Tailored oils

Abstract

Recombinant DNA techniques are used to produce oleaginous recombinant cells that produce triglyceride oils having desired fatty acid profiles and regiospecific or stereospecific profiles. Genes manipulated include those encoding stearoyl-ACP desaturase, delta 12 fatty acid desaturase, acyl-ACP thioesterase, ketoacyl-ACP synthase, and lysophosphatidic acid acyltransferase. The oil produced can have enhanced oxidative or thermal stability, or can be useful as a frying oil, shortening, roll-in shortening, tempering fat, cocoa butter replacement, as a lubricant, or as a feedstock for various chemical processes. The fatty acid profile can be enriched in midchain profiles or the oil can be enriched in triglycerides of the saturated-unsaturated-saturated type.

1 .- 84 . (canceled) 
     
     
         85 . A recombinant cell of the genus  Prototheca  or  Chlorella,  said recombinant cell comprising a knockout or knockdown of an endogenous Fatty acyl-ACP thioesterase gene, and further comprising exogenous nucleic acids that encode lysophosphatidic acid acyltransferase (LPAAT). 
     
     
         86 . The recombinant cell of  claim 85 , wherein said exogenous nucleic acids that encode LPAAT encodes an LPPAT having at least 90% amino acid sequence identity to an LPAAT selected from the group consisting of SEQ ID NOs: 16, 77, 78, 79 and 157. 
     
     
         87 . The recombinant cell of  claim 86 , wherein said exogenous nucleic acids that encode LPAAT encodes an LPPAT having at least 95% amino acid sequence identity to an LPAAT selected from the group consisting of SEQ ID NOs: 16, 77, 78, 79 and 157. 
     
     
         88 . The recombinant cell of claim of  claim 86 , wherein said exogenous nucleic acids encodes an LPAAT comprising the amino acid sequences of SEQ ID NO: 157. 
     
     
         89 . The recombinant cell of claim of  claim 86 , wherein said exogenous nucleic acids encodes an LPAAT comprising the amino acid sequences of SEQ ID NOs: 17, 77, 78 or 79. 
     
     
         90 . The recombinant cell of  claim 89 , wherein said exogenous nucleic acids that encode LPAAT are nucleic acids having at least 90% sequence identity to the nucleic acids selected from the group consisting of SEQ ID NOs: 18, 80, 81, 82, 83, 84 and 85. 
     
     
         91 . The recombinant cell of  claim 85 , wherein said exogenous nucleic acids that encode LPAAT are nucleic acids having at least 95% sequence identity to the nucleic acids selected from the group consisting of SEQ ID NOs: 18, 80, 81, 82, 83, 84 and 85. 
     
     
         92 . The recombinant cell of  claim 85 , wherein triglyceride oil produced by the recombinant cell is enriched in stearate-oleate-stearate (SOS) triglycerides. 
     
     
         93 . The recombinant cell of  claim 92 , wherein the SOS triglyceride content comprises at least 50% of the triglyceride oils. 
     
     
         94 . The recombinant cell of  claim 93 , wherein the SOS triglyceride content comprises at least 60% of the triglyceride oils. 
     
     
         95 . The recombinant cell of  claim 85 , wherein triglyceride oil produced by the recombinant cell is enriched in mid-chain fatty acid. 
     
     
         96 . The recombinant cell of  claim 95 , wherein the triglyceride oil is enriched in C12:0. 
     
     
         97 . The method of  claim 85 , wherein said recombinant cell is of the genus  Prototheca.    
     
     
         98 . The method of  claim 97 , wherein said recombinant cell is a  Prototheca moriformis  cell. 
     
     
         99 . The method of  claim 92 , wherein said recombinant cell is of the genus  Prototheca.    
     
     
         100 . The method of  claim 99 , wherein said recombinant cell is a  Prototheca moriformis  cell. 
     
     
         101 . A method of producing oil, the method comprising the steps of:
 a. cultivating a recombinant cell of the genus  Prototheca  or  Chlorella,  said recombinant cell comprising a knockout or knockdown of an endogenous Fatty acyl-ACP thioesterase gene, and further comprising exogenous nucleic acids that encode lysophosphatidic acid acyltransferase (LPAAT); and   b. isolating the oil from said recombinant cell.   
     
     
         102 . The method of  claim 101 , wherein said recombinant cell is of the genus  Prototheca.    
     
     
         103 . The method of  claim 102 , wherein said recombinant cell is a  Prototheca moriformis  cell. 
     
     
         104 . An oil produced by the steps of:
 a. cultivating a recombinant cell of the genus  Prototheca  or  Chlorella,  said recombinant cell comprising a knockout or knockdown of an endogenous Fatty acyl-ACP thioesterase gene, and further comprising exogenous nucleic acids that encode lysophosphatidic acid acyltransferase (LPAAT); and   b. isolating the oil from said recombinant cell.