School of Energy & Chemical Engineering, Ulsan National Institute of Science and Technology (UNIST), Ulsan, Republic of Korea.
Department of Biomedical Engineering, Ulsan National Institute of Science and Technology (UNIST), Ulsan, Republic of Korea.
Biotechnol J. 2022 Mar;17(3):e2000416. doi: 10.1002/biot.202000416. Epub 2021 May 28.
In this study, whole-cell biotransformation was conducted to produce nonanedioic acid from nonanoic acid by expressing the alkane hydroxylating system (AlkBGT) from Pseudomonas putida GPo1 in Escherichia coli. Following adaptive laboratory evolution, an efficient E. coli mutant strain, designated as MRE, was successfully obtained, demonstrating the fastest growth (27-fold higher) on nonanoic acid as the sole carbon source compared to the wild-type strain. Additionally, the MRE strain was engineered to block nonanoic acid degradation by deleting fadE. The resulting strain exhibited a 12.8-fold increase in nonanedioic acid production compared to the wild-type strain. Six mutations in acrR, P , dppA, P , e14, and yeaR were identified in the mutant MRE strain, which was characterized using genomic modifications and RNA-sequencing. The acquired mutations were found to be beneficial for rapid growth and nonanedioic acid production.
在这项研究中，通过在大肠杆菌中表达来自恶臭假单胞菌 GPo1 的烷烃羟化系统 (AlkBGT)，进行了全细胞生物转化，以从壬酸生产壬二酸。经过适应性实验室进化，成功获得了一种高效的大肠杆菌突变株，命名为 MRE，与野生型菌株相比，其在壬酸作为唯一碳源上的生长速度最快（提高了 27 倍）。此外，通过删除 fadE 来阻断壬酸的降解，对 MRE 菌株进行了工程改造。与野生型菌株相比，该菌株的壬二酸产量增加了 12.8 倍。在突变株 MRE 中鉴定出了 acrR、P、dppA、P、e14 和 yeaR 中的 6 个突变，通过基因组修饰和 RNA 测序对其进行了表征。发现获得的突变有利于快速生长和壬二酸的生产。
