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Method for preparing magnetically regenerable immobilized bio-enzyme

A biological enzyme and magnetic technology, applied in the field of biological enzyme catalysis, can solve the problems of magnetic materials losing single-domain magnetic poles, loss of dispersibility, easy agglomeration and coagulation, etc., to facilitate recycling and regeneration, improve dispersion and stability sexual effect

Inactive Publication Date: 2013-11-20
SHANDONG NORMAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although magnetic nanoparticles have reached nanometer size, they usually have a wide particle size distribution, bare surface, active chemical properties, and are easily oxidized in the air. Due to the magnetic attraction and van der Waals force, they are prone to agglomeration and coagulation. Phenomena that make magnetic materials lose single-domain magnetic poles, dispersibility, or lose properties unique to nanomaterials

Method used

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  • Method for preparing magnetically regenerable immobilized bio-enzyme
  • Method for preparing magnetically regenerable immobilized bio-enzyme
  • Method for preparing magnetically regenerable immobilized bio-enzyme

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Accurately weigh 13.104g FeSO with an electronic balance 4 ·7H 2 O in a small beaker, add 50ml of deionized water and stir to dissolve; 13.886gNa 2 S 2 o 3 ·5H 2 O in a small beaker, add 25ml of deionized water and stir to dissolve; add 4.480g of powdery NaOH to a small beaker, add 25ml of deionized water and stir to dissolve; after the dissolution is complete, mix NaOH aqueous solution and NaOH 2 S 2 o 3 The aqueous solution was transferred to a 250ml three-neck flask. Then FeSO 4 The aqueous solution was transferred to a constant pressure funnel, and the flow rate was controlled to allow the aqueous solution to drop into a three-necked flask. After the dropwise addition, turn on the electric stirrer to stir for 1 h, then transfer all the reactants in the three-neck flask into a hydrothermal kettle, and react at 160° C. for 16 hours. After the reaction, all the products were transferred to a watch glass, and the supernatant was sucked out after sedimentation w...

Embodiment 2

[0028] Surface modified Fe 3 o 4 The preparation of the granules is the same as in Example 1.

[0029] Weigh 1.0g of α-amylase and dissolve it in a 100ml beaker with buffer solution, stir with a glass rod for a period of time until the solid amylase dissolves, filter with filter paper to remove insoluble impurities, and transfer the obtained clear liquid to a 100ml volumetric flask, Set aside.

[0030] Weigh 0.1g of Fe after surface modification 3 o 4 Put the granules in a 250ml Erlenmeyer flask, add 100ml of buffer solution, add (accounting for the total volume) 1% crosslinking agent glutaraldehyde, crosslink and activate for 20min, measure the prepared α-amylase solution 10ml, added to the activated solution, reacted in a constant temperature oscillator for 2 hours, washed with deionized water for several times, and prepared a nano-magnetic material with α-amylase immobilized. After testing, the amylase activity after immobilization reached 87% of the free enzyme, and t...

Embodiment 3

[0032] Add 5.50g of cetyltetrabutylammonium bromide (CTAB), 16.50g of n-butanol and 68.00g of isooctyl alcohol into a 250ml three-necked flask and stir to form a homogeneous mixture, and add 10ml of 0.1M ferric chloride solution into the mixture , stir vigorously until orange and transparent, and pass through N 2 Protection, then slowly add 0.12g sodium borohydride powder, continue to stir and pass N 2 After the system turns black and no longer emits gas, continue to stir for half an hour. Stand still until all the black powder in the system sinks to the bottom of the bottle, suck out the upper liquid, add 50ml of methanol and chloroform mixed washing solution to wash, and use the same method to suck out the washing solution, then use 50ml of absolute ethanol, and use the same method Washing to obtain nanometer zero-valent ferromagnetic particles.

[0033] Weigh 54g of absolute ethanol, add 3g of deionized water, then add 2.38g of KH570, hydrolyze for about 10min, add 0.3g o...

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Abstract

The invention relates to a method for preparing a magnetically regenerable immobilized bio-enzyme. The method comprises the following steps of firstly preparing ferromagnetism nano particles, mixing absolute ethyl alcohol, deionized water and a silane coupling agent, after hydrolysis, adding the ferromagnetism nano particles, stirring and reacting for 1-2 hours under the protection of nitrogen, adsorbing and settling with a magnet, washing and drying the settled solid to obtain the surface-modified ferromagnetism nano particles, then dropping a cross-linking agent for crosslinking and activating, metering a bio-enzyme preparation and oscillating at constant temperature so as to obtain the magnetically regenerable immobilized bio-enzyme. The magnetic material and the high efficient catalytic characteristics of alpha-amylase and cellulose in the bio-enzyme are combined and play a synergistic effect so as to prepare the magnetically regenerable immobilized bio-enzyme, so that the used bio-enzyme is benefit to being recycled efficiently and regenerated, and the preliminary trial on the two enzymes finds that the preparation method can be expanded to be applied to the immobilization of the whole bio-enzyme field.

Description

technical field [0001] The invention relates to a method for preparing a magnetically regenerated immobilized biological enzyme, which uses a hydrothermal method to prepare nanometer ferric oxide or a microemulsion method to prepare nanometer zero-valent iron, modifies it with a silane coupling agent, and then uses a crosslinking agent Activation, the introduction of biological enzymes to prepare novel immobilized biological enzymes, belongs to the technical field of biological enzyme catalysis. Background technique [0002] As a natural biocatalyst, biological enzymes have attracted more and more attention from scientific and technological workers because of their excellent selectivity and unparalleled high efficiency under mild conditions, and have been widely used in biomedicine, chemical industry, food and textile industry is widely used. However, in the process of using biological enzymes as catalysts, the first challenge encountered is the "inactivation" of biological...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N11/14H01F41/02
Inventor 张其坤赵雷振康俊清丁文武张志德
Owner SHANDONG NORMAL UNIV
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