Unveiling the Effect of <i>NCgl0580</i> Gene Deletion on 5-Aminolevulinic Acid Biosynthesis in <i>Corynebacterium glutamicum</i>

5-Aminolevulinic acid (5-ALA) has recently received much attention for its wide applications in medicine and agriculture. In this study, we investigated the effect of <i>NCgl0580</i> in <i>Corynebacterium glutamicum</i> on 5-ALA biosynthesis as well as its possible mechanism. It was found that the overexpression of <i>NCgl0580</i> increased 5-ALA production by approximately 53.3%. Interestingly, the knockout of this gene led to an even more significant 2.49-fold increase in 5-ALA production. According to transcriptome analysis and functional validation of phenotype-related targets, the deletion of <i>NCgl0580</i> brought about considerable changes in the transcript levels of genes involved in central carbon metabolism, leading to fluxes redistribution toward the 5-ALA precursor succinyl-CoA as well as ATP-binding cassette (ABC) transporters affecting 5-ALA biosynthesis. In particular, the positive effects of enhanced sugar transport (by overexpressing <i>NCgl1445</i> and <i>iolT1</i>), glycolysis (by overexpressing <i>pyk2</i>), iron uptake (by overexpressing <i>afuABC</i>), and phosphate uptake (by overexpressing <i>pstSCAB</i> and <i>ugpQ</i>) on 5-ALA biosynthesis were demonstrated for the first time. Thus, the transcriptional mechanism underlying the effect of <i>NCgl0580</i> deletion on 5-ALA biosynthesis was elucidated, providing new strategies to regulate the metabolic network of <i>C. glutamicum</i> to achieve a further increase in 5-ALA production.