Biotransformation of indole by whole cells of recombinant biphenyl dioxygenase and biphenyl-2,3-dihydrodiol-2,3-dehydrogenase

The introduction of hydroxyl groups into indole molecule by different mono- and dioxygenases leads to the production of indigo. As a well-known biocatalyst, biphenyl dioxygenase possessed the ability to transform indole to indigo. However, there has been little information about this enzymatic transformation process. In this study, the genes encoding biphenyl dioxygenase (BphA) and biphenyl-2,3-dihydrodiol 2,3-dehydrogenase (BphB) were cloned from Dyella ginsengisoli LA-4 and heterologously expressed in Escherichia coli (DE3) (designated as AB No). The feasibility of indole transformation to indigo by strain BphA(_LA-4) was predicted by molecular docking studies. The biotransformation ratios of indole (100 mg/L) reached the maximum (95%) when cells were induced at 15 degrees C with 0.25 mM IPTG in M9 medium. In addition, 44 mg/L indigo was produced from 200 mg/L indole when supplied with 0.28 g/L of biomass and 0.2% (w/v) glucose. HPLC-MS was used to identify the products, which showed that indigo was the major product. Meanwhile, indirubin and isatin were also identified during the transformation process. Furthermore, the pathway of indole transformation by strain AB(_IND) was also proposed. (C) 2013 Elsevier B.V. All rights reserved.