# US 8,541,211 B2 · Generation of triacylglycerols > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 8,541,211 B2 - Document ID: US8541211B2 - Status: Granted - Kind: B2 - Application number: 13/545,852 - Filed: 2012-07-10 - Published: 2013-01-10 - Granted: 2013-09-24 - Earliest claimed: 2008-01-07 - Assignees: Bunge Oils, Inc. - Applicants: Christopher L. G. Dayton - Inventors: Christopher L. G. Dayton - Web page: https://patentgrep.com/patent/US8541211B2 ## CPC - C11B 13/00: Recovery of fats, fatty oils or fatty acids from waste materials - C11C 1/045: {using enzymes or microorganisms, living or dead} - C11C 3/08: with fatty acids - C12Y 301/01004: Phospholipase A2 (3.1.1.4) - C12Y 301/01032: Phospholipase A1 (3.1.1.32) - C12Y 301/04003: Phospholipase C (3.1.4.3) - C12Y 301/04011: Phosphoinositide phospholipase C (3.1.4.11) - Y02W 30/74: Recovery of fats, fatty oils, fatty acids or other fatty substances, e.g. lanolin or waxes - C11B: PRODUCING, e.g. BY PRESSING RAW MATERIALS OR BY EXTRACTION FROM WASTE MATERIALS, REFINING OR PRESERVING FATS, FATTY SUBSTANCES, e.g. LANOLIN, FATTY OILS OR WAXES; ESSENTIAL OILS; PERFUMES (drying-oils C09F) - C11C: FATTY ACIDS FROM FATS, OILS OR WAXES; CANDLES; FATS, OILS OR FATTY ACIDS BY CHEMICAL MODIFICATION OF FATS, OILS, OR FATTY ACIDS OBTAINED THEREFROM - C12Y: ENZYMES - Y02W: CLIMATE CHANGE MITIGATION TECHNOLOGIES RELATED TO WASTEWATER TREATMENT OR WASTE MANAGEMENT ## Abstract A method is disclosed for the generation of triacylglycerols from gums that have been separated from an oil product. The gums are treated with an enzyme having PLC activity, which results in the formation of diacylglycerols and phosphates, and treated with an enzyme having PLA activity, which results in the formation of lyso-phospholipids and free fatty acids. The diacylglycerols and the free fatty acids from these two separate reactions then combine in the presence of the enzymes to generate new triacylglycerol molecules. ## Claims ### Claim 1 A method of generating triacylglycerols comprising (a) providing an oil composition containing a quantity of oil gums, wherein the gums comprise phospholipids, (b) treating the oil composition with one or more enzymes having PLA activity to generate free fatty acids, (c) treating the oil composition with one or more enzymes having PLC activity to generate diacylglycerols, (d) reacting the fatty acids with the diacylglycerols in the presence of at least one of the enzymes having PLA or PLC activity, thereby forming triacylglycerols. ### Claim 2 (depends on 1) The method of claim 1, wherein steps (b) and (c) occur substantially simultaneously. ### Claim 3 (depends on 1) The method of claim 1, wherein step (b) occurs before step (c). ### Claim 4 (depends on 1) The method of claim 1, wherein step (c) occurs before step (b). ### Claim 5 (depends on 1) The method of claim 1, wherein the total duration of the treating of the enzymes with the oil composition in steps (b) and (c) and the reacting in step (d) is no more than about four hours. ### Claim 6 (depends on 5) The method of claim 5, wherein the total duration of the treating of the enzymes with the oil composition in steps (b) and (c) and the reacting in step (d) is about thirty minutes. ### Claim 7 (depends on 1) The method of claim 1, wherein the one or more enzymes having PLA activity are selected from the group consisting of a phospholipase A1 enzyme and a phospholipase A2 enzyme. ### Claim 8 (depends on 1) The method of claim 1, wherein the one or more enzymes having PLC activity are selected from the group consisting of a phospholipase C enzyme and a phosphatidyl-inositol specific phospholipase C enzyme. ### Claim 9 (depends on 1) The method of claim 1, wherein the treating of steps (b) or (c) or the reacting of step (d) comprises treating or reacting at a pH of about 8 or less. ### Claim 10 (depends on 9) The method of claim 9, wherein the pH is about 3-7. ### Claim 11 (depends on 1) The method of claim 1, wherein the treating of the enzymes with the oil composition in steps (b) and (c) and the reacting in step (d) occurs at a temperature of about 40-80° C. ### Claim 12 (depends on 11) The method of claim 11, wherein the treating of the enzymes with the oil composition in steps (b) and (c) and the reacting in step (d) occurs at a temperature of about 40-60° C. ### Claim 13 (depends on 1) The method of claim 1, wherein the oil composition comprises a crude oil. ### Claim 14 (depends on 1) The method of claim 1, wherein in steps (c) or (d), the PLC enzyme is present in a quantity of about 30 ppm of active enzyme or less. ### Claim 15 (depends on 14) The method of claim 14, wherein the PLC enzyme is present in a quantity of about 20 ppm of active enzyme or less. ### Claim 16 (depends on 15) The method of claim 15, wherein the PLC enzyme is present in a quantity of about 10 ppm of active enzyme or less. ### Claim 17 (depends on 1) The method of claim 1, wherein in steps (b) or (d), the PLA enzyme is present in a quantity of about 2 ppm of active enzyme or less. ### Claim 18 (depends on 17) The method of claim 17, wherein the PLA enzyme is present in a quantity of about 1 ppm of active enzyme or less. ### Claim 19 (depends on 18) The method of claim 18, wherein the PLA enzyme is present in a quantity of about 0.5 ppm of active enzyme or less. ### Claim 20 (depends on 1) The method of claim 1, wherein the treating of steps (b) or (c) comprise shear mixing the oil composition and the respective PLA or PLC enzymes to obtain a mixture. ### Claim 21 (depends on 20) The method of claim 20, wherein the shear mixing continues for a duration of at least 45 seconds. ### Claim 22 (depends on 20) The method of claim 20, wherein the treating of steps (b) or (c) further comprises adding a quantity of water to the mixture. ### Claim 23 (depends on 22) The method of claim 22, wherein the quantity of water is at least about 1.5% by weight of the mixture. ### Claim 24 (depends on 23) The method of claim 23, wherein the quantity of water is at least about 3.0% by weight of the mixture. ### Claim 25 (depends on 24) The method of claim 24, wherein the quantity of water is at least about 4.5% by weight of the mixture.