[P006] Poster

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

[P03-312] Engineering Tunable Translational Coupling Bioparts for Multi-Gene Expression Control in Corynebacterium glutamicum

○Eunseo Jeong¹, JISEON LEE¹ (1. Department of Biological Sciences and Biotechnology, Chonnam National University (Korea))

Keywords:

corynebacterium glutamicum,synthetic bioparts,Translational coupling

While Corynebacterium glutamicum is a robust industrial chassis widely used for the production of value-added biochemicals, the scarcity of tunable genetic parts limits the optimization of complex multi-gene biosynthetic pathways. Traditional operons frequently suffer from unpredictable translational efficiencies, resulting in imbalanced protein expression ratios. To address this, we developed predictable translational coupling (TC) bioparts for regulating multi-gene expression, utilizing strategies such as translation-reinitiation and upstream translation-dependent de novo initiation. We first constructed genome-derived sequence-based parts by mining endogenous loci featuring overlapping stop-start codons and inserted them between the mCherry and eGFP reporter genes. Subsequent analysis revealed higher downstream reporter expression for several sequences. This suggests that cryptic internal RBSs may have induced upstream translation-dependent de novo initiation. To precisely control these expression traits, we introduced synthetic sequence-based parts by rationally designing RNA secondary structures within these regions. By tuning the thermodynamic stability (ΔG_unfold), we masked cryptic binding sites and modulated translational read-through. Ultimately, these diverse TC modules allow predictable tuning of downstream expression ratios and could significantly expand the synthetic biology toolkit for C. glutamicum