| CPC C12N 9/93 (2013.01) [C12N 9/0004 (2013.01); C12N 9/0006 (2013.01); C12P 19/60 (2013.01)] | 15 Claims |
|
1. A synthesis method for a betalain pigment, comprising:
culturing a host that has introduced therein the following gene encoding an amaranthin synthetase or gomphrenin-I-glucuronide synthetase, a gene encoding an enzyme having activity of hydroxylating a 3-position of a phenol ring of tyrosine, a gene encoding an enzyme having L-DOPA oxidase activity, a gene encoding an enzyme having activity of glycosylating a phenolic hydroxy group, and a gene encoding an enzyme having DOPA 4,5-dioxygenase activity, and that has an ability to produce tyrosine or 3-hydroxy-L-tyrosine (L-DOPA), and extracting a betalain pigment from the host after the culturing, wherein the gene encoding an enzyme having activity of hydroxylating a 3-position of a phenol ring of tyrosine and the gene encoding an enzyme having L-DOPA oxidase activity is a single gene or different genes; or
culturing a host that has introduced therein the following gene encoding an amaranthin synthetase or gomphrenin-I-glucuronide synthetase, a gene encoding an enzyme having activity of hydroxylating a 3-position of a phenol ring of tyrosine, a gene encoding an enzyme having L-DOPA oxidase activity, a gene encoding a betanidin-to-betanin synthetase, and a gene encoding an enzyme having DOPA 4,5-dioxygenase activity, and that has an ability to produce tyrosine or 3-hydroxy-L-tyrosine, and extracting a betalain pigment from the host after the culturing, wherein the gene encoding an enzyme having activity of hydroxylating a 3-position of a phenol ring of tyrosine and the gene encoding an enzyme having L-DOPA oxidase activity is a single gene or different genes; or
culturing a host that has introduced therein the following gene encoding an amaranthin synthetase or gomphrenin-I-glucuronide synthetase, a gene encoding an enzyme having activity of hydroxylating a 3-position of a phenol ring of tyrosine, a gene encoding an enzyme having L-DOPA oxidase activity, a gene encoding a betanidin-to-gomphrenin-I (betanidin 6-O-glucoside) synthetase, and a gene encoding an enzyme having DOPA 4,5-dioxygenase activity, and that has an ability to produce tyrosine or 3-hydroxy-L-tyrosine, and extracting a betalain pigment from the host after the culturing, wherein the gene encoding an enzyme having activity of hydroxylating a 3-position of a phenol ring of tyrosine and the gene encoding an enzyme having L-DOPA oxidase activity is a single gene or different genes; or
culturing a host that has introduced therein the following gene encoding an amaranthin synthetase or gomphrenin-I-glucuronide synthetase, and that has enzyme activity of having activity of hydroxylating a 3-position of a phenol ring of tyrosine, enzyme activity of having L-DOPA oxidase activity, enzyme activity of having activity of glycosylating a phenolic hydroxy group, enzyme activity of having DOPA 4,5-dioxygenase activity, and an ability to produce tyrosine or 3-hydroxy-L-tyrosine, and extracting a betalain pigment from the host after the culturing; or
culturing a host that has introduced therein the following gene encoding an amaranthin synthetase or gomphrenin-I-glucuronide synthetase, and that has enzyme activity of having activity of hydroxylating a 3-position of a phenol ring of tyrosine, enzyme activity of having L-DOPA oxidase activity, enzyme activity of having betanidin-to-betanin synthesis activity, enzyme activity of having DOPA 4,5-dioxygenase activity, and an ability to produce tyrosine or 3-hydroxy-L-tyrosine, and extracting a betalain pigment from the host after the culturing; or
culturing a host that has introduced therein the following gene encoding an amaranthin synthetase or gomphrenin-I-glucuronide synthetase, and that has enzyme activity of having activity of hydroxylating a 3-position of a phenol ring of tyrosine, enzyme activity of having L-DOPA oxidase activity, enzyme activity of having betanidin-to-gomphrenin-I synthesis activity, enzyme activity of having DOPA 4,5-dioxygenase activity, and an ability to produce tyrosine or 3-hydroxy-L-tyrosine, and extracting a betalain pigment from the host after the culturing,
wherein, in the production method for a betalain pigment, the gene encoding an amaranthin synthetase or gomphrenin-I-glucuronide synthetase is any one or more selected from the following:
(1) a gene encoding a polypeptide formed of an amino acid sequence set forth in SEQ ID NO: 2, 4, 6, 8, 10, or 12;
(2) a gene encoding a polypeptide that has 1 to 20 amino acids substituted, deleted, inserted, and/or added in the amino acid sequence set forth in SEQ ID NO: 2, 4, 6, 8, 10, or 12, and that has a substantially equivalent ability to synthesize amaranthin or gomphrenin-I-glucuronide to that of the amino acid sequence set forth in SEQ ID NO: 2, 4, 6, 8, 10, or 12;
(3) a gene encoding a polypeptide that has 90% or more homology to the amino acid sequence set forth in SEQ ID NO: 2, 4, 6, 8, 10, or 12, and that has a substantially equivalent ability to synthesize amaranthin or gomphrenin-I-glucuronide to that of the amino acid sequence set forth in SEQ ID NO: 2, 4, 6, 8, 10, or 12;
(4) a gene formed of DNA formed of a base sequence set forth in SEQ ID NO: 1, 3, 5, 7, 9, or 11;
(5) a gene formed of DNA that hybridizes with DNA formed of a base sequence complementary to DNA formed of the base sequence set forth in SEQ ID NO: 1, 3, 5, 7, 9, or 11 under stringent conditions, and that encodes a polypeptide having an ability to synthesize amaranthin or gomphrenin-I-glucuronide;
(6) a gene formed of DNA having a 1- to 50-base sequence substituted, deleted, inserted, and/or added in DNA formed of the base sequence set forth in SEQ ID NO: 1, 3, 5, 7, 9, or 11; and
(7) a gene formed of DNA having 90% or more homology to DNA formed of the base sequence set forth in SEQ ID NO: 1, 3, 5, 7, 9, or 11.
|