Enzymes involved in triterpene synthesis

Abstract

This invention relates to an isolated nucleic acid fragment encoding an oxidosqualene cyclase. The invention also relates to the construction of a chimeric gene encoding all or a portion of the isolated polynucleotides of the invention, in sense or antisense orientation, operably linked to a suitable regulatory sequence.

1 . An isolated polynucleotide comprising a nucleotide sequence selected from the group consisting of: 
 (a) first nucleotide sequence encoding a polypeptide of at least 100 amino acids having at least 80% identity based on the Clustal method of alignment when compared to a polypeptide selected from the group consisting of SEQ ID NOs: 2, 4, 6, and 8; and    (b) a second nucleotide sequence comprising the complement of the first nucleotide sequence.    
     
     
         2 - 42 . (canceled)  
     
     
         43 . A soybean grain produced by a transgenic plant comprising a polynucleotide selected from the group consisting of: 
 (a) a nucleotide sequence encoding a polypeptide having oxidosqualene cyclase activity, wherein the polypeptide has an amino acid sequence of at least 80% sequence identity based on the Clustal method of alignment when compared to a polypeptide of SEQ ID NO:2, 4, 6, or 8; wherein the default parameters for pairwise alignment are KTUPLE=1, GAP PENALTY=3, WINDOW=5 and DIAGONALS SAVED=5; and    (b) a full complement of the nucleotide sequence of (a).    wherein said polynucleotide is under the control of a heterologous promoter and wherein said plant has an altered level of a triterpene.    
     
     
         44 . A food product prepared from the soybean grain of  claim 43 .  
     
     
         45 . The food product of  claim 44  wherein the food product is selected from the group consisting of a soy protein product, soybean meal, soy flour, soy protein concentrate, soy milk, a dietary supplement, beans, nuts, tofu, natto, miso, and tempeh.  
     
     
         46 . A method of making a soy protein product comprising processing the grain of  claim 43 , said processing including: 
 (a) cracking said grain to remove the meats from the hulls; and    (b) flaking the meats obtained in step (a) to obtain a desired flake thickness.    
     
     
         47 . A method of making a soy product comprising inactivating an antinutrient of the grain produced by a transgenic plant comprising a polynucleotide comprising: 
 (a) a nucleotide sequence encoding a polypeptide having oxidosqualene cyclase activity, wherein the polypeptide has an amino acid sequence of at least 80% sequence identity based on the Clustal method of alignment when compared to a polypeptide of SEQ ID NO:2, 4, 6, or 8; wherein the default parameters for pairwise alignment are KTUPLE=1, GAP PENALTY=3, WINDOW=5 and DIAGONALS SAVED=5; and    (b) a full complement of the nucleotide sequence of (a)    wherein said polynucleotide is under the control of a heterologous promoter and wherein said plant has an altered level of a triterpene.    
     
     
         48 . The method of  claim 47  wherein inactivating is toasting.  
     
     
         49 . The method of  claim 47  wherein the anitnutrient is a trypsin inhibitor.  
     
     
         50 . A method of extracting bioactive compounds from the grain of  claim 43  comprising exposing the grain or portions thereof to a solvent.  
     
     
         51 . The method of  claim 50  wherein said bioactive compound is a saponin.  
     
     
         52 . A nutritional supplement produced by the method of  claim 50 .  
     
     
         53 . A pharmaceutical composition produced by the method of  claim 50.