Characterization of a flavin-containing monooxygenase from Corynebacterium glutamicum and its application to production of indigo and indirubin
Autor: | Jin-Ho Lee, Sang-Soo Han, Sisi Patricia Lolita Ameria, Hee Sook Kim, Hye Sook Jung, Hak-Sung Kim |
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Rok vydání: | 2015 |
Předmět: |
Indoles
Flavoprotein Gene Expression Bioengineering Flavin-containing monooxygenase Indigo Carmine Applied Microbiology and Biotechnology Indigo Corynebacterium glutamicum chemistry.chemical_compound Indoxyl Enzyme Stability Escherichia coli Cloning Molecular Enzyme Inhibitors Indole test biology Chemistry Spectrum Analysis Temperature General Medicine Hydrogen-Ion Concentration Recombinant Proteins Biochemistry Indigo carmine biology.protein Oxygenases Indirubin Copper Biotechnology |
Zdroj: | Biotechnology letters. 37(8) |
ISSN: | 1573-6776 |
Popis: | 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. 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. The results suggest the cFMO can be used for the microbial production of both indigo and indirubin. |
Databáze: | OpenAIRE |
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