Compositions, Methods, and Kits for Polyunsaturated Fatty Acids from Microalgae

Abstract

The present invention provides compositions, methods, and kits comprising PUFAs produced by microalgae, in particular omega-3 and/or omega-6 fatty acids produced by members of the genus Rhodomonas , in particular Rhodomonas salina . The invention also provides compositions, methods, and kits comprising the PUFAs for the prophylactic and/or therapeutic treatment of a disease or condition, in particular a cardiovascular and/or inflammatory disease or condition.

1 .- 80 . (canceled) 
     
     
         81 . An isolated genetically modified variant Rhodomonas strain comprising at least one heterologous nucleic acid encoding an enzyme selected from the group consisting of a fatty acid synthase, a desaturase, and an elongase, wherein said strain exhibits increased polyunsaturated fatty acid levels relative to strains lacking said heterologous nucleic acids. 
     
     
         82 . The  Rhodomonas  strain of  claim 81 , wherein said nucleic acid encodes a fatty acid synthase. 
     
     
         83 . The  Rhodomonas  strain of  claim 81 , wherein said nucleic acid encodes a desaturase. 
     
     
         84 . The  Rhodomonas  strain of  claim 81 , wherein said nucleic acid encodes an elongase. 
     
     
         85 . The strain as claimed in  claim 81  to  claim 84 , which is ATCC No. PTA-9989. 
     
     
         86 . A process of making a polyunsaturated fatty acid (PUFA) composition comprising at least 8% polyunsaturated fatty acids in weight, the process comprising: extracting the polyunsaturated fatty acids from the microalgae of  claim 81 , wherein,
 (a) SDA is in an amount of about 5% to about 50% of total fatty acids;   (b) EPA is in an amount of about 2% to about 40% of total fatty acids;   (c) DHA is in an amount of about 2% to about 40% of total fatty acids; and optionally   (d) GLA is in an amount of about 0.1% to about 10% of total fatty acids; wherein the composition comprises at least 8% polyunsaturated fatty acids in weight.   
     
     
         87 . The process of  claim 86 , wherein the microalgae is a microalgae having a cell wall of reduced thickness as compared to the wild-type microalgae, wherein said cell wall of reduced thickness improves extractability and/or bioavailability of the microalgae lipid fraction. 
     
     
         88 . A process of making a composition comprising at least 5% stearidonic acid, the process comprising:
 (a) cultivating the microalgae of  claim 81  to produce a microalgae biomass; and either   (b) extracting said microalgae oil from said cultivated microalgae; or   (c) removing water from said microalgae biomass to achieve a solids content from 5 to 100% weight percent; wherein the composition comprises at least 5% stearidonic acid.   
     
     
         89 . A method for preparing a microalgae biomass composition comprising one or more PUFAs, the method comprising: culturing a microalgae of  claim 81  under a culture condition sufficient to provide a microalgae biomass comprising the one or more PUFAs, wherein the microalgae biomass is harvested after at least one logarithmic growth phase of the microalgae, wherein the omega-3 fatty acid is stearidonic acid (SDA), wherein the omega-6 fatty acid is γ-linolenic acid (GLA), wherein the SDA at harvest is at least about 5% of the total fatty acid, wherein the GLA at harvest is less than about 1% of total fatty acid. 
     
     
         90 . A biologically pure culture of the isolated  Rhodomonas  of  claim 85 . 
     
     
         91 . A method for preparing a microalgae biomass composition comprising one or more PUFAs, the method comprising: culturing a microalgae of  claim 90  under a culture condition sufficient to provide a microalgae biomass comprising the one or more PUFAs, wherein the microalgae biomass is harvested after at least one logarithmic growth phase of the microalgae, wherein the omega-3 fatty acid is stearidonic acid (SDA), wherein the omega-6 fatty acid is optionally present and is γ-linolenic acid (GLA), wherein the SDA at harvest is at least about 5% of the total fatty acid, wherein the GLA if present at harvest is less than about 1% of total fatty acid. 
     
     
         92 . A method for preparing a microalgae biomass composition comprising at least 8% polyunsaturated fatty acids in weight, the method comprising: culturing a microalgae of  claim 90  the process comprising: extracting the polyunsaturated fatty acids from the microalgae, wherein,
 (a) SDA is in an amount of about 5% to about 50% of total fatty acids; 
 (b) EPA is in an amount of about 2% to about 40% of total fatty acids; 
 (c) DHA is in an amount of about 2% to about 40% of total fatty acids; and optionally 
 (d) GLA is in an amount of about 0.1% to about 10% of total fatty acids; wherein the composition comprises at least 8% polyunsaturated fatty acids in weight. 
 
     
     
         93 . An isolated mixed culture of microalgae and bacteria, wherein said micro algae is genetically modified.