TY - JOUR
T1 - Accelerated 5-aminolevulinic acid biosynthesis by coupling aconitase and ALA synthase in engineered Escherichia coli
AU - Lai, Ping Hao
AU - Ng, I. S.
N1 - Publisher Copyright:
© 2024 Elsevier B.V.
PY - 2024/9
Y1 - 2024/9
N2 - In recent years, 5-aminolevulinic acid (5-ALA) has attracted significant interest due to its roles as a photodynamic prodrug and an antiviral agent. In this study, we present a new approach using aconitase A from Escherichia coli Nissle 1917 (EcNAcnA), renowned for its exceptional activity and conjunction with ALA synthase from Rhodobacter capsulatus (RcALAS) to enhance 5-ALA production in an engineered chassis. Expression of EcNAcnA and RcALAS via dual plasmids led to a 59 % increase in 5-ALA yield, reaching up to 6.645 g/L. Diverse 5-ALA production levels were observed with different combinations of promoters and replication origins for both genes. Subsequently, an all-in-one plasmid with a high copy number, designated as RcNN, was introduced into the genomic engineering RcI strain. This resulted in the production of 24.5 g/L 5-ALA with a productivity of 0.907 g/L/h in a bioreactor under pH control and glucose feeding over 27 h. To the best of our knowledge, this is the first study to enhance 5-ALA biosynthesis by applying a superior aconitase variant from E. coli Nissle 1917, which enhances isocitrate production in the tricarboxylic acid (TCA) cycle and alleviates reactive oxygen species (ROS), thereby promoting 5-ALA accumulation effectively.
AB - In recent years, 5-aminolevulinic acid (5-ALA) has attracted significant interest due to its roles as a photodynamic prodrug and an antiviral agent. In this study, we present a new approach using aconitase A from Escherichia coli Nissle 1917 (EcNAcnA), renowned for its exceptional activity and conjunction with ALA synthase from Rhodobacter capsulatus (RcALAS) to enhance 5-ALA production in an engineered chassis. Expression of EcNAcnA and RcALAS via dual plasmids led to a 59 % increase in 5-ALA yield, reaching up to 6.645 g/L. Diverse 5-ALA production levels were observed with different combinations of promoters and replication origins for both genes. Subsequently, an all-in-one plasmid with a high copy number, designated as RcNN, was introduced into the genomic engineering RcI strain. This resulted in the production of 24.5 g/L 5-ALA with a productivity of 0.907 g/L/h in a bioreactor under pH control and glucose feeding over 27 h. To the best of our knowledge, this is the first study to enhance 5-ALA biosynthesis by applying a superior aconitase variant from E. coli Nissle 1917, which enhances isocitrate production in the tricarboxylic acid (TCA) cycle and alleviates reactive oxygen species (ROS), thereby promoting 5-ALA accumulation effectively.
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U2 - 10.1016/j.bej.2024.109419
DO - 10.1016/j.bej.2024.109419
M3 - Article
AN - SCOPUS:85197255289
SN - 1369-703X
VL - 209
JO - Biochemical Engineering Journal
JF - Biochemical Engineering Journal
M1 - 109419
ER -