Baicalein- and Scutellarein- Synthesizing Microorganism, Preparation Method and Applications Thereof
AssigneeCas Center for Excellence In Molecular Plant Sciences
InventorsYong Wang, Jianhua Li, Chenfei TIAN
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Abstract
Provided are a baicalein- and scutellarein-synthesizing microorganism, a preparation method for same, and applications thereof. By modifying a heterologous metabolic pathway of a host cell per a genetic engineering method, acquired is an engineered strain providing a high yield of baicalein and scutellarein. Also provided is a process for utilizing the engineered strain to produce baicalein and scutellarein.
Claims23
A method of producing baicalein and scutellarein, comprising:
- (1) introducing genes expressing flavone 6-hydroxylase and cytochrome P450 oxidoreductase, as well as genes for synthesizing chrysin or apigenin, into a host cell;
- (2) culturing the host cell in a culture system containing phenylalanine and/or tyrosine to produce baicalein or scutellarein.
A method of producing baicalein and scutellarein, comprising:
- (1) introducing genes expressing flavone 6-hydroxylase and cytochrome P450 oxidoreductase into a host cell to obtain a recombinant strain;
- (2) culturing the recombinant host cell in a culture system containing chrysin or apigenin to produce baicalein or scutellarein.
A method for converting chrysin or apigenin into baicalein or scutellarein, comprising:
- catalyzing chrysin or apigenin by flavone 6-hydroxylase and cytochrome P450 oxidoreductase, thereby adding a hydroxyl group to the structure of chrysin or apigenin to form baicalein or scutellarein.
A recombinant host cell comprising exogenous genes expressing flavone 6-hydroxylase and cytochrome P450 oxidoreductase.
(canceled)
A method of preparing a host cell for producing baicalein and scutellarein, comprising:
- introducing genes expressing flavone 6-hydroxylase and cytochrome P450 oxidoreductase into the host cell to obtain a recombinant strain;
- preferably, the method also comprises:
- introducing genes for synthesizing chrysin or apigenin.
A mutant flavonoid 6-hydroxylase, which corresponds to the wild-type flavonoid 6-hydroxylase but the N-terminal amino acids (1-10) to (20-30) are truncated;
- preferably, the N-terminal amino acids (2-5) to (22-28) are truncated;
- more preferably, the mutant flavonoid 6-hydroxylase has the amino acid sequence shown in SEQ ID NO:
- 2.
A mutant cytochrome P450 oxidoreductase, which corresponds to the wild-type cytochrome P450 oxidoreductase but the N-terminal amino acids (1-20) to (60-85) are truncated;
- preferably, the N-terminal amino acids (2-10) to (65-80) are truncated;
- more preferably, the N-terminal amino acids (2-5) to (70-75) are truncated;
- more preferably, the mutant cytochrome P450 oxidoreductase has the amino acid sequence shown in SEQ ID NO:
- 8.
A polynucleotide, encoding:
- a mutant flavone 6-hydroxylase having N-terminal amino acids (1-10) to (20-30) or (2-5) to (22-28) truncated from wild-type flavonoid 6-hydroxylase;
- wherein preferably the mutant flavonoid 6-hydroxylase has the amino acid sequence of SEQ ID NO:
- 2;
- or
- a mutant cytochrome P450 oxidoreductase having N-terminal amino acids (1-20) to (60-85), or (2-10) to (65-80), or (2-5) to (70-75) truncated from wild-type cytochrome P450 oxidoreductase;
- wherein preferably the mutant cytochrome P450 oxidoreductase has the amino acid sequence of SEQ ID NO:
- 8;
- or
- a fusion polypeptide comprising a mutant flavone 6-hydroxylase fused with a peptide tag, the peptide tag being selected from the group consisting of 8RP, Sumo, MBP, and 2B1;
- wherein preferably the peptide tag is MBP or 2B1.
(canceled)
Dependencies are inferred from the claim text.