Highly active self-sufficient nitration biocatalysts

A fusion protein comprising:

• (i) a TxtE enzyme;
• (ii) an amino acid linker that is 13 amino acids in length;
• and,
• (iii) a catalytic domain of a CYP102A1 (P450BM3) reductase enzyme;
• wherein the linker joins (iii) to a terminus of (i), and wherein the amino acid linker is not set forth in SEQ ID NO:
• 44.

A compound of Formula IX:

• wherein:
• each of Y2 and Y3 is, independently, hydrogen, halogen, substituted or unsubstituted C1-6 alkyl, substituted or unsubstituted C2-6 alkenyl, substituted or unsubstituted C2-6 alkynyl, substituted or unsubstituted, monocyclic, 3-to 6-membered carbocyclyl, substituted or unsubstituted, monocyclic, 3- to 6-membered heterocyclyl, substituted or unsubstituted phenyl, substituted or unsubstituted, monocyclic, 5- to 6-membered heteroaryl, ORAla, N(RAla)2, or SRAla,
• wherein each RAla is independently hydrogen, substituted or unsubstituted acyl, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted carbocyclyl, substituted or unsubstituted heterocyclyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, a nitrogen protecting group when attached to a nitrogen atom, an oxygen protecting group when attached to an oxygen atom, or a sulfur protecting group when attached to a sulfur atom, or two instances of RAla are joined to form a substituted or unsubstituted, heterocyclic ring, or substituted or unsubstituted, heteroaryl ring;
• R1 is H or optionally substituted alkyl;
• R2 is H, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted aryl, or optionally substituted heteroaryl;
• and
• X is NO2;
• or a pharmaceutically acceptable salt thereof.