Abstract
Objective
To examine the role of a gene encoding flavin-containing monooxygenase (cFMO) from Corynebacterium glutamicum ATCC13032 when cloned and expressed in Escherichia coli for the production of indigo pigments.
Results
The blue pigments produced by recombinant E. coli were identified as indigo and indirubin. The cFMO was purified as a fused form with maltose-binding protein (MBP). The enzyme was optimal at 25 °C and pH 8. From absorption spectrum analysis, the cFMO was classified as a flavoprotein. FMO activity was strongly inhibited by 1 mM Cu2+ and recovered by adding 1–10 mM EDTA. The enzyme catalyzed the oxidation of TMA, thiourea, and cysteamine, but not glutathione or cysteine. MBP-cFMO had an indole oxygenase activity through oxygenation of indole to indoxyl. The recombinant E. coli produced 685 mg indigo l−1 and 103 mg indirubin l−1 from 2.5 g l-tryptophan l−1.
Conclusion
The results suggest the cFMO can be used for the microbial production of both indigo and indirubin.
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Acknowledgments
This research was supported by Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education, Science and Technology (NRF-2012R1A1A2007229).
Supporting information
Supplementary Table 1—The bacterial strains and plasmids used in this study.
Supplementary Figure 1—Effect of pH (a) and temperature (b) on the NADPH oxidase activity of purified MBO-Cfmo.
Supplementary Figure 2—Absorption spectra of purified MBP-cFMO.
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Ameria, S.P.L., Jung, H.S., Kim, H.S. et al. Characterization of a flavin-containing monooxygenase from Corynebacterium glutamicum and its application to production of indigo and indirubin. Biotechnol Lett 37, 1637–1644 (2015). https://doi.org/10.1007/s10529-015-1824-2
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DOI: https://doi.org/10.1007/s10529-015-1824-2