Stable and Efficient Biosynthesis of 5-Aminolevulinic Acid Using Plasmid-Free Escherichia coli

ABSTRACT We found that Escherichia coli tolC mutants showed increased sensitivity to 5-aminolevulinic acid (ALA), a precursor of porphyrins. The tolC mutant cells grown in the presence of ALA showed a reddish brown color under visible light and a strong red fluorescence under near-UV irradiation. Fluorescence spectrometry and high-performance liquid chromatography analysis showed that the tolC mutant cells grown in the presence of ALA accumulated a large amount of coproporphyrin(ogen) intracellularly. In contrast, the wild-type cells produced coproporphyrin extracellularly. The tolC mutant cells grown in the presence of ALA, which were capable of surviving in the dark, were killed by near-UV irradiation, suggesting that the intracellular coproporphyrin(ogen) renders these cells photosensitive. These results suggest that the TolC-dependent efflux system is involved in the exclusion of porphyrin(ogen)s in E. coli.

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Cloning and characterization of genes involved in the biosynthesis of δ-aminolevulinic acid in Escherichia coli

Gene ◽

10.1016/0378-1119(92)90170-t ◽

1992 ◽

Vol 121 (1) ◽

pp. 127-132 ◽

Cited By ~ 10

Author(s):

Ikemi Masahisa ◽

Murakami Katsuji ◽

Hashimoto Masamichi ◽

Yoshikatsu Murooka

Keyword(s):

Escherichia Coli ◽

Aminolevulinic Acid

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Production of uroporphyrinogen III, which is the common precursor of all tetrapyrrole cofactors, from 5-aminolevulinic acid by Escherichia coli expressing thermostable enzymes

Applied Microbiology and Biotechnology ◽

10.1007/s00253-013-4904-z ◽

2013 ◽

Vol 97 (16) ◽

pp. 7337-7344 ◽

Cited By ~ 3

Author(s):

Aiko Hibino ◽

René Petri ◽

Jochen Büchs ◽

Hisao Ohtake

Keyword(s):

Escherichia Coli ◽

Aminolevulinic Acid ◽

Thermostable Enzymes ◽

Common Precursor ◽

The Common

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Occurrence of Two 5-Aminolevulinate Biosynthetic Pathways in Streptomyces nodosus subsp. asukaensis Is Linked with the Production of Asukamycin

Journal of Bacteriology ◽

10.1128/jb.01919-05 ◽

2006 ◽

Vol 188 (14) ◽

pp. 5113-5123 ◽

Cited By ~ 19

Author(s):

Miroslav Petříček ◽

Kateřina Petříčková ◽

Libor Havlíček ◽

Jürgen Felsberg

Keyword(s):

Escherichia Coli ◽

Enzyme Activity ◽

Aminolevulinic Acid ◽

Streptomyces Coelicolor ◽

Biosynthetic Pathways ◽

Tetrapyrrole Biosynthesis ◽

Streptomyces Nodosus ◽

Encoding Gene

ABSTRACT We report the results of cloning genes for two key biosynthetic enzymes of different 5-aminolevulinic acid (ALA) biosynthetic routes from Streptomyces. The genes encode the glutamyl-tRNAGlu reductase (GluTR) of the C5 pathway and the ALA synthase (ALAS) of the Shemin pathway. While Streptomyces coelicolor A3(2) synthesizes ALA via the C5 route, both pathways are operational in Streptomyces nodosus subsp. asukaensis, a producer of asukamycin. In this strain, the C5 route produces ALA for tetrapyrrole biosynthesis; the ALA formed by the Shemin pathway serves as a precursor of the 2-amino-3-hydroxycyclopent-2-enone moiety (C5N unit), an antibiotic component. The growth of S. nodosus and S. coelicolor strains deficient in the GluTR genes (gtr) is strictly dependent on ALA or heme supplementation, whereas the defect in the ALAS-encoding gene (hemA-asuA) abolishes the asukamycin production in S. nodosus. The recombinant hemA-asuA gene was expressed in Escherichia coli and in Streptomyces, and the encoded enzyme activity was demonstrated both in vivo and in vitro. The hemA-asuA gene is situated within a putative cluster of asukamycin biosynthetic genes. This is the first report about the cloning of genes for two different ALA biosynthetic routes from a single bacterium.

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