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Method for preparing 1,3-diglyceride from surface active magnetic nanoparticle immobilized lipase

A technology of magnetic nanoparticles and immobilized lipase, which is applied in the direction of immobilization on or in inorganic carriers, fermentation, etc., to achieve the effects of improved activity and operational stability, great application value, and convenient recycling and reuse

Inactive Publication Date: 2012-06-27
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Currently, there are no reports of attempts to apply hydrophobic surface-activated magnetic nanoparticle-immobilized lipase to 1,3-diglyceride synthesis in a solvent-free system

Method used

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  • Method for preparing 1,3-diglyceride from surface active magnetic nanoparticle immobilized lipase
  • Method for preparing 1,3-diglyceride from surface active magnetic nanoparticle immobilized lipase
  • Method for preparing 1,3-diglyceride from surface active magnetic nanoparticle immobilized lipase

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] 3.0 g FeSO 4 ·7H 2 O, 5.8 g FeCl 3 ·6H 2 O was dissolved in 200 mL of anaerobic water and sonicated for 30 min to make the solution evenly mixed, heated to 85 °C, and stirred at 1 300 rpm under N 2 After bubbling for 30 min, quickly add 7.5 mL concentrated ammonia water, keep the temperature, stir and N 2After reacting for 30 min, the precipitate obtained was magnetically separated and washed with anaerobic water to pH = 7, and washed three times with 0.015 mol / L ethanol aqueous solution;

[0028] 2) Add 80 mL of ethanol aqueous solution with a volume fraction of 50%, 8 mL of silane crosslinking agent, stir at 50 °C and 200 rpm for 5 h, and wash with pH = 7 phosphate buffer three times;

[0029] 3) Add 20 mL pH = 7 phosphate buffer solution and 16 mL 5% glutaraldehyde aqueous solution, stir at 200 rpm for 2 h, magnetically separate, wash with pH = 7 phosphate buffer solution for 3 times, and dry in vacuum to obtain Surface activated magnetic nanoparticles.

[003...

Embodiment 2

[0034] 1) Add 0.20 mol of ferric salt and 0.10 mol of ferrous salt to 200 mL of anaerobic water, adjust to pH = 1.7 with hydrochloric acid, and sonicate for 20 min; 2 After bubbling for 20-30 min, add 10 mL of ammonia water with a mass fraction of 28%, after 30 min, magnetically separate, wash with anaerobic water 10 times, and wash with 0.010 mol / L ethanol aqueous solution twice;

[0035] 2) Add 100 mL of ethanol aqueous solution with a volume fraction of 50%, 8 mL of silane cross-linking agent, stir at 50-60 °C and 220 rpm for 6 h, and wash with pH = 7 phosphate buffer for 3-5 times;

[0036] 3) Add 20 mL pH = 7 phosphate buffer and 20 mL 5% glutaraldehyde aqueous solution, stir at 220 rpm for 3 h, magnetically separate, wash 5 times with pH = 7 phosphate buffer, and vacuum dry to obtain Surface-activated magnetic nanoparticle microsphere carrier.

[0037] At 4 °C, 50 mg Mucor javanicus Dissolve lipase crude enzyme powder in 1 mL pH = 10 phosphate buffer, vortex for 90 ...

Embodiment 3

[0040] 1) Add 0.30 mol of ferric salt and 0.15 mol of ferrous salt to 200 mL of anaerobic water, adjust to pH = 1.7 with hydrochloric acid, and sonicate for 20 min; 2 After bubbling for 20-30 min, add 10 mL of ammonia water with a mass fraction of 28%, after 15 min, magnetically separate, wash 10 times with anaerobic water, and wash 2 times with 0.010 mol / L ethanol aqueous solution;

[0041] 2) Add 80 mL of ethanol aqueous solution with a volume fraction of 50%, 4 mL of silane crosslinking agent, stir at 60 °C and 180 rpm for 6 h, and wash with pH = 7 phosphate buffer for 5 times;

[0042] 3) Add 16 mL pH = 7 phosphate buffer solution and 16 mL 5% glutaraldehyde aqueous solution, stir at 180 rpm for 3 h, magnetically separate, wash 5 times with pH = 7 phosphate buffer solution, and vacuum dry to obtain Surface-activated magnetic nanoparticle microsphere carrier.

[0043] At 4 °C, 50 mg Mucor javanicus Dissolve lipase crude enzyme powder in 1 mL pH = 10 phosphate buffer, v...

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Abstract

The invention discloses a method for preparing 1,3-diglyceride from a surface active magnetic nanoparticle immobilized lipase, which comprises the following steps of: (1) preparing a superparamagnetic ferriferrous oxide (Fe3O4) nanoparticle by coprecipitation performed on a mixture of divalent ferric salt and trivalent ferric salt; performing modification by a silane coupling agent; and activating the surface of the modified nanoparticle by glutaraldehyde to obtain a surface active magnetic nanoparticle; (2) adding a lipase solution, stirring, washing and drying to obtain the immobilized lipase; and (3) catalyzing glycerol to perform an esterification reaction with a fatty acid by adopting the immobilized lipase to obtain the 1,3-diglyceride. By utilizing the immobilized lipase disclosed by the invention, the activity, the operation stability and the regioselectivity are obviously improved in the synthetic reaction of the 1,3-diglyceride; the recycling operation of the lipase is greatly simplified; and moreover, the 1,3-diglyceride is high in purity and does not contain harmful solvents.

Description

field of invention [0001] The invention relates to a preparation method of a structure-specific compound, in particular to a method for preparing 1, 3-diglyceride by immobilizing lipase with surface-activated magnetic nanoparticles. Background technique [0002] 1, 3-Diglyceride is a structural analogue of natural edible oil (triglyceride). Its taste and appearance are consistent with the former, but its metabolism and absorption in the human body are different: triglyceride is digested by digestive enzymes to form mono Glycerides and free fatty acids, after the two are absorbed into the blood, a large part of them re-synthesize triglycerides to increase blood lipids or cause fat accumulation. However, 1, 3-diglycerides are processed by digestive enzymes to generate glycerol and free fatty acids. Glycerol enters the TCA cycle through pyruvate, and free fatty acids are transported to the liver for β-oxidation. Therefore, eating oils containing 1,3-diglycerides has the effect...

Claims

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Application Information

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IPC IPC(8): C12P7/62C12N11/14
Inventor 孟枭徐刚吴坚平杨立荣
Owner ZHEJIANG UNIV
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