Preparation method of superparamagnetic carrier for crosslinking and immobilizing lipase

A fat immobilization and superparamagnetic technology, applied in the field of food chemistry, can solve problems such as inability to adapt to lipase interface activation characteristics, functional defects, damage, etc., achieve good magnetic separation performance, and overcome the effect of lipase activity-dependent interface effects

Active Publication Date: 2017-03-01
YUNNAN AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
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Problems solved by technology

Since the immobilized enzyme carrier is embedded in the copolymerization reaction of the oleic acid-coated particles in the enoic acid ester, the porosity provided makes the NaNO in the azidation reaction 2 , initiator potassium persulfate, etc. can treat Fe 3 o 4 Causes damage to the oleic acid-coated particles of the core, resulting in functional defects
[0006] Chinese invention patent CN103012673B provides a method for preparing bioenzyme-immobilized core-shell superparamagnetic polymer microspheres, using SiO with strong light water 2 to Fe 3 o 4 Nanoparticles are completely wrapped, and then the surface is introduced into a polymer with epoxy groups. Although this type of immobilized enzyme can better retain enzyme activity and have certain advantages in loading capacity, it can only be used for water-soluble biochemical enzymes. Fixed, unable to adapt to lipase interfacial activation properties

Method used

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  • Preparation method of superparamagnetic carrier for crosslinking and immobilizing lipase

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] (1) Nano-magnetic microsphere Fe 3 o 4 Preparation of:

[0025] FeCl 3 ·6H 2 O was dissolved in anhydrous ethylene glycol to prepare a solution with a concentration of 0.13mmol / mL, and then press FeCl 3 ·6H 2 The mass ratio of O to the treatment agent is 1:6. Add the treatment agent, wherein the treatment agent is sodium acetate and polyethylene glycol. After adding the treatment agent, the concentration of sodium acetate is 95mg / mL, and then transfer to the polytetrafluoroethylene coated with inert material. In a reactor made of vinyl fluoride, ultrasonically react at a constant temperature of 200°C for 5 hours. After natural cooling, wash with ethanol and deionized water alternately for several times, then wash with deionized water, and then dry in vacuum to obtain nano-magnetic particles. Ball Fe 3 o 4 ;

[0026] (2) Nano-magnetic microsphere Fe3 o 4 The package:

[0027] Nano magnetic microsphere Fe obtained in step (1) 3 o 4 Mix with the NaOH solution w...

Embodiment 2

[0032] (1) Nano-magnetic microsphere Fe 3 o 4 Preparation of:

[0033] FeCl 3 ·6H 2 O was dissolved in anhydrous ethylene glycol to prepare a solution with a concentration of 0.1mmol / mL, and then press FeCl 3 ·6H 2 The mass ratio of O to the treatment agent is 1:5. Add the treatment agent, wherein the treatment agent is sodium acetate and polyethylene glycol. After adding the treatment agent, the concentration of sodium acetate is 120mg / mL, and then transfer to the polytetrafluoroethylene coated with inert material. In a reactor made of vinyl fluoride, ultrasonically react at a constant temperature of 200°C for 6 hours. After natural cooling, first wash with ethanol and deionized water alternately for several times, then wash with deionized water, and then vacuum dry to obtain nano-magnetic particles. Ball Fe 3 o 4 ;

[0034] (2) Nano-magnetic microsphere Fe 3 o 4 The package:

[0035] Nano magnetic microsphere Fe obtained in step (1) 3 o 4 Mix with the NaOH solut...

Embodiment 3

[0040] (1) Nano-magnetic microsphere Fe 3 o 4 Preparation of:

[0041] FeCl 3 ·6H 2 O was dissolved in anhydrous ethylene glycol to prepare a solution with a concentration of 0.15mmol / mL, and then press FeCl 3 ·6H 2 The mass ratio of O to the treatment agent is 1:4. Add the treatment agent, wherein the treatment agent is sodium acetate and polyethylene glycol. After adding the treatment agent, the concentration of sodium acetate is 80 mg / mL, and then transfer to the polytetrafluoroethylene coated with inert material. In the vinyl fluoride reactor, ultrasonically react at a constant temperature of 200°C for 4 hours. After natural cooling, wash with ethanol and deionized water alternately for several times, then wash with deionized water, and then dry in vacuum to obtain nano-magnetic particles. Ball Fe 3 o 4 ;

[0042] (2) Nano-magnetic microsphere Fe 3 o 4 The package:

[0043] Nano magnetic microsphere Fe obtained in step (1) 3 o 4 Mix with the NaOH solution whos...

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Abstract

The invention discloses a method for preparing a super paramagnetic carrier of a cross-linking immobilized lipase. The preparation of the super paramagnetic carrier is realized by preparing a nano-magnetic microsphere Fe3O4, coating the nano-magnetic microsphere Fe3O4, functionally modifying the coated nano-magnetic microsphere Fe3O4. A thermo-sensitive material, namely N-isopropylacrylamide, is introduced into the super paramagnetic carrier prepared according to the method, and the structural performance of the carrier can be changed through temperature control, so that the lipase which is immobilized through the super paramagnetic carrier can get out of the interface performance control to realize direct reactions among alcohols, acids and lipids, and separation of the reaction products is facilitated; moreover, lipase activity control through alpha coverage is facilitated. The super paramagnetic carrier is a super paramagnetic polymer microsphere based on the mode of lipase interface activity action; the lipase can be immobilized through the super paramagnetic polymer microsphere; the immobilized lipase is high in magnetic separation performance; 90% or higher of the lipase activity is kept; the detect that the lipase activity in an enzymatic reaction depends on the interface effect is overcome.

Description

technical field [0001] The invention relates to the technical field of food chemistry, in particular to a preparation method of a superparamagnetic carrier for cross-linking and immobilizing lipase. Background technique [0002] Immobilized lipase is the combination of lipase with some object through physical or chemical methods to prepare a complex that is insoluble in water but still has catalytic activity. This process is to convert the free lipase of the monomer into the A complex lipase catalyst bound to an object. [0003] Lipase (EC3.1.1.3) exists widely in nature. Lipase in the industrial field is mainly used in chemical industry, medicine, food, etc., and is derived from microorganisms. In the enzyme-catalyzed reaction, lipase has the characteristics of "interfacial activity" at the oil-water interface. The common catalytic active center, namely the triplet composed of Ser, Asp (Glu) and His, is embedded in one or several tryptophan-containing Under the protection...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C08F120/54C08F2/44C08K9/04C08K9/06C08K3/22C12N11/08
Inventor 刘晓慧张维吕才有杨金玲
Owner YUNNAN AGRICULTURAL UNIVERSITY
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