Optimum melanin production using recombinant Escherichia coli.
OBJECTIVE
A parametric study was conducted to define optimum conditions to achieve high yields in the conversion of tyrosine to eumelanin (EuMel) using recombinant Escherichia coli.


RESULTS
Escherichia coli W3110 (pTrcMutmelA) expressing the tyrosinase coding gene from Rhizobium etli and glucose-mineral media were used to transform tyrosine into EuMel. Batch aerobic fermentor cultures were performed to study the effect of temperature, pH and inducer concentration (isopropyl-D-thio-galactopyranoside) on melanin production. Under optimum conditions, 0.1 mmol l(-1) of isopropyl-D-thio-galactopyranoside, temperature of 30 degrees C, and changing pH from 7.0 to 7.5 during the production phase, a 100% conversion of tyrosine into EuMel is obtained. Furthermore, tyrosine feeding allowed us to obtain the highest level (6 g l(-1)) of EuMel produced by recombinant E. coli reported until now.


CONCLUSIONS
The most important factors affecting melanin formation and hence influencing the rate and efficiency in the conversion of tyrosine into EuMel in this system, are the temperature and pH.


CONCLUSIONS
Maximum theoretical yield was obtained using a simple culture process and mineral media to convert tyrosine (a medium value compound) into melanin, a high value compound. The process reported here avoids the use of purified tyrosinase, expensive chemical methods or the cumbersome extraction of this polymer from animal or plant tissues.
Lagunas-Muñoz
V H
VH
Departamento de Ingeniería Celular y Biocatálisis, Instituto de Biotecnología, Universidad Nacional Autónoma de México, Cuernavaca, Morelos, Mexico.
Cabrera-Valladares
N
N
Bolívar
F
F
Gosset
G
G
Martínez
A
A
D000667:Ampicillin / Q000494:pharmacology; D000668:Ampicillin Resistance; D000900:Anti-Bacterial Agents / Q000494:pharmacology; D019149:Bioreactors; D003470:Culture Media; D004305:Dose-Response Relationship, Drug; D004926:Escherichia coli / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*; D005947:Glucose; D006863:Hydrogen-Ion Concentration; D007544:Isopropyl Thiogalactoside / Q000494:pharmacology; D008543:Melanins / Q000096:biosynthesis*; D015202:Protein Engineering / Q000379:methods; D011994:Recombinant Proteins / Q000096:biosynthesis*; D042163:Rhizobium etli / Q000235:genetics; D013696:Temperature; D014443:Tyrosine / Q000235:genetics / Q000378:metabolism
D000900:Anti-Bacterial Agents; D003470:Culture Media; D008543:Melanins; D011994:Recombinant Proteins; C041877:eumelanin; D007544:Isopropyl Thiogalactoside; D014443:Tyrosine; D000667:Ampicillin; D005947:Glucose