Shewanella oneidensis MR-1 has found widespread applications in pollutant transformation and bioenergy production, closely tied to its outstanding heme synthesis capabilities. However, this significant biosynthetic potential is still unexploited so far. Here, we turned this bacterium into a highly-efficient bio-factory for green synthesis of 5-Aminolevulinic Acid (5-ALA), an important chemical for broad applications in agriculture, medicine, and the food industries. The native C5 pathway genes of S. oneidensis was employed, together with the introduction of foreign anti-oxidation module, to establish the 5-ALA production module, resulting 87-fold higher 5-ALA yield and drastically enhanced tolerance than the wild type. Furthermore, the metabolic flux was regulated by using CRISPR interference and base editing techniques to suppress the competitive pathways to further improve the 5-ALA titer. The engineered strain exhibited 123-fold higher 5-ALA production capability than the wild type. This study not only provides an appealing new route for 5-ALA biosynthesis, but also presents a multi-dimensional modularized engineering strategy to broaden the application scope of S. oneidensis.

中文翻译：

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希瓦氏菌 MR-1 的模块化工程技术用于 5-氨基乙酰丙酸的高效定向合成


希瓦氏菌 MR-1 在污染物转化和生物能源生产中得到了广泛的应用，这与其出色的血红素合成能力密切相关。然而，到目前为止，这种巨大的生物合成潜力仍未得到开发。在这里，我们将这种细菌变成了一种高效的生物工厂，用于绿色合成 5-氨基乙酰丙酸 （5-ALA），5-ALA是一种重要的化学品，在农业、医药和食品工业中具有广泛的应用。采用 S. oneidensis 的天然 C5 通路基因，结合外源抗氧化模块的引入，建立 5-ALA 生产模块，导致 5-ALA 产量比野生型高 87 倍，耐受性显著提高。此外，通过使用 CRISPR 干扰和碱基编辑技术来抑制竞争途径以进一步提高 5-ALA 滴度，从而调节代谢通量。工程菌株的 5-ALA 生产能力比野生型高 123 倍。本研究不仅为 5-ALA 生物合成提供了一条有吸引力的新路线，而且提出了一种多维模块化工程策略，以拓宽 S. oneidensis 的应用范围。