Melanin production in Escherichia coli from a cloned tyrosinase gene.
We have cloned and functionally expressed a tyrosinase gene from Streptomyces antibioticus in Escherichia coli under the control of an inducible bacteriophage T7 promoter. Recombinant E. coli cells containing the induced tyrosinase gene produced melanin pigments on agar plates and in liquid culture when supplemented with copper and tyrosine. The expression of an additional open reading frame from the mel gene locus of S. antibioticus was required for high-level melanin production in E. coli. Our results also show that it is possible to screen other classes of precursor compounds for incorporation into melanin pigments with unique colors and other biochemical features. In addition, it may be possible to screen for enhanced melanin production in the absence of added precursors to identify overproducing mutants in the amino acid biosynthetic pathways of E. coli. The ability to screen for a melanin phenotype in recombinant E. coli provides new opportunities for production of novel melanins and for protein engineering of tyrosinases with altered catalytic properties.
della-Cioppa
G
G
Biosource Genetics Corporation, Vacaville, CA 95688.
Garger
S J
SJ
Sverlow
G G
GG
Turpen
T H
TH
Grill
L K
LK
D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism; D008543:Melanins / Q000096:biosynthesis* / Q000235:genetics; D014442:Monophenol Monooxygenase / Q000235:genetics* / Q000378:metabolism; D016366:Open Reading Frames; D010641:Phenotype; D010957:Plasmids; D013303:Streptomyces antibioticus / Q000201:enzymology / Q000235:genetics; D014169:Transformation, Bacterial
D008543:Melanins; D014442:Monophenol Monooxygenase