[2S00a] 01. ASBA - Part 5

2:20 PM - 2:35 PM JST(5:20 AM - 5:35 AM UTC)

[2ASBA-08] Evolution-Guided Microbial Chassis Engineering for Enhanced Bioconversion

○Yong Hee Han¹ (1. School of Biological Sciences and Technology, Chonnam National University (Korea))

Keywords:

Evolutionary engineering,Adaptive laboratory evolution,Biosensor,Tn-seq,Biorefinery

Bioconversion of non-preferred biomass into high-value products requires complex cellular rewiring beyond the mere introduction of heterologous genes, which cannot be fully achieved through rational design alone. We aimed to construct microbial chassis with enhanced biomass uptake and target metabolite production using evolutionary engineering strategies. We first integrated rational design with adaptive laboratory evolution (ALE) to develop a versatile microbial platform capable of efficient carbon utilization and improved biochemical production. ALE was conducted in Vibrio species to enhance the utilization of xylose or raffinose, substrates for which Vibrio exhibits incomplete metabolic pathways or low consumption rates. The evolved strains were further engineered to produce high-value biochemicals, such as lactic acid and citramalate, achieving competitive titers. We also conducted biosensor-guided ALE in Escherichia coli, which redirected carbon flux to balance cell growth and 3-hydroxypropionic acid (3-HP) synthesis, resulting in the highest 3-HP yield. Furthermore, we developed an integrated mutagenesis and genome engineering platform that combines transposon sequencing, biosensor-based selection, and MAGE, which accelerated the evolutionary engineering process and significantly enhanced naringenin production. Collectively, these results demonstrate the power of evolution-guided strategies in constructing microbial chassis with broadened substrate ranges and superior metabolic performance for sustainable biomanufacturing.