Patent ID: 11879144
Assignee: UNIVERSITY OF FLORIDA RESEARCH FOUNDATION, INC.
Field: Biotechnology (Chemistry)
Classification: CPC C | IPC C

Claim 0:
1. A method for producing a nitrated L-tryptophan or nitrated L-tryptophan analogue, the method comprising:
(i) introducing into a bacterial cell culture comprising a recombinant bacterial cell comprising one or more isolated nucleic acids engineered to express:
a fusion protein comprising a TxtE enzyme linked to a catalytic domain of a CYP102A1 (P450BM3) reductase enzyme via an amino acid linker sequence that is between 14 and 27 amino acids in length, wherein the fusion protein is a TB14 fusion protein having the sequence set forth in SEQ ID NO: 1;
a nitric oxide synthase (NOS) enzyme; and, optionally
a glucose dehydrogenase (GDH) enzyme, one or more L-Trp molecules and/or one or more L-Trp analogue molecules;

(ii) growing the bacterial cell culture under conditions under which the fusion protein catalyzes a nitration reaction which produces one or more nitrated L-Trp molecules and/or one or more nitrated L-Trp analog molecules; and, optionally,
(iii) isolating nitrated L-Trp molecules and/or nitrated L-Trp analog molecules.