Heterologous production of patchoulol, beta-santalene, and sclareol in moss cells
AssigneesUniversity of Copenhagen, Firmenich SA
InventorsHenrik SIMONSEN, Dong-Fang CHEN, Xin ZHAN, Xiwu PAN, Brian Christopher KING
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Abstract
The present invention relates to methods for preparing patchoulol, β-santalene, and sclareol in transgenic moss cells that include heterologous nucleic acid molecules encoding a polypeptide or synthase capable of using FPP or GGPP as a substrate. Methods for producing the transgenic moss cell, as well as the transgenic moss cell itself are also disclosed.
Claims10
A method for producing a transgenic moss cell comprising introducing into a moss cell capable of producing farnesyl-pyrophosphate (FPP) or geranylgeranyl-pyrophosphate (GGPP) a heterologous nucleic acid molecule encoding a polypeptide, the polypeptide having a patchoulol synthase activity or β-santalene synthase activity to catalyze the conversion of FPP to patchoulol or β-santalene, or a sclareol synthase activity to catalyse the conversion of GGPP to sclareol, wherein the heterologous nucleic acid molecule is at least 70% identical to SEQ ID NO 1, SEQ ID NO 2, or SEQ ID NO 3.
A method for preparing patchoulol, β-santalene, or sclareol in a transgenic moss cell comprising;
- a) introducing into a moss cell capable of producing farnesyl-pyrophosphate (FPP) or geranylgeranyl-pyrophosphate (GGPP) a heterologous nucleic acid molecule encoding a polypeptide, the polypeptide having a patchoulol synthase activity or β-santalene synthase activity to catalyze the conversion of FPP to patchoulol or β-santalene, or a sclareol synthase activity to catalyse the conversion of GGPP to sclareol, wherein the heterologous nucleic acid molecule is at least 70% identical to SEQ ID NO 1, SEQ ID NO 2, or SEQ ID NO 3,
- b) culturing the transgenic moss cell to express or overexpress the polypeptide encoded by the heterologous nucleic acid molecule, and
- c) isolating patchoulol, β-santalene, and/or sclareol produced in step b).
A transgenic moss cell capable of producing FPP or GGPP, the transgenic moss cell comprises a heterologous nucleic acid molecule encoding one or more polypeptides, the polypeptide having a patchoulol synthase activity or β-santalene synthase activity to catalyze the conversion of FPP to patchoulol or β-santalene, or a sclareol synthase activity to catalyse the conversion of GGPP to sclareol, wherein the heterologous nucleic acid molecule is at least 70% identical to SEQ ID NO 1, SEQ ID NO 2, or SEQ ID NO 3.
Dependencies are inferred from the claim text.