Anaplerotic Oil Production In Microbials

Abstract

Disclosed are techniques and systems for producing microbials having anaplerotic oils that are rich in odd-chain fatty acids, and other beneficial components, at higher concentrations than those present in other natural dietary sources of OCFA, at lower cost, and higher production yield. Such compositions can comprise pentadecanoic and heptadecanoic fatty acids. The techniques described herein include methods for producing and deriving such compositions rich in odd-chain fatty acids from microbials, including microalgae and yeasts/fungi.

What is claimed is: 
     
         1 . A method for producing anaplerotic oil from microorganisms, the method comprising:
 adding microorganisms to a culture medium;   adding a carbon source to the culture medium, wherein the carbon source is taken up and used by the microorganisms to support growth of the microbial mass;   after adding the carbon source to the culture medium, adding propionate to the culture medium, wherein the propionate is taken up and used by the microorganisms for production of odd-chain fatty acids (OCFAs), resulting in cultured microorganisms with enhanced synthesis of saturated tridecanoic (C13:0), pentadecanoic (C15:0), and heptadecanoic (C17:0) OFCAs; and   harvesting an anaplerotic oil from the cultured microorganisms.   
     
     
         2 . The method of  claim 1 , wherein:
 at least five percent of the total fatty acids (TFAs) of the anaplerotic oil are saturated tridecanoic (C13:0), pentadecanoic (C15:0), and heptadecanoic (C17:0) OCFAs; and   the saturated tridecanoic (C13:0), pentadecanoic (C15:0), and heptadecanoic (C17:0) OCFAs make up at least one percent of the cell dry weight (CDW) of the microbial mass.   
     
     
         3 . The method of  claim 1 , wherein adding propionate comprises adding the propionate to produce OCFAs in a range of 5 and 70% TFAs. 
     
     
         4 . The method of  claim 1 , wherein the propionate and the carbon source are added at a weight-to-weight ratio below 0.1 of propionate to carbon source. 
     
     
         5 . The method of  claim 1 , wherein adding propionate comprises adding the propionate in a batch system into the culture medium. 
     
     
         6 . The method of  claim 1 , wherein adding propionate comprises adding the propionate in a fed-batch system into the culture medium. 
     
     
         7 . The method of  claim 6 , wherein the propionate is fed at a rate of greater than zero and up to 3 grams per liter per day. 
     
     
         8 . The method of  claim 1 , wherein the propionate is added on demand using a pH-auxostat fed-batch system to maintain a desired pH of the culture medium. 
     
     
         9 . The method of  claim 1 , wherein the propionate is added when the culture medium is at a pH above 5 to reduce propionic acid toxicity. 
     
     
         10 . The method of  claim 1 , wherein the propionate is added at one or more of:
 following a rise in the pH of the culture medium;   the lipid phase of the microorganisms in the culture medium to mitigate exposure to propionate toxicity of the microorganisms; and   the end of the protein phase and start of lipogenesis of the microorganisms.   
     
     
         11 . The method of  claim 1 , wherein the microorganisms are oleaginous and can produce lipids to at least 20% dry cell weight. 
     
     
         12 . The method of  claim 1 , wherein cyanocobalamin is added to or subtracted from the culture medium to modify propionic acid deposition in OCFA. 
     
     
         13 . The method of  claim 12 , wherein the concentration of the cyanocobalamin in the culture medium is below 0.4 μM. 
     
     
         14 . The method of  claim 1 , wherein at least 0.05 gram of propionate is added per 1 gram of biomass produced. 
     
     
         15 . The method of  claim 1 , wherein the microorganisms are cultured under aerobic conditions by continuously supplying oxygen to the culture. 
     
     
         16 . The method of  claim 15 , wherein the oxygen in the culture is maintained at 10% saturation or greater. 
     
     
         17 . The method of  claim 1 , wherein the propionate is in the form of propionic acid, an anion of propionic acid, a salt of propionic acid, and/or an ester of propionic acid. 
     
     
         18 . The method of  claim 1 , wherein the carbon source is one or more of: arabinose, fructose, galactose, glucose, glycerol, lactose, maltose, mannose, molasses, ribose, and sucrose. 
     
     
         19 . The method of  claim 1 , wherein the carbon source is one or more of: acetate, acetic acid, and citric acid. 
     
     
         20 . The method of  claim 1 , wherein the carbon source is one or more of: glycine, peptone, plant-based hydrolysate, partial or complete hydrolysates of starch, and yeast extract.