Construction of a genome-engineered stable 5-aminolevulinic acid producing Corynebacterium glutamicum by increasing succinyl-CoA supply
Year: 2025
Type: article
Abstract: 5-Aminolevulinic acid (5-ALA), a versatile precursor for tetrapyrrole derivatives (heme/chlorophyll/cobalamin), drives advancing microbial cell factories to meet growing biomedical and industrial demands. However, there remains two challenges limits yield and scalability: conventional plasmid-dependent systems suffer from genetic instability, antibiotic reliance and metabolic burden, and the lack of fine regulation of succinyl-CoA. In this study, multiple copies of hemAC132A of the heterologous C4 pathway were firstly integrated on the genome. For fine regulating the supply of succinyl-CoA, the genes related to the tricarboxylic acid cycle (TCA cycle) oxidation branch pathway were combinatorially screened. The optimal combination of icd and lpd was confirmed by ribosome binding site (RBS) engineering, which was integrated on the genome with optimized expression intensity. Succinyl-CoA supply was further increased by genome integration and expression optimization of key CoA biosynthetic gene coaA, pantothenic acid synthesis-related gene panB-panC, and β-alanine synthesis-related gene panD. The optimized genomically stable chasis achieved a high 5-ALA production of 6.39 ± 0.20 g/L, which was 8.63-fold higher than the single hemAC132A copy strain A1 (0.74 ± 0.07 g/L). From these findings, a stable and high-yield 5-ALA synthetic strain was successfully constructed, providing a new strategy for production of biochemicals derived from succinyl-CoA in C. glutamicum.
Institutions: Tianjin University, Ningxia University
Language: English
Subfield: Molecular Biology
Domain: Life Sciences
SDG: Zero hunger
Open Access status: gold
Estimated APC paid: $1,500
Awards: G474876263, G6649598041
OpenAlex ID: W4410908209