Pyomelanin production via heterologous expression of 4-hydroxyphenylpyruvate dioxygenase (HPPD) and construction of HPPD inhibitor screening model

Abstract

Melanin has an increasing market demand in cosmetics, food, medicine as well as aerospace due to its unique properties. Heterologous expression of 4-hydroxyphenylpyruvate dioxygenase (HPPD) from the melanin-producing strain Streptomyces fungicidicus NW-EN1 in Escherichia coli shortened the fermentation cycle of melanin. HPPD catalyzed 4-hydrophenylpyruvate (HPP) to form homologous acid (HGA) and finally form melanin. The purified melanin had the highest absorption peak at 460 nm. Fourier transform infrared spectroscopy and scanning electron microscope scanning showed that the pigment had universal characteristic peaks. The presence of HGA, a predictor of pyomelanin, was identified by high-performance liquid chromatography analysis. The recombinant E. coli produced 804.4 ± 5.9 mg/L pyomelanin within 48 h. Metal ions had a great influence on the production of pyomelanin. Pyomelanin was stable in response to light intensity and had a protective effect against bacteria under UV irradiation. Meanwhile, we utilized the chromogenic effect after whole-cell catalysis to reflect the inhibition of the HPPD inhibitors (mesotrione and isoxaflutole) on HPPD by observing the color change. As a rapid method to test the action of inhibitors, this method is expected to be useful for the development of HPPD-inhibiting herbicides.

Keywords: 4-Hydroxyphenylpyruvate dioxygenase; 4-Hydroxyphenylpyruvate dioxygenase inhibitor; Pyomelanin production; Screening model; UV protection.