5′-End Translationalization: Iterative Assembly of Leaderless Polycistronic Amplifiers for Context-Independent Expression in the Food-Grade Bacterium <i>Corynebacterium glutamicum</i>

is a crucial food-grade (GRAS) bacterial chassis widely utilized for the industrial production of amino acids and nutraceuticals. However, the efficient production of recombinant proteins and secondary metabolites in this host remains limited by context dependence and low translational efficiency. To overcome this, we introduce a 5'-end translationalization strategy. By repurposing passive 5' untranslated regions (5'UTRs) into actively translated fore-cistrons, we converted conventional monocistronic designs into context-independent, leaderless polycistronic designs (PCDs). This assembly of concatenated fore-cistrons functions as a translational amplifier, largely decoupling protein output from mRNA abundance. We validated this platform by optimizing two biomanufacturing paradigms: achieving a 4.07-fold enhanced secretion of OmlA, a porcine vaccine antigen, and boosting biosynthesis of the food-grade pigment indigoidine to 1.20 g/L (a 7.33-fold increase over baselines). Together, this framework establishes a versatile, portable toolkit to overcome translational bottlenecks, enabling robust hyperproduction of recombinant proteins and engineered metabolites in biotechnology.

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Publication Details

Journal
Journal of Agricultural and Food Chemistry
Published
2026-06-17
DOI
https://doi.org/10.1021/acs.jafc.6c05362
Primary Topic
Microbial Metabolic Engineering and Bioproduction
Type
article
Field-Weighted Citation Impact
0.00

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article

5′-End Translationalization: Iterative Assembly of Leaderless Polycistronic Amplifiers for Context-Independent Expression in the Food-Grade Bacterium Corynebacterium glutamicum

Y ZHAO, Manman Sun, Rodrigo Ledesma‐Amaro, Bin Ye et al.
Journal of Agricultural and Food Chemistry
Microbial Metabolic Engineering and Bioproduction
article

5′-End Translationalization: Iterative Assembly of Leaderless Polycistronic Amplifiers for Context-Independent Expression in the Food-Grade Bacterium Corynebacterium glutamicum

Y ZHAO, Manman Sun, Rodrigo Ledesma‐Amaro, Bin Ye, Alex Xiong Gao, Peng Wang, Farkhod Eshboev
article en

Abstract

is a crucial food-grade (GRAS) bacterial chassis widely utilized for the industrial production of amino acids and nutraceuticals. However, the efficient production of recombinant proteins and secondary metabolites in this host remains limited by context dependence and low translational efficiency. To overcome this, we introduce a 5'-end translationalization strategy. By repurposing passive 5' untranslated regions (5'UTRs) into actively translated fore-cistrons, we converted conventional monocistronic designs into context-independent, leaderless polycistronic designs (PCDs). This assembly of concatenated fore-cistrons functions as a translational amplifier, largely decoupling protein output from mRNA abundance. We validated this platform by optimizing two biomanufacturing paradigms: achieving a 4.07-fold enhanced secretion of OmlA, a porcine vaccine antigen, and boosting biosynthesis of the food-grade pigment indigoidine to 1.20 g/L (a 7.33-fold increase over baselines). Together, this framework establishes a versatile, portable toolkit to overcome translational bottlenecks, enabling robust hyperproduction of recombinant proteins and engineered metabolites in biotechnology.

Journal of Agricultural and Food Chemistry
Chinese Academy of Sciences (CN), Hong Kong University of Science and Technology (HK), Hefei Institutes of Physical Science (CN), High Magnetic Field Laboratory (CN), Artificial Intelligence Research Institute (ES), New Uzbekistan University (UZ), National University of Uzbekistan (UZ), Imperial College London (GB)
National Natural Science Foundation of China, China Postdoctoral Science Foundation, Natural Science Foundation of Anhui Province
Zero hunger
Openalex Percentile: Top 12%
Microbial Metabolic Engineering and Bioproduction
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