# US 2010/0170010 A1 · Use of genes encoding membrane transporter pumps to stimulate the production of secondary metabolites in biological cells > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 2010/0170010 A1 - Document ID: US20100170010A1 - Status: Published application - Kind: A1 - Application number: 12/592,505 - Filed: 2009-11-24 - Published: 2010-07-01 - Earliest claimed: 2001-04-18 - Inventors: Alain Goossens; Dirk G Inzé - Web page: https://patentgrep.com/patent/US20100170010A1 ## CPC - C07K 14/395: from Saccharomyces - C12N 15/8242: {with non-agronomic quality (output) traits, e.g. for industrial processing; Value added, non-agronomic traits} - C07K: PEPTIDES (peptides in foodstuffs A23; obtaining protein compositions for foodstuffs, working-up proteins for foodstuffs A23J; preparations for medicinal purposes A61K; peptides containing beta-lactam rings C07D; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, C07D; ergot alkaloids of the cyclic peptide type C07D519/02; macromolecular compounds having statistically distributed amino acid units in their molecules, i.e. when the preparation does not provide for a specific; but for a random sequence of the amino acid units, homopolyamides and block copolyamides derived from amino acids C08G69/00; macromolecular products derived from proteins C08H1/00; preparation of glue or gelatine C09H; single cell proteins, enzymes C12N; genetic engineering processes for obtaining peptides C12N15/00; compositions for measuring or testing processes involving enzymes C12Q; investigation or analysis of biological material G01N33/00) - C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00) ## Abstract The invention relates to the field of secondary metabolite production in plants and plant cell cultures. More specifically, the invention relates to the use of transporters and more particularly ABC-transporters to enhance the production and/or secretion of secondary metabolites in plants and plant cell cultures. ## Claims ### Claim 1 A method of inducing or enhancing production of at least one secondary metabolite by plant cells, said method comprising: transforming plant cells with an expression vector comprising an expression cassette comprising a gene encoding an ABC-transporter; wherein said ABC-transporter comprises a Walker A box, a Walker B box, and a Nucleotide Binding Fold; wherein said ABC-transporter functions to transport at least one secondary metabolite in plant cells; selecting transformed plant cells having an induced or enhanced production of at least one secondary metabolite; and propagating such selected transformed plant cells. ### Claim 2 (depends on 1) The method according to claim 1, wherein said gene encoding an ABC transporter comprises a polynucleotide sequence having at least 91% identity to the polynucleotide sequence of SEQ ID NO:17. ### Claim 3 (depends on 1) The method according to claim 1 wherein the secondary metabolites are alkaloids. ### Claim 4 (depends on 1) The method according to claim 1 wherein the ABC-transporters are of plant, fungal, or mammalian origin. ### Claim 5 (depends on 1) The method according to claim 1 wherein the induction or enhancement of the production of at least one secondary metabolite by plant cells results from enhancing the transport of said secondary metabolite into a vacuole. ### Claim 6 (depends on 5) The method according to claim 5 wherein the secondary metabolites are alkaloids. ### Claim 7 (depends on 5) The method according to claim 5 wherein the ABC-transporters are of plant, fungal, or mammalian origin. ### Claim 8 (depends on 1) The method according to claim 1 wherein the induction or enhancement of the production of at least one secondary metabolite by plant cells results from enhancing the transport of said secondary metabolite to the extracellular space of the plant cell. ### Claim 9 (depends on 5) The method according to claim 5 wherein the secondary metabolites are alkaloids. ### Claim 10 (depends on 5) The method according to claim 5 wherein the ABC-transporters are of plant, fungal, or mammalian origin. ### Claim 11 A method of stimulating the production of secondary metabolites by plants, the method comprising: transforming said plants with an expression vector comprising an expression cassette comprising a gene encoding an ABC-transporter; wherein said ABC-transporter comprises a Walker A box, a Walker B box, and a Nucleotide Binding Fold; and wherein said ABC-transporter functions to transport at least one secondary metabolite in plant cells; selecting transformed plants based upon enhanced production of secondary metabolites; and propagating such selected transformed plants. ### Claim 12 (depends on 11) The method according to claim 11, wherein said gene encoding an ABC transporter comprises a polynucleotide sequence having at least 91% identity to the polynucleotide sequence of SEQ ID NO:17. ### Claim 13 (depends on 11) The method according to claim 11 wherein the secondary metabolites are alkaloids. ### Claim 14 (depends on 11) The method according to claim 11 wherein the ABC-transporters are of plant, fungal, or mammalian origin. ### Claim 15 A transgenic plant cell culture displaying an enhanced production of at least one secondary metabolite, wherein said transgenic plant cell is transformed with an expression vector comprising an expression cassette comprising a gene encoding an ABC-transporter; wherein said ABC-transporter comprises a Walker A box, a Walker B box, and a Nucleotide Binding Fold; and wherein said ABC-transporter functions to transport at least one secondary metabolite in plant cells. ### Claim 16 (depends on 15) The method according to claim 15, wherein said gene encoding an ABC transporter comprises a polynucleotide sequence having at least 91% identity to the polynucleotide sequence of SEQ ID NO:17. ### Claim 17 (depends on 15) The transgenic plant cell culture of claim 15 further characterized in having (1) an increased vacuolar localization of said at least one secondary metabolite, or (2) a secretion or an increased secretion of said at least one secondary metabolite. ### Claim 18 A transgenic plant material selected from the group consisting of a plant, plant cells, plant seeds and plant progeny, said transgenic plant material capable of an enhanced production of at least one secondary metabolite, said transgenic plant material transformed with an expression vector comprising an expression cassette comprising a gene encoding an ABC-transporter; wherein said ABC-transporter comprises a Walker A box, a Walker B box, and a Nucleotide Binding Fold; and wherein said ABC-transporter functions to transport at least one secondary metabolite in plant cells. ### Claim 19 (depends on 18) The transgenic plant material of claim 18, wherein said gene encoding an ABC transporter comprises a polynucleotide sequence having at least 91% identity to the polynucleotide sequence of SEQ ID NO:17. ### Claim 20 (depends on 18) The transgenic plant material of claim 18 further characterized in having an increased vacuolar localization of said at least one secondary metabolite. ### Claim 21 An isolated polynucleotide sequence comprising a sequence having at least 91% identity to the polynucleotide sequence of SEQ ID NO:17; wherein the isolated polynucleotide sequence induces or enhances production of at least one secondary metabolite in plants. ### Claim 22 A process for producing a plant cell exhibiting an enhanced production of at least one secondary metabolite, said process comprising: transforming a plant cell with an expression cassette comprising a gene encoding an ABC-transporter; wherein said ABC-transporter comprises a Walker A box, a Walker B box, and a Nucleotide Binding Fold; wherein said ABC-transporter functions to transport at least one secondary metabolite in plant cells; and selecting transformed plant cells exhibiting enhanced transport of said at least one secondary metabolite into a vacuole. ### Claim 23 (depends on 22) The process according to claim 22, wherein said gene encoding an ABC transporter comprises a polynucleotide sequence having at least 91% identity to the polynucleotide sequence of SEQ ID NO:17. ### Claim 24 (depends on 22) A plant cell produced by the process of claim 22. ### Claim 25 (depends on 24) A transgenic plant including the plant cell of claim 24. ### Claim 26 An isolated polynucleotide useful for producing a plant cell exhibiting an enhanced production of at least one secondary metabolite, said isolated polynucleotide comprising: a first sequence of nucleotide bases constituting a means for inducing or enhancing production of at least one secondary metabolite in plants or plant cells, and a second sequence of nucleotides bases, operatively positioned with respect to said first sequence, constituting a means for promoting expression of said first sequence. ### Claim 27 (depends on 26) The isolated polynucleotide sequence of claim 26, wherein the isolated polynucleotide sequence comprises a polynucleotide sequence having at least 91% identity to the polynucleotide sequence of SEQ ID NO:17. ### Claim 28 A method of inducing or enhancing production or cellular secretion of at least one endogenous secondary metabolite by a plant cell, the method comprising: transforming the plant cell with an expression vector comprising an expression cassette comprising a gene encoding an ABC-transporter, wherein said ABC-transporter comprises a Walker A box, a Walker B box, and a Nucleotide Binding Fold, and functions to transport at least one secondary metabolite in plant cells; wherein the secondary metabolite is an endogenous metabolic product of the plant cell, and is transported from the cell to the extracellular space; wherein the amount of secondary metabolite recoverable from the cell is increased; selecting a transformed plant cell having an induced or enhanced production of at least one secondary metabolite; and propagating such selected transformed plant cell ### Claim 29 (depends on 28) The method according to claim 28, wherein said gene encoding an ABC transporter comprises a polynucleotide sequence having at least 91% identity to the polynucleotide sequence of SEQ ID NO:17. ### Claim 30 (depends on 28) The method according to claim 28 wherein the secondary metabolite is an alkaloid. ### Claim 31 (depends on 28) The method according to claim 28 wherein the ABC-transporter is of plant, fungal, or mammalian origin.