Abstract
Indigo, one of the most widely used dyes, is mainly produced by chemical processes, which generate amounts of pollutants and need high energy consumption. Microbial production of indigo from indole has attracted much attention; however, the indole oxygenase has never been explored and applied for indigo production. In the present study, the indole oxygenase indAB genes were successfully cloned from Cupriavidus sp. SHE and heterologously expressed in Escherichia coli BL21(DE3) (designated as IND_AB). Strain IND_AB produced primarily indigo in tryptophan medium by high-performance liquid chromatography-mass spectroscopy (HPLC-MS) analysis. The preferable conditions for indigo production were pH 6.5 (normal pH), 30°C, 150rpm, strain inoculation concentration OD600 0.08, and induction with 1mM IPTG at the time of inoculation. The optimal culture medium compositions were further determined as tryptophan 1.0g/L, NaCl 3.55g/L, and yeast extract 5.12g/L based on single-factor experiment and response surface methodology. The highest indigo yield was 307mg/L, which was 4.39-fold higher than the original value. This is the first study investigating indigo production using the indole oxygenase system and the results highlighted its potential in bio-indigo industrial application.
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