Method of producing (-)-rotundone
AssigneeT. Hasegawa Co., Ltd.
InventorsKuniki KINO, Toshiki FURUYA
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Abstract
The present invention provides a method of producing (−)-rotundone from α-guaiene. The method includes the steps of: (1) allowing a cytochrome P450 protein to act on α-guaiene, which cytochrome P450 belongs to the CYP152 family and is capable of oxidizing the methylene group at position 3 of α-guaiene to the carbonyl group; and/or (2) allowing a cytochrome P450 protein to act on α-guaiene in the presence of an electron transfer protein capable of transferring electrons to the cytochrome P450 protein, which cytochrome P450 belongs to the CYP152, CYP106, or CYP107 family and is capable of oxidizing the methylene group at position 3 of α-guaiene to the carbonyl group.
Claims11
A method of producing (−)-rotundone from α-guaiene, the method comprising the following steps (1) and/or (2):
- (1) allowing a cytochrome P450 protein to act on α-guaiene, which cytochrome P450 protein belongs to a CYP152 family and is capable of oxidizing the methylene group at position 3 of α-guaiene to the carbonyl group;
- (2) allowing a cytochrome P450 protein to act on α-guaiene in the presence of an electron transfer protein capable of transferring electrons to the cytochrome P450 protein, which cytochrome P450 protein belongs to a CYP152, CYP106, or CYP107 family and is capable of oxidizing the methylene group at position 3 of α-guaiene to the carbonyl group.
A recombinant vector for converting α-guaiene to (−)-rotundone, the recombinant vector comprising a DNA selected from:
- (g) a DNA having a nucleotide sequence of SEQ ID NO:
- 5;
- (h) a DNA that has a nucleotide sequence having a sequence identity of at least 90% to a nucleotide sequence of SEQ ID NO:
- 5;
- (i) a DNA hybridizable, under stringent conditions, with a DNA having a nucleotide sequence complementary to a nucleotide sequence of SEQ ID NO:
- 5;
- (j) a DNA encoding a protein that has an amino acid sequence having a sequence identity of at least 90% to an amino acid sequence of SEQ ID NO:
- 1;
- (k) a DNA encoding a protein that has an amino acid sequence including a conservative substitution of at least one amino acid in an amino acid sequence of SEQ ID NO:
- 1 and having a sequence identity of at least 80% to the amino acid sequence of SEQ ID NO:
- 1;
- (l) a DNA having a nucleotide sequence of any one of SEQ ID NOS:
- 6 to 8;
- (m) a DNA that has a nucleotide sequence having a sequence identity of at least 90% to a nucleotide sequence of any one of SEQ ID NOS:
- 6 to 8;
- (n) a DNA hybridizable, under stringent conditions, with a DNA having a nucleotide sequence complementary to a nucleotide sequence of any one of SEQ ID NOS:
- 6 to 8;
- (o) a DNA encoding a protein that has an amino acid sequence having a sequence identity of at least 90% to an amino acid sequence of any one of SEQ ID NOS:
- 2 to 4;
- and
- (p) a DNA encoding a protein that has an amino acid sequence including a conservative substitution of at least one amino acid in an amino acid sequence of any one of SEQ ID NOS:
- 2 to 4 and having a sequence identity of at least 80% to the amino acid sequence of any one of SEQ ID NOS:
- 2 to 4.
A recombinant vector for converting α-guaiene to (−)-rotundone, the recombinant vector comprising:
- a DNA selected from (g), (h), (i), (j), (k), (l), (m), (n), (o), and (p), and
- a DNA selected from (G), (H), and (I),
- wherein (g), (h), (i), (j), (k), (l), (m), (n), (o), (p), (G), (H), and (I) are as follows:
- (g) a DNA having a nucleotide sequence of SEQ ID NO:
- 5;
- (h) a DNA that has a nucleotide sequence having a sequence identity of at least 90% to a nucleotide sequence of SEQ ID NO:
- 5;
- (i) a DNA hybridizable, under stringent conditions, with a DNA having a nucleotide sequence complementary to a nucleotide sequence of SEQ ID NO:
- 5;
- (j) a DNA encoding a protein that has an amino acid sequence having a sequence identity of at least 90% to an amino acid sequence of SEQ ID NO:
- 1;
- (k) a DNA encoding a protein that has an amino acid sequence including a conservative substitution of at least one amino acid in an amino acid sequence of SEQ ID NO:
- 1 and having a sequence identity of at least 80% to the amino acid sequence of SEQ ID NO:
- 1;
- (l) a DNA having a nucleotide sequence of any one of SEQ ID NOS:
- 6 to 8;
- (m) a DNA that has a nucleotide sequence having a sequence identity of at least 90% to a nucleotide sequence of any one of SEQ ID NOS:
- 6 to 8;
- (n) a DNA hybridizable, under stringent conditions, with a DNA having a nucleotide sequence complementary to a nucleotide sequence of any one of SEQ ID NOS:
- 6 to 8;
- (o) a DNA encoding a protein that has an amino acid sequence having a sequence identity of at least 90% to an amino acid sequence of any one of SEQ ID NOS:
- 2 to 4;
- (p) a DNA encoding a protein that has an amino acid sequence including a conservative substitution of at least one amino acid in an amino acid sequence of any one of SEQ ID NOS:
- 2 to 4 and having a sequence identity of at least 80% to the amino acid sequence of any one of SEQ ID NOS:
- 2 to 4,
- (G) a DNA having a nucleotide sequence of SEQ ID NO:
- 9,
- (H) a DNA that has a nucleotide sequence having a sequence identity of at least 90% to a nucleotide sequence of SEQ ID NO:
- 9,
- (I) a DNA hybridizable, under stringent conditions, with a DNA having a nucleotide sequence complementary to a nucleotide sequence of SEQ ID NO:
- 9.
Dependencies are inferred from the claim text.