Method for producing steviol and steviol glycoside using aobgl3 homolog
AssigneeSuntory Holdings Limited
InventorsMisa OCHIAI, Eiichiro ONO
Life of this patent
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Abstract
The present invention provides a method for preparing a steviol glycoside and/or steviol comprising reacting glycosidase AOBGL1 and/or AOBGL3 or a variant thereof with a steviol glycoside having at least one monoglucoside bond and/or monoglucosyl ester bond, thereby cleaving said monoglucoside bond and/or monoglucosyl ester bond.
Claims13
A method for preparing a steviol glycoside and/or steviol comprising reacting a protein selected from the group consisting of proteins (a) to (c) shown below with a steviol glycoside having at least one monoglucoside bond and/or monoglucosyl ester bond, thereby cleaving said monoglucoside bond and/or monoglucosyl ester bond:
- (a) a protein consisting of the amino acid sequence of SEQ ID NO:
- 3 or 4;
- (b) a protein consisting of an amino acid sequence wherein 1 to 77 amino acids have been deleted, substituted, inserted, and/or added in the amino acid sequence of SEQ ID NO:
- 3 or 4, and having an activity to cleave a monoglucoside bond and/or monoglucosyl ester bond;
- and
- (c) a protein having an amino acid sequence having 90% or more sequence identity to the amino acid sequence of SEQ ID NO:
- 3 or 4, and having an activity to cleave a monoglucoside bond and/or monoglucosyl ester bond of a steviol glycoside.
A method for producing a steviol glycoside and/or steviol comprising culturing a non-human transformant obtained by introducing a polynucleotide selected from the group consisting of polynucleotides (a) to (e) shown below into a host producing a steviol glycoside having at least one monoglucoside bond and/or monoglucosyl ester bond:
- (a) a polynucleotide consisting of the nucleotide sequence of SEQ ID NO:
- 1;
- (b) a polynucleotide encoding a protein consisting of the amino acid sequence of SEQ ID NO:
- 3 or 4;
- (c) a polynucleotide encoding a protein consisting of an amino acid sequence wherein 1 to 77 amino acids have been deleted, substituted, inserted, and/or added in the amino acid sequence of SEQ ID NO:
- 3 or 4, and having an activity to cleave a monoglucoside bond and/or monoglucosyl ester bond of a steviol glycoside;
- (d) a polynucleotide encoding a protein having an amino acid sequence having 90% or more sequence identity to the amino acid sequence of SEQ ID NO:
- 3 or 4, and having an activity to cleave a monoglucoside bond and/or monoglucosyl ester bond of a steviol glycoside;
- and
- (e) a polynucleotide which hybridizes under highly stringent conditions to a polynucleotide consisting of a nucleotide sequence complementary to a polynucleotide consisting of the nucleotide sequence of SEQ ID NO:
- 1 and which encodes a protein having an activity to cleave a monoglucoside bond and/or monoglucosyl ester bond of a steviol glycoside.
A method for preparing a steviol glycoside and/or steviol comprising contacting an enzyme agent derived from a non-human transformed cell obtained by introducing, into a host cell, a polynucleotide selected from the group consisting of polynucleotides (a) to (e) shown below, with a steviol glycoside having at least one monoglucoside bond and/or monoglucosyl ester bond, thereby cleaving said monoglucoside bond and/or monoglucosyl ester bond:
- (a) a polynucleotide consisting of the nucleotide sequence of SEQ ID NO:
- 1;
- (b) a polynucleotide encoding a protein consisting of the amino acid sequence of SEQ ID NO:
- 3 or 4;
- (c) a polynucleotide encoding a protein consisting of an amino acid sequence wherein 1 to 77 amino acids have been deleted, substituted, inserted, and/or added in the amino acid sequence of SEQ ID NO:
- 3 or 4, and having an activity to cleave a monoglucoside bond and/or monoglucosyl ester bond of a steviol glycoside;
- (d) a polynucleotide encoding a protein having an amino acid sequence having 90% or more sequence identity to the amino acid sequence of SEQ ID NO:
- 3 or 4, and having an activity to cleave a monoglucoside bond and/or monoglucosyl ester bond of a steviol glycoside;
- and
- (e) a polynucleotide which hybridizes under highly stringent conditions to a polynucleotide consisting of a nucleotide sequence complementary to a polynucleotide consisting of the nucleotide sequence of SEQ ID NO:
- 1 and which encodes a protein having an activity to cleave a monoglucoside bond and/or monoglucosyl ester bond of a steviol glycoside.
A method for producing a steviol glycoside and/or steviol comprising reacting a protein selected from the group consisting of proteins (a) to (c) shown below, a protein selected from the group consisting of proteins (d) to (f) shown below, and a steviol glycoside having at least one monoglucoside bond and/or monoglucosyl ester bond:
- (a) a protein consisting of the amino acid sequence of SEQ ID NO:
- 3 or 4;
- (b) a protein consisting of an amino acid sequence wherein 1 to 77 amino acids have been deleted, substituted, inserted, and/or added in the amino acid sequence of SEQ ID NO:
- 3 or 4, and having an activity to cleave a monoglucoside bond and/or monoglucosyl ester bond of a steviol glycoside;
- (c) a protein having an amino acid sequence having 90% or more sequence identity to the amino acid sequence of SEQ ID NO:
- 3 or 4, and having an activity to cleave a monoglucoside bond and/or monoglucosyl ester bond of a steviol glycoside;
- (d) a protein consisting of the amino acid sequence of SEQ ID NO:
- 7 or 8;
- (e) a protein consisting of an amino acid sequence wherein 1 to 77 amino acids have been deleted, substituted, inserted, and/or added in the amino acid sequence of SEQ ID NO:
- 7 or 8, and having an activity to cleave an unbranched β-1,2-glucoside bond of a steviol glycoside;
- and
- (f) a protein having an amino acid sequence having 90% or more sequence identity to the amino acid sequence of SEQ ID NO:
- 7 or 8, and having an activity to cleave an unbranched β-1,2-glucoside bond of a steviol glycoside.
Dependencies are inferred from the claim text.