Alkane oxidation by modified hydroxylases

An isolated or recombinant polypeptide comprising residues 456-1048 of the amino acid sequence set forth in SEQ ID NO:

• 1 with up to 65 conservative amino acid substitutions excluding residues 464, 631, 645, 710 and 968, wherein the amino acid sequence comprises residues selected from the group consisting of:
• (a) at position 464, a Z1, Z3, Z4, or Z5 amino acid residue;
• (b) at positions 631 and 710, a Z1 amino acid residue;
• (c) at position 645, a Z3 amino acid residue;
• and
• (d) at position 968, a Z2 amino acid residue,
• wherein:
• Z1 is an amino acid residue selected from the group consisting of glycine (G), asparagine (N), glutamine (Q), serine(S), threonine (T), tyrosine (Y), and cysteine (C);
• Z2 is an amino acid residue selected from the group consisting of alanine (A), valine (V), leucine (L), isoleucine (I), proline (P), and methionine (M);
• Z3 is an amino acid residue selected from the group consisting of lysine (K), and arginine (R);
• Z4 is an amino acid residue selected from the group consisting of tyrosine (Y), phenylalanine (F), tryptophan (W), and histidine (H);
• and is an amino acid residue selected from the group consisting of threonine (T), valine (V), and isoleucine (I), wherein when the isolated or recombinant polypeptide is operably linked to the heme domain of a cytochrome P450 enzyme to form a chimeric polypeptide the chimeric polypeptide has cytochrome P450 activity.