Food Science and Human Nutrition Patents Food Science and Human Nutrition 9-9-2010 Enzyme-Assisted De-Emulsification of Aqueous Lipid Extracts Christopher Penet Peter Birschbach Buddhi P. Lamsal [email protected] Lawrence A. Johnson Iowa State University, [email protected] Stephanie E. Glatz Iowa State University See next page for additional authors Follow this and additional works at: http://lib.dr.iastate.edu/fshn_hs_patents Part of the Food Chemistry Commons, and the Food Processing Commons Recommended Citation Penet, Christopher; Birschbach, Peter; Lamsal, Buddhi P.; Johnson, Lawrence A.; Glatz, Stephanie E.; Glatz, Charles E.; Zhang, Cheng; and Wu, Jianping, "Enzyme-Assisted De-Emulsification of Aqueous Lipid Extracts" (2010). Food Science and Human Nutrition Patents. 2. http://lib.dr.iastate.edu/fshn_hs_patents/2 This Patent Application is brought to you for free and open access by the Food Science and Human Nutrition at Iowa State University Digital Repository. It has been accepted for inclusion in Food Science and Human Nutrition Patents by an authorized administrator of Iowa State University Digital Repository. For more information, please contact [email protected]. Enzyme-Assisted De-Emulsification of Aqueous Lipid Extracts Abstract Compositions and processes for destabilizing an oil-in-water emulsion resulting from the aqueous solvent extraction of plant oils are disclosed. The processes comprise the use of one or more enzyme activities including phospholipase and protease activity. The processes are useful for improving the extraction of oil from oilseeds, as well as for obtaining more desirable proteins from those oilseeds. Keywords Chemical and Biological Engineering Disciplines Food Chemistry | Food Processing | Food Science Authors Christopher Penet, Peter Birschbach, Buddhi P. Lamsal, Lawrence A. Johnson, Stephanie E. Glatz, Charles E. Glatz, Cheng Zhang, and Jianping Wu This patent application is available at Iowa State University Digital Repository: http://lib.dr.iastate.edu/fshn_hs_patents/2 US 20100227042A1 (19) United States (12) Patent Application Publication (10) Pub. No.: US 2010/0227042 A1 Penet et a]. (43) Pub. Date: Sep. 9, 2010 (54) ENZYME-ASSISTED DE-EMULSIFICATION (86) PCT No.: PCT/US2007/024897 OF AQUEOUS LIPID EXTRACTS § 371 (0X1), (2), (4) Date: Apr. 14, 2010 (76) Inventors: Christopher Penet, Manakin R l t d U s A l. t. D t Sabot, VA (US); Peter Birschbach, e a e l ' PP lea Ion a a Rochester, NY (US); Buddhi P, (60) Provisional application No. 60/876,879, ?led on Dec. Lamsal, Manhattan, KS (US); 22, 2006 Lawl‘ellce A- Johllsoll, Ames, IA Publication Classi?cation (US); Stephanie E. Glatz, Ames, _ (51) Int. Cl. IA (US), Charles E. Glatz, Ames, A23L 1/28 (200601) IA (US), Cheng Zhang, Ames, IA _ . B01D 17/04 (2006.01) (US), Jlanplng Wu, Edmonton CUB 1/00 (200601) (CA) A23L 1/305 (2006.01) C07K 2/00 (2006.01) Correspondence Address; (52) U.S. Cl. ....... .. 426/655; 435/266; 554/115; 530/350; DANISCO US INC. 426/656 ATTENTION: LEGAL DEPARTMENT (57) ABSTRACT 925 PAGE MILL ROAD PALO ALTO, CA 94304 (US) Compositions and processes for destabilizing an oil-in-Water emulsion resulting from the aqueous solvent extraction of plant oils are disclosed. The processes comprise the use of one (21) App1.No.: 12/520,028 or more enzyme activities including phospholipase and pro tease activity. The processes are useful for improving the extraction of oil from oilseeds, as Well as for obtaining more (22) PCT Filed: Dec. 4, 2007 desirable proteins from those oilseeds. Patent Application Publication Sep. 9, 2010 Sheet 1 0f 4 US 2010/0227042 A1 LysoMaxTM? ' ‘i . FIG. 1 Patent Application Publication Sep. 9, 2010 Sheet 2 0f 4 US 2010/0227042 A1 2.0M FIG. 2 Patent Application Publication Sep. 9, 2010 Sheet 3 0f 4 US 2010/0227042 A1 Soybeans i Cracking i Dehulling i Conditioning ‘ 4—- Water Flaking i Extrusion Enzyme-assisted Aqueous Extraction l Centrifugation —-—> Solids i Aqueous Extract Skim Fraction 4___ Settling ———> Free Oil 4' Cream 4, ‘ Enzyme-assistedemulsi?cation De ——‘, _ _ Free Oil Sk1m Fraction FIG. 3 Patent Application Publication Sep. 9, 2010 Sheet 4 0f 4 US 2010/0227042 A1 El Control E Phospholipase C 120 E1 Genencor enzyme cocktail E Freeze-thaw 109 El Heat, 95C 100 8D Q\° 60 40 20 1212131112 0 I lnsolubles Residual Skim Free Oil Total Cream FIG. 4 US 2010/0227042 A1 Sep. 9, 2010 ENZYME-ASSISTED DE-EMULSIFICATION [0007] There is a need therefore in the art for processing OF AQUEOUS LIPID EXTRACTS steps to improve aqueous extraction methods for obtaining oils from plant sources, especially oilseeds. There is also a FIELD OF THE INVENTION need to increase the e?iciency or yields of such processes. [0001] This pertains to the use of enzymes to aid in the SUMMARY extraction of lipids from plants or plant tissues. More particu larly, it pertains to compositions comprising enZymes for [0008] In several of its various aspects, processes are pro de-emulsifying emulsions obtained from aqueous extraction vided for using de-emulsifying oil-in-Water emulsions of lipids from plants, and improved methods for their use. obtained from aqueous extraction of plant tissues, particu larly oilseeds, for example by the use of various enZymes. BACKGROUND Also provided are methods for obtaining oil from oilseeds Wherein the method includes aqueous extraction and enZy [0002] Plant-derived lipids, particularly oils, are a major matic de-emulsi?cation of a resulting oil-in-Water emulsion. source of lipids for food processing, and for industrial feed Oil derived by the processes and methods, as Well as compo stock. More recently they are of interest and use as alterna sitions comprising the oil such as food, consumer products tives to petrochemicals for fuels. Plant lipids can be derived and industrial feedstocks are also provided. Compositions from one or more parts of a plant, shrub, or tree. In various comprising enZymes capable of de-emulsifying an oil-in plants, the root, stem, bark, leaves, ?oWers, seeds, fruits, or Water emulsion and further comprising an oil-in-Water emul other parts may serve as a source of oil. Such lipids can be sion are also provided herein. extracted mechanically, eg through the application of exter [0009] In one aspect, provided are processes for destabiliZ nal pressure, or chemically, eg through organic or aqueous ing an emulsion comprising an oil phase and an aqueous solvent extraction processes, or combination processes. phase, Where the emulsion has been produced in an aqueous [0003] Plant oils are often classi?ed as either essential or solvent extraction of a lipid from an oilseed. The process ?xed. Essential oils are volatile oils typically derived from comprises the step of contacting the emulsion With at least tissues other than the seed of a plant. Fixed oils, also called one enZyme activity including at least a phospholipase or a “fatty oils” include oils derived from plants sources, for protease, or a combination thereof, under conditions alloWing example, soybean, corn, canola (rapeseed), sun?ower, saf activity of at least the phospholipase or the protease, or the ?oWer, peanut, coconut, copra, palm, cottonseed, olive, combination, for a time su?icient to destabiliZe the emulsion. sesame, ?axseed, and others. In certain preferred embodiments the Water phase is the con [0004] Oils obtained from oilseed plants are knoWn for tinuous phase of the emulsion and the oil phase is the discon their uses in food, and food products, as Well as for soaps, tinuous phase of the emulsion. detergents, lotions, lubricants, insecticides, paints, coatings, [0010] In another aspect, processes are provided for obtain inks, and other industrial or consumer products. Although the ing oil from an oilseed. The processes generally comprise the vast majority of oilseeds oils are extracted With an organic steps of: solvent extraction process, some are noW extracted through [0011] (a) providing an oil-containing oilseed fraction; aqueous extraction. [0012] (b) contacting the oil-containing oilseed fraction [0005] Extraction With organic solvents is faster and more With an aqueous extractant to form an extracted oilseed e?icient, providing higher yields of oil. This is by far the fraction; dominant commercial process for extraction of, for example, [0013] (c) separating the extracted oilseed fraction into soybean oil, With greater than 90% of all soy bean oil an aqueous phase, an oil-in-Water emulsion, and an extracted With hexane or other organic solvents. HoWever, insoluble phase; such solvents pose substantial safety issues, including the risk [0014] (d) contacting the oil-in-Water emulsion With at of ?re and explosions from solvents, as Well as exposure risks least one enZyme activity under conditions permitting to personnel. Solvent extraction processes require expensive enZyme activity for a time suf?cient to destabiliZe the plants that are equipped to handle the solvents, as Well as the emulsion; and required the safety measures. In addition, the large-scale use [0015] (e) separating the destabiliZed emulsion into an of organic solvents raises Waste management concerns. Fur aqueous phase, an oil phase, and an insoluble phase. thermore, for applications in food processing, the solvent [0016] The processes result in obtaining oil from the oil based extractions may result in denatured, altered, or less seed. In one embodiment, the oil-containing oilseed fraction functional proteins in the remaining, extracted portion of the comprises cells and the process further comprises the step of oilseed (eg the “meal”), decreasing the value or use of this disrupting the cells prior to contacting the oilseed fraction potentially more valuable portion of the oilseed. With the aqueous extractant. [0006] Aqueous processes for extracting oils from oilseeds [0017] Also provided herein are plant-derived oils prepared have been developed in recent years. See Freitas et al., Fell/ by the above disclosed processes, as Well as a host of food Lipid 99: 333-337, 1997; and Caetano et al., La Rivisla Ital products comprising the oil so obtained.