28-01-2016 дата публикации
Номер: US20160024482A1
Принадлежит:
This invention relates to modified hydroxylases. The invention further relates to cells expressing such modified hydroxylases and methods of producing hydroxylated alkanes by contacting a suitable substrate with such cells. 1. An isolated polypeptide comprising the amino acid sequence set forth in SEQ ID NO:1 with up to 50 conservative amino acid substitutions excluding residues 47 , 52 , 74 , 78 , 82 , 94 , 142 , 175 , 184 , 188 , 205 , 226 , 236 , 252 , 255 , 290 , 328 , 353 , 366 , 464 and 710 , wherein if the residue selected from 47 , 52 , 74 , 78 , 94 , 142 , 175 , 184 , 188 , 205 , 226 , 236 , 252 , 255 , 290 , 353 , 366 , 464 and 710 is substituted the residues have selective substitutions as follows:(i) at positions selected from the group consisting of 47, 82, 142, 205, 236, 252, 255, and 464 and any combination thereof, 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);(ii) at positions selected from the group consisting of 52, 94, 175, 184, 188, 290, 353, and 366, an amino acid residue selected from alanine (A), valine (V), leucine (L), isoleucine (I), proline (P), and methionine (M);(iii) at position 226, an amino acid residue selected from the group consisting of lysine (K), and arginine (R);(iv) at position 78 and 328, an amino acid residue selected from the group consisting of alanine (A), tyrosine (Y), phenylalanine (F), tryptophan (W), and histidine (H),(v) at position 74, an amino acid selected from the group consisting of alanine (A), serine (S), and glutamic acid (E); and(vi) at position 710 of SEQ ID NO:1 a threonine (T) residue;and which the polypeptide comprises the following mutation(s):(a) at position 698 a glycine (G) residue, and/or(b) at position 443 an alanine (A) residue;{'sub': 1', '12, 'wherein the polypeptide catalyzes the conversion of a (C-C)alkane to an alcohol.'}23-. (canceled)4. The polypeptide of claim 1 , comprising ...
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