Increased production of ginsenosides through yeast cell organelle improvement

Abstract

Provided are a recombinant yeast having improved ability to produce ginsenoside, which is prepared by overexpressing INO2 and INO4 or deleting OPT1 in a yeast having ability to produce ginsenoside, a method of preparing the yeast, and a method of producing ginsenoside by using the yeast.

What is claimed is: 
     
         1 . A recombinant yeast for producing ginsenoside or a precursor thereof, wherein an expression level of a transcription factor of a phospholipid biosynthetic gene is changed, as compared with its intrinsic expression level, 
     
     
         2 . The recombinant yeast of  claim 1 , wherein the transcription factor of the phospholipid biosynthetic gene is one or more selected from the group consisting of INO2(INOsitol requiring 2), INO4(INOsitol requiring 4), and OPI1(OverProducer of Inositol 1). 
     
     
         3 . The recombinant yeast of  claim 2 , wherein an expression level of INO2 (INOsitol requiring 2) or INO4 (INOsitol requiring 4) or expression levels of both of them is/are increased, as compared with their intrinsic expression levels, or an expression level of OPI1 (OverProducer of Inositol 1) is decreased, as compared with its intrinsic expression level. 
     
     
         4 . The recombinant yeast of  claim 3 , wherein the INO2 (INOsitol requiring 2) gene is composed of a nucleotide sequence of SEQ ID NO: 1, the INO4 (INOsitol requiring 4) gene is composed of a nucleotide sequence of SEQ ID NO: 2, and the OPI1 (OverProducer of Inositol 1) gene is composed of a nucleotide sequence of SEQ ID NO: 3, 
     
     
         5 . The recombinant yeast of  claim 1 , wherein the yeast is selected from the group consisting of  Saccharomyces cerevisiae  ( S. cerevisiae ), Saccharomyces bayanus  ( S. bayanus ),  Saccharomyces boulardii  ( S. boulardii ),  Saccharomyces bulderi  ( S. bulderi ),  Saccharomyces cariocanus  ( S. cariocanus ),  Saccharomyces cariocus  ( S. cariocus ),  Saccharomyces chevalieri  ( S. chevalieri ),  Saccharomyces dairenensis  ( S. dairenensis ),  Saccharomyces ellipsoideus  ( S. ellipsoideus ),  Saccharomyces eubayanus  ( S. eubayanus ),  Saccharomyces exiguus  ( S. exiguus ),  Saccharomyces florentinus  ( S. florentinus ),  Saccharomyces kluyveri  ( S. kluyveri ),  Saccharomyces martiniae  ( S. martiniae ),  Saccharomyces monacensis  ( S. monacensis ),  Saccharomyces norbensis  ( S. norbensis ),  Saccharomyces paradoxus  ( S. paradoxus ),  Saccharomyces pastorianus  ( S. pastorianus ),  Saccharomyces spencerorum  ( S. spencerorum ),  Saccharomyces turicensis  ( S. turicensis ),  Saccharomyces unisporus  ( S. unisporus ),  Saccharomyces uvarum  ( S. uvarum ), and  Saccharomyces zonatus  ( S. zonatus ) 
     
     
         6 . The recombinant yeast of  claim 1 , wherein an expression level of a gene involved in ginsenoside synthesis is increased, as compared with its intrinsic expression level. 
     
     
         7 . The recombinant yeast of  claim 6 , wherein the gene is one or more genes selected from the group consisting of PgDDS (Panax ginseng, dammarenediol-II synthase), PgPPDS (Panax ginseng cytochrome P450 CYP716A47), PgCPR (Panax ginseng NADPH-cytochrome P450 reductase), tHMG1 ( S. cerevisiae  HMG-CoA reductase), and PgSE (Panax ginseng, squalene epoxidase). 
     
     
         8 . The recombinant yeast of  claim 1 , wherein the precursor is squalene or 2,3-oxidosqualene. 
     
     
         9 . A method of preparing a recombinant yeast having improved ability to produce ginsenoside, the method comprising the step of changing an expression level of a transcription factor of a phospholipid biosynthetic gene in a ginsenoside-producing yeast strain. 
     
     
         10 . The method of  claim 9 , wherein an expression level of the phospholipid biosynthetic gene, INO2(INOsitol requiring 2) or INO4 (INOsitol requiring 4), or expression levels of both of them is/are increased, as compared with their intrinsic expression levels, or an expression level of OPI1(OverProducer of Inositol 1) is decreased, as compared with its intrinsic expression level, 
     
     
         11 . The method of  claim 9 , wherein the ginsenoside-producing yeast strain has increased expression levels of one or more genes selected from the group consisting of PgDDS (Panax ginseng, dammarenediol-II synthase), PgPPDS (Panax ginseng cytochrome P450 CYP716A47), PgCPR (Panax ginseng NADPH-cytochrome P450 reductase), tHMG1 ( S. cerevisiae  HMG-CoA reductase), and PgSE (Panax ginseng, squalene epoxidase), as compared with their intrinsic expression levels. 
     
     
         12 . A method of preparing a recombinant yeast having improved ability to produce a ginsenoside precursor, the method comprising the step of changing an expression level of a transcription factor of a phospholipid biosynthetic gene in a ginsenoside precursor-producing yeast strain. 
     
     
         13 . The method of preparing the recombinant yeast of  claim 12 , wherein the ginsenoside precursor includes squalene and 2,3-oxidosqualene. 
     
     
         14 . The method of preparing the recombinant yeast of  claim 12 , wherein an expression level of the phospholipid biosynthetic gene, INO2 (INOsitol requiring 2) or INO4 (INOsitol requiring 4), or expression levels of both of them is/are increased, as compared with their intrinsic expression levels, or an expression level of OPI1 (OverProducer of Inositol 1) is decreased, as compared with its intrinsic expression level. 
     
     
         15 . A method of preparing ginsenoside or a precursor thereof, the method comprising the step of culturing the recombinant yeast of  claim 1 .