POTENTIAL USE OF BACTERIAL FLAVING-CONTAINING MONOOXYGENASES FOR INDIGO AND INDIRUBIN PRODUCTION IN ESCHERICHIA COLI

Salta

Congreso; Joint LV Annual SAIB Meeting and XIV PABMB Congress; 2019

SAIB

Indole alkaloids are a large group of secondary metabolites. Indigoid compounds such as indigo and indirubin have been employed as natural dyes and drugs since ancient times. Indigo is a commercially important blue pigment that is used to dye both cotton and wool, while indirubin is currently used as a drug to treat chronic granulocytic leukemia and is a potential drug against cancer and Alzheimer?s disease as well. In this work we analyzed the potential use of flaving-containing monooxygenases (FMO) from Roseovarius sp. and/or Methylocella silvestris for indigo and indirubin production in Escherichia coli. The FMO coding sequences were E. coli codon optimized and synthesized having a C-terminal his-tag coding sequence. Overexpression of FMOs in E. coli produced a large amount of the recombinant proteins. The analysis of the purified proteins showed that his-tagged FMOs are fully active. However, production of indigo and indirubin was only observed under specific culture conditions. Optimization of the upstream conditions (culture media composition, proteins overexpression time, addition of tryptophan, temperature of growth, etc) allowed the production of a large amount of indigo and indirubin in E. coli. The potential use of these enzymes for the commercial production of indigo, indirubin and other indole derivatives of biotechnological interest will be discussed.