![]() ![]() The method of claim 1, wherein the method of heterotrophically culturing Euglena gracilis is continuous culturing.ġ5. ![]() ![]() The method of claim 1, wherein the Euglena gracilis is grown in batch mode until the final wet cell weight is between 5 to 250 g/L (1.6 to 80 g/L dry cell weight).ġ4. The method of claim 10, wherein the vitamin is selected from the group consisting of biotin (vitamin B7), thiamine (vitamin Bl), riboflavin (vitamin B2), niacin (vitamin B3), pantothenic acid (vitamin B5), Pyridoxine (vitamin B6), Cyanocobalamin (vitamin B12), vitamin C, vitamin D, folic acid, vitamin A, vitamin B12, vitamin E, vitamin K, andġ3. The method of claim 10, wherein the metal is selected from the group consisting of iron (III) chloride, manganese chloride, zinc sulfate, sodium molybdate, zinc chloride, boric acid, copper chloride, ammonium heptamolybdate, and combinations thereof.ġ2. The method of claim 1, wherein the culture media further comprises a metal, a vitamin, and combinations thereof.ġ1. The method of claim 1, wherein the one or more salt selected from the group consisting of ammonium nitrate, sodium nitrate, monopotassium phosphate, magnesium sulfate, magnesium sulfate heptahydrate, calcium chloride, calcium chloride dihydrate, calcium sulfate, and combinations thereof.ġ0. The method of claim 1, wherein the one or more nitrogen source is selected from the group consisting of yeast extract, ammonium sulfate, glycine, urea, alanine, asparagine, corn steep, liver extract, lab lemco, peptone, skimmed milk, soy milk, tryptone, beef extract, tricine, plant source peptone, pea protein, brown rice protein, soybean peptone, potato liquor, and combinations thereof.ĩ. The method of claim 1, wherein the one or more alcohol is selected from the group consisting of ethanol, methanol, isopropanol, and combinations thereof.Ĩ. The method of claim 1, wherein the one or more sugar is selected from the group consisting of glucose, fructose, galactose, lactose, maltose, sucrose, molasses, glycerol, xylose, dextrose, honey, com syrup, and combinations thereof.ħ. ![]() The method of claim 1, wherein the one or more oil is selected from the group consisting of vegetable oil, soybean oil, coconut oil, olive oil, peanut oil, fish oil, avocado oil, palm oil, flax oil, corn oil, cottonseed oil, canola oil, rapeseed oil, sunflower oil, sesame oil, grapeseed oil, safflower oil, rice bran oil, propionate, palm kernel oil, cuphea oil, camelina sativa oil, mustard seed oil, cashew nut oil, oat oil, lupine oil, kenaf oil, calendula oil, hemp oil, coffee oil, linseed oil, hazelnut oil, euphorbia oil, pumpkin seed oil, coriander oil, camellia oil, rice oil, tung oil tree oil, cocoa oil, copra oil, opium poppy oil, castor bean oil, pecan oil, jojoba oil, jatropha oil, macadamia oil, Brazil nut oil, and combinations thereof.Ħ. The method of claim 3, wherein the carboxylic acid is citric acid, citrate, fumaric acid, fumarate, malic acid, malate, pyruvic acid, pyruvate, succinic acid, succinate, acetic acid, acetate, lactic acid, lactate, or combinations thereof.ĥ. The method of claim 1, wherein the one or more carbon source is selected from an oil, a sugar, an alcohol, carboxylic acids, ferulic acid, and combinations thereof.Ĥ. The method of claim 1, wherein the three cultivation stages include first stage cultivation, second stage cultivation, and third stage cultivation.ģ. Wherein the culturing occurs in three cultivation stages.Ģ. Maintaining an environment with substantially no light Maintaining a temperature of about 20☌ to about 30☌ and Maintaining a pH of between about 2.0 to about 4.0 A method of heterotrophically culturing a Euglena gracilis comprising:Ĭulturing the Euglena gracilis in a culture media containing one or more carbon source, one or more nitrogen source, one or more sugar, one or more alcohol, one or more oil, and one or more salt ![]()
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