Enzymatic methods for converting lca and 3-kca to udca and 3-kudca
AssigneeSandhill One, LLC
InventorsJ. GREGORY REID, JAYACHANDRA P. REDDY, MATT GREGORY, URSULA SCHELL, BERNHARD J. PAUL
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Abstract
7β-hydroxylation systems are provided, as well as methods for producing ?P-hydroxy derivatives of lithocholic acid and 3-keto-lithocholic acid from such systems. Also provided are recombinant organisms useful for the production of such enzymatic systems, and to plasmids that encode for such enzymes.
Claims32
1) A method of converting LCA or 3-KCA, or a carboxylic acid ester, carboxylic amide, or carboxylate salt thereof, to UDCA or 3-KUDCA, or a carboxylic acid ester, carboxylic amide, or carboxylate salt thereof, comprising contacting the LCA or 3-KCA, or carboxylic acid ester, carboxylic amide, or carboxylate salt thereof, with a 7β-hydroxylase system in the presence of a yeast, or an extract or lysate thereof, wherein the 7β-hydroxylase system is not native to the yeast.
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19) A plasmid comprising a nucleic acid sequence selected from SEQ ID NO. 8;
- SEQ ID NO. 11;
- SEQ ID NO. 14;
- SEQ ID NO. 17;
- SEQ ID NO. 20;
- SEQ ID NO. 23;
- SEQ ID NO. 26;
- SEQ ID NO. 29;
- or SEQ ID NO. 32;
- or a nucleic acid sequence having at least 85%, 90%, 95%, 98%, or 99%, identity with any of the foregoing sequences.
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24) An organism transformed by a CYP encoding nucleic acid sequence selected from SEQ ID NO. 8;
- SEQ ID NO. 11;
- SEQ ID NO. 14;
- SEQ ID NO. 17;
- SEQ ID NO. 20;
- SEQ ID NO. 23;
- SEQ ID NO. 26;
- SEQ ID NO. 29;
- and SEQ ID NO. 32;
- or a nucleic acid sequence having at least 85%, 90%, 95%, 98%, or 99%, identity with any of the foregoing nucleic acid sequences.
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Dependencies are inferred from the claim text.