Dipicolinic acid (DPA), a cyclic diacid, has garnered significant interest due to its potential applications in antimicrobial agents, antioxidants, chelating reagents, and polymer precursors. However, its natural bioproduction is limited since DPA is only accumulated in Bacillus and Clostridium species during sporulation. Thus, heterologous production by engineered strains is of paramount importance for developing a sustainable biological route for DPA production. Pseudomonas putida KT2440 has emerged as a promising host for the production of various chemicals thanks to its robustness, metabolic versatility, and genetic tractability. The dominant Entner–Doudoroff (ED) pathway for glucose metabolism in this strain offers an ideal route for DPA production due to the advantage of NADPH generation and the naturally balanced flux between glyceraldehyde-3-phosphate and pyruvate, which are both precursors for DPA synthesis. In this study, DPA production via the ED pathway was in silico designed in P. putida KT2440. The systematically engineered strain produced dipicolinate with a titer of 11.72 g/L from glucose in a 5 L fermentor. This approach not only provides a sustainable green route for DPA production but also expands our understanding of the metabolic potential of the ED pathway in P. putida KT2440.

中文翻译：

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工程恶臭假单胞菌 KT2440 通过 Entner-Doudoroff 途径生产吡啶二羧酸

吡啶二羧酸 (DPA) 是一种环状二酸，由于其在抗菌剂、抗氧化剂、螯合剂和聚合物前体方面的潜在应用而引起了人们的极大兴趣。然而，其自然生物生产是有限的，因为 DPA 仅在芽孢杆菌和梭菌属物种中在孢子形成过程中积累。因此，工程菌株的异源生产对于开发 DPA 生产的可持续生物途径至关重要。由于其稳健性、代谢多样性和遗传易处理性，恶臭假单胞菌KT2440 已成为生产各种化学品的有前途的宿主。该菌株中葡萄糖代谢的主导 Entner-Doudoroff (ED) 途径为 DPA 生产提供了理想的途径，因为 NADPH 生成的优势以及 3-磷酸甘油醛和丙酮酸之间的自然平衡通量（两者都是 DPA 合成的前体） 。在这项研究中，通过 ED 途径生产 DPA 是在恶臭假单胞菌KT2440 中进行的计算机设计。该系统工程菌株在 5 L 发酵罐中从葡萄糖中产生效价为 11.72 g/L 的吡啶二羧酸。该方法不仅为 DPA 生产提供了一条可持续的绿色路线，而且扩大了我们对恶臭假单胞菌KT2440 中 ED 途径代谢潜力的理解。