Method for producing hydroxy-L-pipecolic acid

A method of producing a (5S)-hydroxy-L-pipecolic acid without producing a hydroxy-L-proline, the method comprising:

• allowing an L-pipecolic acid hydroxylase, a microorganism or cell having the ability to produce the enzyme, a processed product of the microorganism or cell, and/or a culture liquid comprising the enzyme and obtained by culturing the microorganism or cell, to act on L-pipecolic acid as a substrate in the presence of 2-oxoglutaric acid and ferrous ion,
• wherein the L-pipecolic acid hydroxylase has properties (1) and (2) below:
• (1) the enzyme can act on L-pipecolic acid in the presence of 2-oxoglutaric acid and ferrous ion to add a hydroxy group to the carbon atom at positions 3, 4, and/or 5 of L-pipecolic acid;
• and
• (2) the enzyme has a catalytic efficiency (kcat/Km) with L-proline that is equal to or less than 7 times the catalytic efficiency (kcat/Km) with L-pipecolic acid,
• wherein the content of L-proline in the L-pipecolic acid as the substrate is not more than 10% (w/w), and
• wherein the L-pipecolic acid hydroxylase comprises one or more proteins selected from (A), (B), and (C) below:
• (A) a protein consisting of the amino acid sequence of SEQ ID NO:
• 2;
• (B) a protein having the same amino acid sequence as the amino acid sequence of SEQ ID NO:
• 2, except that 1 to 10 amino acids are deleted, substituted, inserted, and/or added, which protein has L-pipecolic acid hydroxylase activity;
• and
• (C) a protein having an amino acid sequence with an identity of not less than 90% to the amino acid sequence of SEQ ID NO:
• 2, which protein has the L-pipecolic acid hydroxylase activity.