Construction of metabolic operons catalyzing the de novo biosynthesis of indigo in Escherichia coli.
The efficient production of the textile dye indigo by fermentation has been a goal since the early 1980's when the first bacterial strains capable of this synthesis were constructed. We report here the development of a recombinant microorganism that directly synthesizes indigo from glucose. This construction involved the cloning and genetic manipulation of at least 9 genes and modifications of the fermentation medium to help stabilize the biosynthetic activity. Directed genetic changes in two operons caused significant increases in reaction rates and in the stability of the catalytic enzymes. This example of whole cell catalysis by a recombinant Escherichia coli represents a novel and environmentally sound approach to the synthesis of a high value specialty chemical.
Murdock
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Amgen, Inc., Thousand Oaks, CA 93120.
Ensley
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Serdar
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Thalen
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D001483:Base Sequence; D003001:Cloning, Molecular; D049308:Dioxygenases; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D005285:Fermentation; D005288:Ferredoxins / Q000235:genetics; D015870:Gene Expression; D005947:Glucose / Q000378:metabolism; D007203:Indigo Carmine; D007211:Indoles; D008969:Molecular Sequence Data; D009097:Multienzyme Complexes / Q000235:genetics / Q000378:metabolism; D016297:Mutagenesis, Site-Directed; D009876:Operon; D010105:Oxygenases / Q000235:genetics / Q000378:metabolism; D010957:Plasmids; D016958:Pseudomonas putida / Q000201:enzymology; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000235:genetics
D005288:Ferredoxins; D007211:Indoles; D009097:Multienzyme Complexes; D011994:Recombinant Proteins; D007203:Indigo Carmine; D010105:Oxygenases; D049308:Dioxygenases; C038703:naphthalene dioxygenase; D005947:Glucose