Method of Producing a Low Molecular Weight Organic Compound in a Cell

Abstract

A method of producing a low molecular weight organic compound (e.g. a plant or bacteria secondary metabolite) in increased yields involving use of a microorganism cell, which comprises a gene involved in the biosynthesis pathway leading to a low molecular weight organic aglycon compound and a glycosyltransferase gene capable of glycosylating the produced aglycon.

We claim: 
     
         1 . A method of producing a low molecular weight organic aglycon compound comprising following steps:
 a) fermenting a microorganism cell in a suitable medium where the microorganism is capable of growing, which comprises a gene encoding a product involved in the biosynthesis pathway leading to a low molecular weight organic aglycon compound and a glycosyltransferase gene encoding a glycosyltransferase capable of glycosylating the produced aglycon, under suitable conditions wherein the cell produces the aglycon and the corresponding glycosylated form of the aglycon;   b) deglycosylating the glycosylated form of the aglycon; and   c) recovering the aglycon compound;
 (i) wherein the low molecular weight organic aglycon compound has a molecular weight from 50 to 3000, and 
 (ii) wherein the glycosyltransferase is a glycosyltransferase capable of conjugating a sugar to the aglycon compound. 
   
     
     
         2 . The method of  claim 1 , wherein the microorganism cell with the glycosyltransferase during culture fermentation is capable of producing higher amounts of the glycosylated form of the aglycon as compared to the amounts of the corresponding aglycon produced by the same microorganism cell without the glycosyltransferase. 
     
     
         3 . The method of  claim 2 , wherein the microorganism cell is a yeast cell. 
     
     
         4 . The method of  claim 3 , wherein the yeast cell is a  Saccharomyces  spp. cell or a  Pichia  spp. cell. 
     
     
         5 . The method of  claim 2 , wherein the microorganism cell is a prokaryotic cell. 
     
     
         6 . The method of  claim 5 , wherein the prokaryotic cell is an  E. coli  cell. 
     
     
         7 . The method of  claim 1 , wherein the glycosyltransferase gene is a heterologous glycosyltransferase gene. 
     
     
         8 . The method of  claim 1 , wherein the glycosyltransferase is an UDPG glycosyltransferase, preferably an UDPG-glucosyltransferase. 
     
     
         9 . The method of  claim 1 , wherein the low molecular weight organic aglycon compound is an organic aglycon compound that comprises a compound that contains a Hydroxy-, Amino-, Sulfide-, or Carboxy functional group that can be glycosylated by use of the glycosyltransferase. 
     
     
         10 . The method of  claim 9 , wherein the low molecular weight organic aglycon compound is an organic aglycon compound that comprises a compound that contains Hydroxy-functional group, or an alcohol, that can be glycosylated by use of the glycosyltransferase. 
     
     
         11 . The method of  claim 9 , wherein the aglycon compound has a molecular weight from 50 to 1000 kilodaltons. 
     
     
         12 . The method of  claim 11 , wherein the aglycon compound is a secondary metabolite compound. 
     
     
         13 . The method of  claim 12 , wherein secondary metabolite compound is a plant secondary metabolite compound selected from the group consisting of Terpenoids, Alkaloids, Phenylpropanoids, Phenyl derivatives, Hexanol derivatives, Flavonoids, Coumarins, Stilbenes, Cyanohydrins, and Glucosinolates. 
     
     
         14 . The method of  claim 13 , wherein the plant secondary metabolite organic aglycon compound is vanillin. 
     
     
         15 . The method of  claim 14 , wherein the microorganism cell is a yeast cell. 
     
     
         16 . The method of  claim 15 , wherein the yeast cell is a  Saccharomyces  spp. cell or a  Pichia  spp. cell. 
     
     
         17 . The method of  claim 14 , wherein the microorganism cell is a prokaryotic cell. 
     
     
         18 . The method of  claim 17 , wherein the prokaryotic cell is an  E. coli  cell. 
     
     
         19 . The method of  claim 1 , wherein the deglycosylating step b) takes place outside the growing cell following excretion or extraction of the glycosylated form of the aglycon produced in step a) and wherein the deglycosylating is an enzymatic process mediated by a beta-glucosidase.