Synthetic indigo is among the most important industrial dyes. Unfortunately, synthetic indigo has raised environmental concern due to the use of pollutant and hazardous chemicals. In response, green chemistry aims to find new environmentally friendly and economically attractive synthesis methods. Accordingly, this work describes the enzymatic synthesis of indigo and its derivatives starting from indole. A variant of the heme domain of cytochrome P450 from Bacillus megaterium, CYP_BM3F87A, was used to catalyze the synthesis of indigo and three di-substituted derivatives. Substrates transformed by a peroxygenase reaction were indole, 4-bromoindole, 5-methoxyindole and 7-methoxyindole. The kinetic data fitted to Hill's mathematical model with k_cat values of 3.51, 0.94, 4.72, and 4.28 min⁻¹ for each substrate, respectively. In all cases, colored products were obtained that were characterized by spectrometric techniques. Furthermore, the CYP_BM3F87A enzyme was immobilized on magnetic nanoparticles that exhibited catalytic rate in the synthesis of indigo from indole (k_cat 0.72 min⁻¹). Our results show that enzymatic synthesis of industrial dyes without the use of expensive cofactors offers an attractive and plausible alternative to conventional chemical synthesis with a lower environment impact.

合成靛蓝是最重要的工业染料之一。不幸的是，由于使用了污染物和有害化学物质，合成靛蓝引起了人们对环境的关注。作为回应，绿色化学旨在寻找新的对环境友好且经济上有吸引力的合成方法。因此，这项工作描述了从吲哚开始的靛蓝及其衍生物的酶促合成。来自巨大芽孢杆菌的细胞色素P450血红素结构域的变体CYP _BM3 F87A被用于催化靛蓝和三种二取代衍生物的合成。通过过加氧酶反应转化的底物是吲哚，4-溴吲哚，5-甲氧基吲哚和7-甲氧基吲哚。用k _cat将动力学数据拟合到Hill的数学模型每个底物的值分别为3.51、0.94、4.72和4.28 min ^-1。在所有情况下，均获得了用光谱技术表征的有色产品。此外，将CYP _BM3 F87A酶固定在磁性纳米颗粒上，该磁性纳米颗粒在从吲哚合成靛蓝中表现出催化速率（k _cat 0.72 min ^-1）。我们的结果表明，不使用昂贵的辅因子的酶法合成工业染料提供了一种对常规化学合成的有吸引力且可行的替代方法，且对环境的影响较小。