Simple Synthesis of Blue Dye Indigo Achieved: Engineering Bacillus subtilis P450 Enzyme CYP107J1 into a Hydrogen Peroxide-Driven Form

A research group led by Hideki Kato and Professor Toshiki Furuya at the Tokyo University of Science, in collaboration with Dr. Stephen Bell of the University of Adelaide, has successfully engineered the Bacillus subtilis-derived P450 enzyme CYP107J1 into a hydrogen peroxide-driven enzyme that eliminates the need for electron transport proteins. P450 enzymes are highly valued for their ability to insert oxygen into positions that are difficult to target via conventional organic synthesis, making them promising for pharmaceutical applications. However, their activity typically requires electron donors like NAD(P)H and a series of electron transport proteins, which has historically hindered research. The group introduced two amino acid mutations (Glu251Gln/Thr252Glu) into the active site of CYP107J1, successfully creating a variant that functions without an electron transport system. This engineered enzyme efficiently produces indigo simply by adding indole and hydrogen peroxide. This breakthrough is expected to serve as a catalyst for producing high-value-added substances without the need for complex coenzymes. The findings were published in the international journal 'Microbial Biotechnology' on May 4, 2026.