Metabolic engineering of Escherichia coli to optimize melanin synthesis from glucose.
BACKGROUND
Natural aromatic polymers, mainly melanins, have potential and current applications in the cosmetic, pharmaceutical and chemical industries. The biotechnological production of this class of compounds is based on tyrosinase-dependent conversion of L-tyrosine and other aromatic substrates into melanins. The purpose of this work was to apply metabolic engineering for generating Escherichia coli strains with the capacity to synthesize an aromatic polymer from a simple carbon source.


RESULTS
The strategy was based on the expression in E. coli of the MutmelA gene from Rhizobium etli, encoding an improved mutant tyrosinase. To direct the carbon flow from central metabolism into the common aromatic and the L-tyrosine biosynthetic pathways, feedback inhibition resistant versions of key enzymes were expressed in strains lacking the sugar phosphotransferase system and TyrR repressor. The expressed tyrosinase consumed intracellular L-tyrosine, thus causing growth impairment in the engineered strains. To avoid this issue, a two phase production process was devised, where tyrosinase activity was controlled by the delayed addition of the cofactor Cu. Following this procedure, 3.22 g/L of melanin were produced in 120 h with glucose as carbon source. Analysis of produced melanin by Fourier transform infrared spectroscopy revealed similar characteristics to a pure eumelanin standard.


CONCLUSIONS
This is the first report of a process for producing melanin from a simple carbon source at grams level, having the potential for reducing production cost when compared to technologies employing L-tyrosine as raw material.
María I
MI
Chávez-Béjar
Victor E
VE
Balderas-Hernandez
Aída
A
Gutiérrez-Alejandre
Alfredo
A
Martinez
Francisco
F
Bolívar
Departamento de Ingeniería Celular y Biocatálisis, Instituto de Biotecnología, Universidad Nacional Autónoma de México, Apdo, Postal 510-3, Cuernavaca, Morelos CP 62271, México. gosset@ibt.unam.mx.
Guillermo
G
Gosset
D004926:Escherichia coli / Q000201:enzymology* / Q000235:genetics / Q000378:metabolism; D005947:Glucose / Q000378:metabolism*; D008543:Melanins / Q000096:biosynthesis / Q000138:chemical synthesis*; D060847:Metabolic Engineering; D014442:Monophenol Monooxygenase / Q000378:metabolism*; D010957:Plasmids / Q000378:metabolism; D014443:Tyrosine / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism*
D008543:Melanins; D014443:Tyrosine; D014442:Monophenol Monooxygenase; D005947:Glucose