title: Characterization of a flavin-containing monooxygenase from Corynebacterium glutamicum and its application to production of indigo and indirubin.
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 Cu(2+) 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).
  
  
  CONCLUSIONS
  The results suggest the cFMO can be used for the microbial production of both indigo and indirubin.
authors: Food Technology Department, Faculty of Food Science & Technology, Universitas Pelita Harapan, 1100, Jl.M.H. Thamrin Boulevard Raya, Tangerang, 15811, Indonesia.
authors: Sisi Patricia Lolita
authors: SP
authors: Ameria
authors: Hye Sook
authors: HS
authors: Jung
authors: Hee Sook
authors: HS
authors: Kim
authors: Sang Soo
authors: SS
authors: Han
authors: Hak Sung
authors: HS
authors: Kim
authors: Jin Ho
authors: JH
authors: Lee
mesh_terms: D003001:Cloning, Molecular; D003300:Copper / Q000378:metabolism; D048230:Corynebacterium glutamicum / Q000201:enzymology* / Q000235:genetics; D004791:Enzyme Inhibitors / Q000378:metabolism; D004795:Enzyme Stability; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D006863:Hydrogen-Ion Concentration; D007203:Indigo Carmine / Q000378:metabolism*; D007211:Indoles / Q000378:metabolism; D010105:Oxygenases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D013057:Spectrum Analysis; D013696:Temperature
chemical_list: D004791:Enzyme Inhibitors; D007211:Indoles; D011994:Recombinant Proteins; D003300:Copper; D007203:Indigo Carmine; D010105:Oxygenases; C020066:dimethylaniline monooxygenase (N-oxide forming); C027185:indirubin