Expanding metabolic pathway for de novo biosynthesis of the chiral pharmaceutical intermediate L-pipecolic acid in Escherichia coli.
Microb Cell Fact
; 16(1): 52, 2017 Mar 27.
Article
en En
| MEDLINE
| ID: mdl-28347340
ABSTRACT
BACKGROUND:
The six-carbon circular non-proteinogenic compound L-pipecolic acid is an important chiral drug intermediate with many applications in the pharmaceutical industry. In the present study, we developed a metabolically engineered strain of Escherichia coli for the overproduction of L-pipecolic acid from glucose.RESULTS:
The metabolic pathway from L-lysine to L-pipecolic acid was constructed initially by introducing lysine cyclodeaminase (LCD). Next, L-lysine metabolic flux from glucose was amplified by the plasmid-based overexpression of dapA, lysC, and lysA under the control of the strong trc promoter to increase the biosynthetic pool of the precursor L-lysine. Additionally, since the catalytic efficiency of the key enzyme LCD is limited by the cofactor NAD+, the intracellular pyridine nucleotide concentration was rebalanced by expressing the pntAB gene encoding the transhydrogenase, which elevated the proportion of LCD with bound NAD+ and enhanced L-pipecolic acid production significantly. Further, optimization of Fe2+ and surfactant in the fermentation process resulted in 5.33 g/L L-pipecolic acid, with a yield of 0.13 g/g of glucose via fed-batch cultivation.CONCLUSIONS:
We expanded the metabolic pathway for the synthesis of the chiral pharmaceutical intermediate L-pipecolic acid in E. coli. Using the engineered E. coli, a fast and efficient fermentative production of L-pipecolic acid was achieved. This strategy could be applied to the biosynthesis of other commercially and industrially important chiral compounds containing piperidine rings.Palabras clave
Texto completo:
1
Banco de datos:
MEDLINE
Asunto principal:
Ácidos Pipecólicos
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Escherichia coli
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Redes y Vías Metabólicas
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Ingeniería Metabólica
Idioma:
En
Revista:
Microb Cell Fact
Asunto de la revista:
BIOTECNOLOGIA
/
MICROBIOLOGIA
Año:
2017
Tipo del documento:
Article