Bio-indigo Production using Wild-type Acinetobacter sp. and Indole-3-acetate Monooxygenase (iacA) Expressed in Escherichia coli

This study reports the identification of a novel bio-indigo-producing bacterium from soil sources. Blue-colored colonies were first screened and isolated from a screening plate containing M9 minimal medium and 1.0 mM indole. The blue colony was selected among various colonies and identified as an Ac...

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Bibliographic Details
Published in:Biotechnology and bioprocess engineering 2023, 28(2), , pp.281-288
Main Authors: Ahn, Sooyeon, Park, SeoA, Kumar, Pradeep, Choi, Kwon-Young
Format: Article
Language:English
Subjects:
Acetic acid
Acinetobacter
Biotechnology
Chemistry
Chemistry and Materials Science
Chromatography
Colonies
Dyes
E coli
Escherichia coli
Indigo
Indoles
Industrial and Production Engineering
Ions
Liquid chromatography
Mass spectrometry
Mass spectroscopy
Research Paper
Sequence analysis
Soil bacteria
Soil microorganisms
Substrates
Thin layer chromatography
생물공학
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Summary:This study reports the identification of a novel bio-indigo-producing bacterium from soil sources. Blue-colored colonies were first screened and isolated from a screening plate containing M9 minimal medium and 1.0 mM indole. The blue colony was selected among various colonies and identified as an Acinetobacter species. The purified blue dye exhibited a distinct spectral feature of λ max at 490 nm. The structure of the dye was then analyzed. Thin-layer chromatography separation and liquid chromatography/mass spectrometry analysis confirmed that the blue dye was indigo (with R f and m/z values of 0.8 and 263.4, respectively). Wild-type Acinetobacter sp. could produce bio-indigo up to 1.018 ± 0.013 mg/L in an M9 minimal medium supplemented with 1.0 mM indole. Next, the genes involved in the production of indigo were investigated using sequence analysis and by comparing them with those in related Acinetobacter species. The indole-3-acetate monooxygenase-encoding gene iacA was found to be responsible for indigo synthesis from indole. The iacA gene was then amplified and expressed in Escherichia coli BL21(DE3), and the recombinant E. coli strain could produce bio-indigo at levels up to 0.291 ± 0.027 g/L over 24 h. Indigo production was highly dependent on indole substrate feeding. These findings may facilitate the industrial bioprocess of bio-indigo production.
ISSN:1226-8372
1976-3816
DOI:10.1007/s12257-022-0163-0