Patent ID: 7531335

Claim:
A process for the enzymatic production of subterminally hydroxylated aliphatic carboxylic acids, which comprises a1) culturing a recombinant microorganism which has been transformed with a vector which encompasses an expression construct comprising, under the genetic control of regulatory nucleic acid sequences, a sequence which encompasses a nucleic acid sequence encoding a modified monooxygenase having a modification in the amino acid sequence of SEQ ID NO:2, which modification consists of one functional mutation in each of amino acid sequence positions 87 and 188 and, optionally, at least one additional functional mutation in one of amino acid sequence positions 26, 47, 72, 74 and 354, wherein: Phe 87 is replaced by at Val, Ala or Leu; Leu 188 is replaced by Asn, Gln, Arg, Lys, Ala, Gly, Ser or Trp; Ala 74 is replaced by Val or Gly; Arg 47 is replaced by His, Tyr or Phe; Val 26 is replaced by Ser or Thr; Ser 72 is replaced by Ala, Leu, Ile or Gly; or, Met 354 is replaced by Ser or Thr, wherein the functional mutation in comparison with the wild-type enzyme, results in an altered activity or regioselectivity in the subterminal enzymatic hydroxylation of an aliphatic C 8 -C 12 -carboxylic acid, whereby culturing is performed in the presence of a culture medium which contains at least one hydroxylatable C 8 -C 12 -carboxylic acid or a derivative thereof, said derivative being selected from an alkyl ester, an amide or an anhydride of the at least one hydroxylatable C 8 -C 12 -carboxylic acid; or a2) incubating a reaction medium containing at least one hydroxylatable C 8 -C 12 -carboxylic acid or a derivative thereof, said derivative being selected from an alkyl ester, an amide or an anhydride of the at least one hydroxylatable C 9 -C 12 -carboxylic acid with a modified monooxygenase as defined above, and b) isolating the resulting hydroxylated products from the medium.