[3S00a] 01. ASBA - Part 8

2:50 PM - 3:05 PM JST(5:50 AM - 6:05 AM UTC)

[3ASBA-11] Using Bacteria as Living Test Tubes to Study Human Gene Mutations and Find New Drug Leads

○Donghui Choe¹, Bernhard Palsson² (1. Sungkyunkwan University (Korea), 2. University of California San Diego (USA))

Keywords:

Synthetic Biology,Biomedical Engineering,Living Test Tube

Recent advances in high-throughput sequencing have uncovered countless genetic variations linked to human diseases. However, clarifying their functional significance remains a major challenge. We present an alternative approach that employs humanized Escherichia coli to study human genetic enzymopathies and evaluate candidate drug effects on metabolic enzymes. By replacing selected endogenous E. coli metabolic enzymes with their human orthologs and their sequence variants, we demonstrate that the growth rate of E. coli reflects the in vivo activity of heterologously expressed human enzymes. The assay accurately distinguished pathogenic from benign variants of glucose-6-phosphate isomerase (GPI) and glucose-6-phosphate dehydrogenase (G6PD) and enabled rapid identification of less-characterized sequence variations associated with human diseases. We further expanded the assay to argininosuccinate lyase (ASL), demonstrating its versatility in capturing deficiencies in enzymes from diverse metabolic pathways. Moreover, the use of humanized E. coli offers a cost-efficient and scalable strategy for drug discovery. The assay successfully reproduced the effects of known G6PD inhibitors and activators and enabled rapid screening of a 160-compound library. From this library, seven lead compounds were identified, comprising both previously known inhibitors and four newly discovered inhibitors with strong potency against human G6PD. Together, these findings establish the assay as a scalable, cost-efficient platform that bridges the gap between in vitro biochemical assays and cell-based systems. Continued research and innovation in this field have the potential to impact the development and practice of precision medicine.