Eicosapentaenoic acid-producing microorganisms, fatty acid compositions, and methods of making and uses thereof

Abstract

The present invention is directed to isolated microorganisms as well as strains and mutants thereof, biomasses, microbial oils, compositions, and cultures; methods of producing the microbial oils, biomasses, and mutants; and methods of using the isolated microorganisms, biomasses, and microbial oils.

1 . A method of making a biomass of a microorganism having fatty acids and a concentration of EPA, comprising:
 (a) fermenting the microorganism in a fermentor vessel to produce a biomass wherein the microorganism comprises a Thraustochytrid that produces a biomass having at least 3% EPA of the total weight of the fatty acids; and   (b) attaining a desired EPA level in the biomass by (i) adjusting the amount of the biomass in the vessel and (ii) optionally adjusting the pressure on the biomass.   
     
     
         2 . (canceled) 
     
     
         3 . A method of making a biomass of a microorganism having fatty acids and a concentration of EPA, comprising:
 (a) fermenting the microorganism in a fermentor vessel to produce a fermentation broth and a biomass wherein the microorganism comprises a Thraustochytrid that produces a biomass having at least 3% EPA of the total weight of the fatty acids; and   (b) attaining a desired EPA level in the biomass by (i) adjusting the temperature in the broth and (ii) optionally adjusting the pressure on the biomass.   
     
     
         4 . A method of making a biomass of a microorganism having fatty acids and a concentration of EPA, comprising:
 (a) fermenting the microorganism in a fermentor vessel, comprising a gas, to produce a biomass wherein the microorganism comprises a Thraustochytrid that produces a biomass having at least 3% EPA of the total weight of the fatty acids; and   (b) attaining a desired EPA level in the biomass by (i) adjusting the CO 2  levels in the gas and (ii) optionally adjusting the pressure on the biomass.   
     
     
         5 - 7 . (canceled) 
     
     
         8 . A method of increasing the concentration of EPA in a biomass of a microorganism having fatty acids and a concentration of EPA, comprising:
 (a) fermenting the microorganism in a fermentor vessel, comprising a gas, to produce a biomass wherein the microorganism comprises a Thraustochytrid that produces a biomass having at least 3% EPA of the total weight of the fatty acids; and   (b) supplementing the gas with CO 2  sufficient to increase the concentration of the EPA in the biomass.   
     
     
         9 . The method as recited in  claim 8  wherein the amount of CO 2  is greater than or equal to 2% (mole) of the gas in the chamber and the concentration of the EPA in the biomass is increased to greater than about 4%, by weight, of the total weight of the fatty acids. 
     
     
         10 - 11 . (canceled) 
     
     
         12 . The method as recited in  claim 9  wherein the concentration of EPA is increased from greater than about 4% up to about 45%, by weight, of the total weight of the fatty acids. 
     
     
         13 - 27 . (canceled) 
     
     
         28 . A biomass made according to anyone of  claims 1 ,  3 - 4 , and  8 . 
     
     
         29 . An oil extracted from a biomass of  claim 28  wherein the biomass is produced from an isolated microorganism deposited under ATCC Accession No. PTA-10208 wherein the oil comprises fatty acids wherein the fatty acids further comprise omega-3 polyunsaturated fatty acids wherein the omega-3 polyunsaturated fatty acids comprise DHA and EPA in an amount of about ≧90%, by weight, of the total amount of omega-3 polyunsaturated fatty acids and the amount of EPA, by weight, is from about 6% up to about 65% of the total amount of EPA and DHA. 
     
     
         30 - 33 . (canceled) 
     
     
         34 . An oil extracted from a biomass of  claim 28  wherein the biomass is produced from an isolated microorganism deposited under ATCC Accession No. PTA-9695 wherein the oil comprises fatty acids wherein the fatty acids further comprises omega-3 polyunsaturated fatty acids wherein the omega-3 polyunsaturated fatty acids comprise DHA and EPA in an amount of about 50 to about 70% by weight, of the total amount of omega-3 polyunsaturated fatty acids, and the amount of EPA, by weight, is from about 5% up to about 60% of the total amount of the total weight of the EPA and DHA. 
     
     
         35 . (canceled) 
     
     
         36 . A method of making a biomass of a microorganism having fatty acids and a concentration of EPA, comprising:
 (a) fermenting the microorganism in a fermentor vessel, comprising a gas, to produce a biomass wherein the microorganism comprises a Thraustochytrid that produces a biomass having at least 3% EPA of the total weight of the fatty acids wherein the microorganism comprises a Thraustochytrid that produces a biomass having at least 3% EPA of the total weight of the fatty acids; and   (b) supplementing the gas with CO 2 .   
     
     
         37 - 42 . (canceled) 
     
     
         43 . A method of making a biomass of a microorganism having increased concentration of EPA, comprising growing the microorganism in a culture medium comprising less than 0.1 mg/L vitamin B12 to produce a biomass. 
     
     
         44 - 54 . (canceled) 
     
     
         55 . A method of making a biomass of a microorganism having increased concentration of EPA, comprising growing the microorganism in a culture medium comprising less than 0.1 mg/L cobalt to produce a biomass. 
     
     
         56 - 66 . (canceled)