Production of 5-aminolevulinic acid from glutamate by overexpressing HemA1 and pgr7 from Arabidopsis thaliana in Escherichia coli
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The important metabolic intermediate 5-aminolevulinic acid (ALA) is useful for cancer treatment or plant growth regulation and has consequently received much attention. In this study, we introduced the HemA1 and pgr7 genes from the higher plant Arabidopsis thaliana into recombinant Escherichia coli to overproduce extracellular 5-aminolevulinic acid via the C5 pathway. In the E. coli BL21 (DE3) strain background, the ALA concentration of the strain expressing both HemA1 and pgr7 was the highest and reached 3080.62 mg/L. Among the 7 tested hosts, ALA production was the highest in E. coli Transetta (DE3). In E. coli Transetta GTR/GBP, the expression levels of zwf, gnd, pgl and RhtA were upregulated. Glutamate induced the expression of the GltJ, GltK, GltL and GltS genes that are in involved in glutamate uptake. The recombinant E. coli Transetta GTR/GBP was able to produce 7642 mg/L ALA in modified minimal medium supplemented with 10 g/L glutamate and 15 g/L glucose after 48 h of fermentation at 22 °C. The results provide persuading evidence for the efficient production of ALA from glucose and glutamate in E. coli expressing A. thaliana HemA1 and pgr7. Further optimization of the fermentation process should be done to improve the ALA production to an industrially relevant level.
Keywords5-Aminolevulinic acid Arabidopsis thaliana Escherichia coli Glutamyl-tRNAGlu reductase
This work was financially supported by a grant from the National Natural Science Foundation of China (31500243), and a grant of the national basic research program of China (Z109021566).
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Conflict of interest
The authors declare that they have no conflict of interest.
Research involving human and animal participants
This manuscript does not contain any studies with human participants or animals performed by any of the authors.
- Mikolajewska P, Donnelly RF, Garland MJ et al (2010) Microneedle pre-treatment of human skin improves 5-aminolevulininc acid (ALA)- and 5-aminolevulinic acid methyl ester (MAL)-induced PpIX production for topical photodynamic therapy without increase in pain or erythema. Pharm Res 27:2213–2220. https://doi.org/10.1007/s11095-010-0227-2 CrossRefPubMedGoogle Scholar
- Mills-Davies N, Butler D, Norton E et al (2017) Structural studies of substrate and product complexes of 5-aminolaevulinic acid dehydratase from humans, Escherichia coli and the hyperthermophile Pyrobaculum calidifontis. Acta Crystallogr D 73:9–21. https://doi.org/10.1107/S2059798316019525 CrossRefGoogle Scholar
- Saikeur A, Choorit W, Prasertsan P, Kantachote D, Sasaki K (2009) Influence of precursors and inhibitor on the production of extracellular 5-aminolevulinic acid and biomass by Rhodopseudomonas palustris KG31. Biosci Biotechnol Biochem 73:987–992. https://doi.org/10.1271/bbb.80682 CrossRefPubMedGoogle Scholar
- Sundara Sekar B, Seol E, Park S (2017) Co-production of hydrogen and ethanol from glucose in Escherichia coli by activation of pentose-phosphate pathway through deletion of phosphoglucose isomerase (pgi) and overexpression of glucose-6-phosphate dehydrogenase (zwf) and 6-phosphogluconate dehydrogenase (gnd). Biotechnol Biofuels 10:85. https://doi.org/10.1186/s13068-017-0768-2 CrossRefPubMedPubMedCentralGoogle Scholar