Cell-free bioproduction of b-cryptoxanthin and zeaxanthin
US-2024368663-A1 · Nov 7, 2024 · US
US9926539B2 · US · B2
| Field | Value |
|---|---|
| Publication number | US-9926539-B2 |
| Application number | US-201615006221-A |
| Country | US |
| Kind code | B2 |
| Filing date | Jan 26, 2016 |
| Priority date | Apr 15, 2013 |
| Publication date | Mar 27, 2018 |
| Grant date | Mar 27, 2018 |
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Two novel cytochrome P450 genes are isolated from sorghum, each gene encoding a protein having pentadecatrienyl resorcinol hydroxylase activity. Expression vectors containing these sequences are made and used to elevate levels of pentadecatrienyl resorcinol hydroxylase in transgenic cells and organisms.
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We, the inventors, claim: 1. An expression vector comprising a promoter and a heterologous polynucleotide, wherein said heterologous polynucleotide encodes a pentadecatrienyl resorcinol hydroxylase having the amino acid sequence of SEQ ID NO: 14, wherein said promoter is operatively linked to said heterologous polynucleotide, and wherein said promoter is selected from the group consisting of a constitutive promoter, a tissue-specific promoter, and an inducible promoter. 2. The expression vector of claim 1 , wherein said heterologous polynucleotide comprises SEQ ID NO: 13. 3. A transformed cell capable of producing pentadecatrienyl resorcinol hydroxylase, said transformed cell comprising the expression vector of claim 1 , wherein said transformed cell is selected from the group consisting of a plant cell, a fungus, a blue-green algae, and a bacterium. 4. The transformed cell of claim 3 , wherein said heterologous polynucleotide comprises SEQ ID NO: 13. 5. A transgenic organism capable of producing pentadecatrienyl resorcinol hydroxylase, said transgenic organism comprising the transformed cell of claim 3 ; wherein said transgenic organism produces elevated levels of pentadecatrienyl resorcinol hydroxylase compared to the levels of pentadecatrienyl resorcinol hydroxylase produced by the non-transformed organism. 6. The transgenic organism of claim 5 , wherein said heterologous polynucleotide comprises SEQ ID NO: 13. 7. A method of manipulating pentadecatrienyl resorcinol hydroxylase levels in a transformed cell or transformed organism, said method comprising introducing into a wild-type cell or a wild-type organism an expression vector comprising a heterologous polynucleotide and a promoter to produce said transformed cell or said transformed organism, wherein said heterologous polynucleotide is operably linked to said promoter, wherein said promoter is active in said cell or organism, wherein said promoter is selected from the group consisting of a constitutive promoter, a tissue-specific promoter, and an inducible promoter, wherein said heterologous polynucleotide encodes a polypeptide having the amino acid sequence of SEQ ID NO: 14, wherein said polypeptide has pentadecatrienyl resorcinol hydroxylase activity, wherein said transformed cell or transformed organism is selected from the group consisting of a plant, a fungus, a blue-green algae, and a bacterium, and allowing the production of pentadecatrienyl resorcinol hydroxylase in said transformed cell or transformed organism. 8. The transformed plant cell or plant made by the method of claim 7 , or progeny thereof, wherein said transformed plant cell or plant or progeny thereof is selected for having increased levels of pentadecatrienyl resorcinol hydroxylase as compared to the level of pentadecatrienyl resorcinol hydroxylase present in the non-transformed plant cell or plant of the same variety.
with NADH or NADPH as one donor, and incorporation of one atom of oxygen (1.14.13) · CPC title
inducible or controllable · CPC title
containing the quinoid structure · CPC title
aromatic · CPC title
tissue or cell-specific · CPC title
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