糖酵解途径中磷酸果糖激酶（PfkA）的基因，以积累 6-磷酸果糖（F6P）。此外，还发现了一种对 6-磷酸甘露糖具有高度特异性的内源性磷酸酶 YniC。在 ΔpfkA 菌株中，开发出了一种基于 6-磷酸甘露糖异构酶和 YniC 的重组合成途径，可将 F6P 直接转化为甘露糖。转化 48 小时后，所产生的菌株成功地从葡萄糖中生产出 25.2 克/升的甘露糖，转化率高达 63%。这一表现超过了 2-嵌合酶 15%的转化率。总之，本研究提出了一种高效的方法，可从低成本的葡萄糖中获得高产的甘露糖合成。

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来源期刊

Enzyme and Microbial Technology 生物-生物工程与应用微生物

CiteScore

7.60

自引率

5.90%

发文量

142

审稿时长

38 days

期刊介绍： Enzyme and Microbial Technology is an international, peer-reviewed journal publishing original research and reviews, of biotechnological significance and novelty, on basic and applied aspects of the science and technology of processes involving the use of enzymes, micro-organisms, animal cells and plant cells. We especially encourage submissions on: Biocatalysis and the use of Directed Evolution in Synthetic Biology and Biotechnology Biotechnological Production of New Bioactive Molecules, Biomaterials, Biopharmaceuticals, and Biofuels New Imaging Techniques and Biosensors, especially as applicable to Healthcare and Systems Biology New Biotechnological Approaches in Genomics, Proteomics and Metabolomics Metabolic Engineering, Biomolecular Engineering and Nanobiotechnology Manuscripts which report isolation, purification, immobilization or utilization of organisms or enzymes which are already well-described in the literature are not suitable for publication in EMT, unless their primary purpose is to report significant new findings or approaches which are of broad biotechnological importance. Similarly, manuscripts which report optimization studies on well-established processes are inappropriate. EMT does not accept papers dealing with mathematical modeling unless they report significant, new experimental data.