Enhancement of naphthalene dioxygenase activity during microbial indigo production

A process for enhancing naphthalene dioxygenase activity in a microorganism comprising the step of:

• growing microorganism in a culture broth containing an amount of iron effective to enhance said naphthalene dioxygenase activity to produce indigo from indole, wherein said organism has DNA sequences that express a naphthalene dioxygenase enzyme which comprises reductase, ferredoxin and iron-sulfur polypeptides.

A process for enhancing naphthalene dioxygenase activity to produce indigo from indole in an organism comprising the step of:

• transforming a microorganism with a DNA sequence encoding a naphthalene dioxygenase ferredoxin analog having Cys at position 47, Ile at position 52 and Arg at position 67 in said naphthalene dioxygenase ferredoxin analog amino acid sequence, wherein said microorganism has DNA sequences that express a naphthalene dioxygenase enzyme which comprises reductase, ferredoxin and iron-sulfur polypeptides.

A DNA sequence encoding a naphthalene dioxygenase ferredoxin analog having Cys at position 47, Ile at position 52 and Arg at position 67 in said naphthalene dioxygenase ferredoxin analog amino acid sequence.

A plasmid comprising:

• a DNA sequence encoding a naphthalene dioxygenase ferredoxin analog having Cys at position 47, Ile at position 52 and Arg at position 67 in said naphthalene dioxygenase ferredoxin analog amino acid sequence.

A host cell comprising a DNA sequence encoding a naphthalene dioxygenase ferredoxin analog having Cys at position 47, Ile at position 52 and Arg at position 67 in said naphthalene dioxygenase ferredoxin analog amino acid sequence.

A plasmid comprising:

• a DNA sequence encoding at least two naphthalene dioxygenase ferredoxin polypeptides for each naphthalene dioxygenase reductase polypeptide and iron-sulfur polypeptide.

A host cell comprising:

• a DNA sequence encoding at least two naphthalene dioxygenase ferredoxin polypeptides for each naphthalene dioxygenase reductase polypeptide and iron-sulfur polypeptide.

A process for enhancing naphthalene dioxygenase activity to microbially produce indigo from indole comprising:

• growing a microorganism in a culture broth containing an amount of iron effective to enhance said naphthalene dioxygenase activity to produce indigo from indole, wherein said microorganism is transformed with Pseudomonas putida DNA sequences encoding a naphthalene dioxygenase enzyme.

A process for enhancing naphthalene dioxygenase activity to microbially produce indigo from indole comprising:

• transforming a microorganism with a modified Pseudomonas putida DNA sequence encoding at least two naphthalene dioxygenase ferredoxin polypeptides for each naphthalene dioxygenase reductase polypeptide and iron-sulfur polypeptide.

A process for enhancing naphthalene dioxygenase activity to microbially produce indigo from indole comprising:

• transforming a microorganism with a modified Pseudomonas putida DNA sequence encoding naphthalene dioxygenase ferredoxin analog having Cys at position 47, Ile at position 52 and Arg at position 67 in said naphthalene dioxygenase ferredoxin analog amino acid sequence, reductase and iron-sulfur polypeptides.