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Method for preparing hydroxy fatty acid

A hydroxy fatty acid, quality technology, applied in the field of enzyme engineering, can solve the problems of separation and extraction difficulties, waste of resources, etc., to achieve the effect of reducing industrial waste, realizing recycling, and solving waste of resources

Active Publication Date: 2021-08-06
SOUTH CHINA UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The by-products of oil degumming are rich in components, but separation and extraction are difficult, so they are generally discarded directly, resulting in a waste of resources

Method used

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  • Method for preparing hydroxy fatty acid
  • Method for preparing hydroxy fatty acid
  • Method for preparing hydroxy fatty acid

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] Preparation of degummed by-products. Heat the crude oil to 80°C, add phosphoric acid with 2% mass of the crude oil, and stir at high speed. When particles are generated, continue to stir and cool down to 25°C, add isothermal water, continue to stir for 10 minutes, let stand to separate layers, centrifuge, and extract the supernatant After the grease is removed, the lower layer of precipitation is the by-product of degumming, and its main component is phospholipid.

Embodiment 2

[0049] The preparation of hydroxy fatty acid comprises the steps of:

[0050] (1) Dissolve the degummed by-product obtained in Example 1 in acetonitrile, ultrasonically disperse the phospholipids in the degummed by-product, and after dispersion, remove the insoluble precipitate by centrifugation to obtain the organic phase of the substrate.

[0051] (2) Enzyme treatment: Dissolve VpPLD, PLB, UPO, and CHoX in a buffer solution with a pH of 7.0-9.0 to obtain an aqueous phase of a compound enzyme solution, wherein VpPLD and PLB are added at 200 U / g of substrate mass, and UPO is added at The substrate mass was added at 300U / g, CHoX was added at the substrate mass of 350U / g, and the enzyme activity ratio of PLB:VpPLD:UPO:CHoX in the complex enzyme was 5:5:6:7.

[0052] (3) Reaction conditions: The organic phase of the substrate and the aqueous phase of the composite enzyme solution were mixed at a volume ratio of 1:4, and rotated in an oil bath at 30° C. for 2 hours at 500 rpm.

Embodiment 3

[0054] Carry out GC-MS detection to the hydroxy fatty acid mother liquor obtained in embodiment 2.

[0055] Preparation of standard products: after derivatization of existing hydroxy fatty acid standard products, GC-MS detection

[0056] Standard product processing method: 1mL of 25mM hydroxy fatty acid mother solution (dissolved in acetonitrile), add an equal amount of methyl tert-butyl ether for extraction, centrifuge, take the upper organic phase, add anhydrous sodium sulfate to remove water. Then take 400uL of the treated sample in the chromatographic bottle, add 100uL N,O-bis(trimethylsilyl)trifluoroacetamide (containing trimethylchlorosilane) for derivatization reaction (this step is rapid, because the derivatization reagent decomposes with water ), the reaction temperature was 70°C, and the derivatization was performed for 2h.

[0057] The gas phase column and program used in this experiment are as follows:

[0058] Column: Sh-Rxi-5sil-MS column (30*0.25*0.25)

[005...

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Abstract

The invention discloses a method for preparing hydroxy fatty acid. The method comprises the following steps: dissolving degumming byproducts in acetonitrile, carrying out ultrasonic dispersion, removing precipitates by centrifugation to obtain an organic phase, dissolving phospholipase B, phospholipase D, non-specific peroxygenase and choline oxidase in a buffer solution, mixing the organic phase with compound enzyme solution, and reacting for at least 1 hour to obtain the hydroxy fatty acid. The method of the invention provides a new approach for catalytic synthesis of hydroxy fatty acid, realizes cyclic utilization of oil degumming byproducts, and reduces industrial waste.

Description

technical field [0001] The invention belongs to the field of enzyme engineering, and in particular relates to a method for preparing hydroxy fatty acid by combining multiple enzymes. Background technique [0002] Hydroxy fatty acid is an organic compound in which one or several hydrogen atoms in the hydrocarbon chain of a fatty acid are replaced by hydroxyl groups. Its molecules contain both hydroxyl and carboxyl groups. If the hydroxyl and carboxyl groups undergo various chemical reactions, they can be prepared. Metal processing oils, surfactants, cosmetic bases, etc., are widely used. The synthetic methods of hydroxy fatty acid include chemical method and enzymatic method. The chemical method directly replaces the hydrogen atoms in the hydrocarbon chains of fatty acids with hydroxyl groups through substitution reactions. Compared with chemical methods, enzymatic preparation has the advantages of high efficiency, energy saving, clean and environmental protection. The use...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P7/64
CPCC12P7/6409
Inventor 洪淏民杨博王永华马云建蓝东明梁鸿镜
Owner SOUTH CHINA UNIV OF TECH
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