Production of fatty acid estolides

Patent metadata

Field	Value
Publication number	US-11512330-B2
Application number	US-202117157025-A
Country	US
Kind code	B2
Filing date	Jan 25, 2021
Priority date	Mar 26, 2015
Publication date	Nov 29, 2022
Grant date	Nov 29, 2022

Abstract

It has been found that esterification of a hydroxy-fatty acid by a lipase can be coupled with oleate hydratase (OHase) generation of that hydroxy-FA from an unsaturated FA with a cis C9-C10 double bond, e.g. oleic acid, in a single aqueous buffered reaction medium at low temperature, e.g. 30° C. A simple one-pot enzymatic method to produce fatty acid estolides from one or more triglycerides, e.g. starting from a natural plant oil, is thereby enabled in which the same lipase catalyses both the initial hydrolysis of triglyceride and the final esterification step.

First claim

The invention claimed is:

1. A method for producing one or more esters of one or more hydroxy-fatty acids comprising:
providing in a single aqueous buffered, cell-free reaction medium at least one unsaturated fatty acid with a cis C9-C10 double bond;
in a first reaction, reacting said at least one unsaturated fatty acid with a cis C9-C10 double bond in said aqueous buffered, cell-free reaction medium with an isolated cell-free oleate hydratase to produce at least one hydroxy-fatty acid; and
in a second reaction, reacting said produced at least one hydroxy-fatty acid with an isolated lipase in said aqueous buffered, cell-free reaction medium to produce one or more esters of said at least one hydroxy-fatty acid;
wherein the first reaction and the second reaction are carried out in situ at about pH 6 to 8.

2. A method as claimed in claim 1 , wherein said lipase is a lipase with non-1,3 positional specificity for hydrolysis of triglyceride.

3. A method as claimed in claim 2 wherein said lipase is selected from microbial lipases of Candida species, Pseudomonas species, Chromobacterium species, Geotrichum species, Alcaligenes species and Thermomyces species.

4. A method as claimed in claim 1 , wherein the first reaction and the second reaction are carried out at about 20-40° C.

5. A method as claimed in claim 1 wherein the first reaction and the second reaction are carried out at about pH 6.5 and about 30° C.

6. A method as claimed in claim 1 , wherein the single aqueous buffered, cell-free reaction medium is an aqueous buffered solution containing NaCl at a concentration up to about 150-200 mM.

7. A method as claimed in claim 1 , wherein said one or more esters are one or more fatty acid estolides.

8. A method as claimed in claim 7 wherein said one or more fatty acid estolides are derived from one or more mono-10-hydroxy fatty acids, said estolides being selected from a monoestolide of a hydroxy-FA with an unsaturated fatty acid, a monoestolide of a hydroxy-FA with itself or another hydroxy-FA, and higher ester oligomers formed from one or more hydroxy-FAs capped or uncapped by a non-hydroxy unsaturated fatty acid.

9. A method as claimed in claim 8 wherein 10-hydroxystearic acid is the sole hydroxy- fatty acid provided for esterification by said lipase.

10. A method as claimed in claim 1 , which further comprises extraction of one or more ester products from the reaction medium or extraction and further purification of one or more ester products and /or incorporation into a composition.

11. A method as claimed in claim 1 , wherein one or more ester products of lipase esterification are further reacted.

12. A method as claimed in claim 10 , wherein one or more ester products of lipase esterification are further reacted.

13. A method as claimed in claim 1 , wherein said isolated cell-free oleate hydratase is a recombinant oleate hydratase of Elizabethkingia meningoseptica.

14. A method as claimed in claim 1 , wherein said isolated cell-free oleate hydratase is a recombinant oleate hydratase with an N-terminal His tag covalently linked to glutaraldehyde-activated magnetic chistosan composite particles in which magnetic iron oxide particles are dispersed in a chitosan matrix.

15. A method as claimed in claim 14 , where said magnetic iron oxide particles are amino-terminated magnetic particles.

16. A method as claimed in claim 14 , wherein the first reaction is carried out at 30-35° C. and the second reaction is carried out at 60° C.

17. A method as claimed in claim 14 , wherein the first reaction and the second reaction are carried out at 50-60° C.

Assignees

• Stichting Wageningen Res
• Univ Delft Tech

Inventors

• Boeriu Carmen Gabriela
• Todea Anamaria
• Arends Isabella Wilhelmina Christina Everdina
• Otten Ludwina Gerharda

Classifications

• C12N9/88 Primary

  Lyases (4.) · CPC title

• C12P7/64

  Fats; Fatty oils; Ester-type waxes; Higher fatty acids, i.e. having at least seven carbon atoms in an unbroken chain bound to a carboxyl group; Oxidised oils or fats · CPC title

• C12N9/20

  Triglyceride splitting, e.g. by means of lipase · CPC title

• C12P7/42

  Hydroxy-carboxylic acids · CPC title

• C12Y402/01053

  Oleate hydratase (4.2.1.53) · CPC title