Bacterial melanin production by heterologous expression of 4‑hydroxyphenylpyruvate dioxygenase from Pseudomonas aeruginosa.

Abstract

Pyomelanin is a reddish-brown pigment produced by bacteria of different genera and plays a variety of physiological roles. Proposals have been regarding the use of pyomelanin in various environmental, industrial and, more recently, cosmetic applications. In Pseudomonas aeruginosa, the enzyme 4‑hydroxyphenylpiruvate dioxygenase (Hpd) converts 4-hydroxyphenylpiruvate into homogentisic acid, which represents the key intermediate for melanin biosynthesis. This work aimed to obtain Escherichia coli cells overexpressing hpd gene from the PAO1 strain to produce large amounts of pyomelanin for biotechnological purposes. The recombinant dioxygenase expression gave E. coli JM109 the ability to produce pyomelanin. A series of biotransformations led us to choose the best experimental conditions for pyomelanin production. Cells were grown at the mid-exponential phase in a mineral medium with added glucose 10 mM as carbon and energy sources and casamino acid 0.2% w/v as an amino acid source. The administration of tyrosine 1 mM after 30 min of exposure to arabinose 1% w/v made it possible to purify 213 mg/L of pyomelanin after 6 days of biotransformation. In addition to the interesting biotechnological outcomes, the resulting expression system supports the correlation between the hpd gene from P. aeruginosa PAO1 and pyomelanin synthesis.