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

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Problems solved by technology

Currently, there are no reports of attempts to apply hydrophobic surface-activated magne

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

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Experimental program
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Example Embodiment

[0026] Example 1

[0027] Add 3.0 g FeSO 4 ·7H 2 O, 5.8 g FeCl 3 ·6H 2 Dissolve O in 200 mL of oxygen-free water and sonicate for 30 min to make the solution evenly mixed. The temperature is increased to 85 ℃, and N is stirred at 1 300 rpm. 2 Bubbling for 30 minutes, quickly add 7.5 mL of concentrated ammonia water, keep the temperature, stirring and N 2 After the reaction for 30 minutes, the precipitate obtained by magnetic separation was completed and washed with anaerobic water to pH = 7, and washed with 0.015 mol / L ethanol aqueous solution for 3 times;

[0028] 2) Add 80 mL of 50% ethanol aqueous solution, 8 mL of silane cross-linking agent, stir for 5 h at 50 ℃, 200 rpm, and wash 3 times with pH = 7 phosphate buffer;

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

[0030] ...

Example Embodiment

[0033] Example 2

[0034] 1) Add 0.20 mol of ferric salt and 0.10 mol of ferric salt to 200 mL of oxygen-free water, adjust to pH = 1.7 with hydrochloric acid, and ultrasonically treat for 20 minutes; heat to 65 ℃, and stir at 1,500 rpm. 2 Bubble for 20-30 minutes, add 10 mL of 28% ammonia water, 30 minutes later, magnetically separate, wash with oxygen-free water 10 times, and wash with 0.010 mol / L ethanol aqueous solution twice;

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

[0036] 3) Add 20 mL of pH = 7 phosphate buffer and 20 mL of 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 ℃, 50 mg Mucor javanicus The crude lipase enzyme powder was dissolved in 1 mL pH = 10 phospha...

Example Embodiment

[0039] Example 3

[0040] 1) Add 0.30 mol of ferric salt and 0.15 mol of ferric salt to 200 mL of oxygen-free water, adjust to pH = 1.7 with hydrochloric acid, and ultrasonically treat for 20 min; increase the temperature to 90 ℃, and stir at 1 500 rpm. 2 Bubble for 20-30 minutes, add 10 mL of 28% ammonia water, 15 minutes later, magnetically separate, wash with oxygen-free water 10 times, and wash with 0.010 mol / L ethanol aqueous solution twice;

[0041] 2) Add 80 mL of 50% ethanol aqueous solution, 4 mL of silane cross-linking agent, stir for 6 h at 60 ℃, 180 rpm, and wash 5 times with pH = 7 phosphate buffer;

[0042] 3) Add 16 mL of pH = 7 phosphate buffer and 16 mL of 5% glutaraldehyde aqueous solution, stir at 180 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.

[0043] At 4 ℃, 50 mg Mucor javanicus The crude lipase enzyme powder was dissolved in 1 mL pH = 10 p...

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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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