Abstract
Objective
To construct a strain of Corynebacterium glutamicum capable of efficiently producing 5-aminolevulinic acid (5-ALA) via the C4 pathway by modification of serine and glycine pathway using glucose as sole carbon source.
Results
The recombinant C. glutamicum strain AP2 harboring a codon-optimized hemA gene from Rhodobacter sphaeroides was used as host strain for 5-ALA production. A plasmid harboring the serine operon, which contained serB, serC and the site-specific mutant serA Δ197, was constructed and introduced into C. glutamicumAP2, leading to an increase of 70% in 5-ALA production. Further overexpression of the glyA gene increased production of 5-ALA by 150% over the control. 5-ALA production was thus significantly enhanced by engineering the glycine biosynthetic pathway. C.glutamicum AG3 produced 3.4 ± 0.2 g 5-ALA/l in shake-flask cultures in CGIIIM medium with the addition of 7.5 g glycine/l.
Conclusion
This is the first report of remodeling the serine and glycine biosynthetic pathway to improve the production of 5-ALA in C. glutamicum.
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Acknowledgements
This work was supported by the Natural Science Foundation of Tianjin (No. 15JCQNJC06000).
Supporting information
Supplementary Table 1—Strains and plasmids used.
Supplementary Table 2—Primers used.
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Zou, Y., Chen, T., Feng, L. et al. Enhancement of 5-aminolevulinic acid production by metabolic engineering of the glycine biosynthesis pathway in Corynebacterium glutamicum . Biotechnol Lett 39, 1369–1374 (2017). https://doi.org/10.1007/s10529-017-2362-x
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DOI: https://doi.org/10.1007/s10529-017-2362-x