[P004] Poster

1:00 PM - 1:50 PM JST(4:00 AM - 4:50 AM UTC)

[P02-190] Toward identifying the genetic basis for the adaptation of Escherichia coli NOG Strain

○Angela H. Chao¹, Lusiana Ekawati Angelina¹, Celine Y. Li¹, Paul P. Lin¹ (1. National Yang Ming Chiao Tung University (Taiwan))

Keywords:

Non-oxidative glycolysis (NOG),Adaptive laboratory evolution (ALE),Synthetic biology

The presence of multiple mutations accumulated during evolution makes it challenging to identify key genetic changes. In the previous study, an engineered Escherichia coli strain was designed to rely solely on non-oxidative glycolysis (NOG) pathway for sugar metabolism, eliminates the carbon loss associated with the Embden-Meyerhof-Parnas (EMP) pathway. The strain was constructed through rational design followed by two rounds of adaptive laboratory evolution (ALE). (Lin et al., 2018). Various mutations occurred during ALE, rerouting metabolism and altering its regulatory mechanisms. However, the essential mutations responsible for the observed phenotypic changes remain unknown. In this study, we aim to identify the key mutations during the ALE that contribute to the NOG strain’s evolution. Using a reverse engineering screening strategy, we assess the effects of restoring the normal gene function of each mutation. Our initial results indicate that ptsG, araE, gadW, yiaT, ptsI and narH may play a significant role in the first round of ALE while fadR, rpoS, yhaJ and gluQ may play critical roles in the second round. Further engineering will be conducted to confirm these findings. Identifying these essential mutations will provide valuable insights for further rational engineering, enabling the development of a more robust NOG strain with enhanced metabolic efficiency.