Oil degumming methods
In alternative embodiments, the invention provides phosphatidylinositol-specific phospholipase C (PI-PLC) enzymes, nucleic acids encoding them, antibodies that bind specifically to them, and methods for making and using them. Industrial methods and products comprising use of these phospholipases are also provided. In certain embodiments, provided herein are methods for hydration of non hydratable phospholipids (NHPs) within a lipid matrix. The methods enable migration of NHPs to an oil-water interface thereby allowing the NHPs to be reacted and/or removed from the lipids. In certain embodiments, provided is a method for removing NHPs, hydratable phospholipids, and lecithins from vegetable oils to produce a degummed oil or fat product that can be used for food production and/or non-food applications. In certain embodiments, provided herein are methods for hydration of NHPs followed by enzymatic treatment and removal of various phospholipids and lecithins. The methods provided herein can be practiced on either crude or water-degummed oils.
1. A method for degumming a crude oil comprising:
i-a) mixing an aqueous acid with the crude oil to obtain an acidic mixture having pH of less than about 4;
i-b) mixing a base with the acidic mixture to obtain a reacted mixture having pH of about 6-9, wherein the mixing in steps i-a) and/or i-b) creates an emulsion that comprises at least about 60% of an aqueous phase by volume in droplet size between about 15 μm to about 45 μm in size; and
i-c) mixing a phospholipase enzyme with the emulsion in step (i-b), wherein the phospholipase enzyme comprises SEQ ID NO:8.
2. The method of claim 1 , wherein the emulsion comprises at least about 90% of the aqueous phase by volume in droplet size of about 15-40 μm in size.
3. The method of claim 1 , wherein the acid is selected from the group consisting of phosphoric acid, acetic acid, citric acid, tartaric acid, succinic acid, and a mixture thereof.
4. The method of claim 1 , wherein the pH of the acidic mixture in step i-a) is about 1 to about 4.
5. A method for degumming a crude oil comprising:
i-a) mixing an aqueous acid with the crude oil to obtain an acidic mixture having pH of less than about 4;
i-b) mixing a base with the acidic mixture to obtain a reacted mixture having pH of about 6-9, wherein the mixing in steps i-a) and/or i-b) creates an emulsion that comprises an aqueous phase in average droplet size between about 15-35 μm; and
i-c) mixing a phospholipase enzyme with the emulsion in step (i-b), wherein the phospholipase enzyme comprises SEQ ID NO:8.
6. The method of claim 1 , wherein the enzyme is a polypeptide having SEQ ID NO: 8.
7. The method of claim 5 , wherein the enzyme is a polypeptide having SEQ ID NO: 8.