Hollow mesoporous carbon nanosphere immobilized enzyme and preparation method thereof

A technology of nano-carbon spheres and immobilized enzymes, which is applied in the direction of immobilization on or in the inorganic carrier, which can solve the problems that the enzyme is difficult to enter the hole, the enzyme is easy to leak, and the stability is not high enough to avoid friction and shearing force Effects of damage and leakage, improvement of immobilization capacity and stability, improvement of stability and activity

Active Publication Date: 2018-03-06
INST OF OIL CROPS RES CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Carriers such as silica gel directly adsorb and immobilize the enzyme, which is not stable, and the enzyme is easy to fall off, requiring complex surface modification; the pore size should match the size of the enzyme, too large will easily cause the loss of the enzyme, and too small will make it difficult for the enzyme to enter inside the pores, resulting in low solid loading
Patent ZL201110350307.0 discloses a method for immobilizing enzymes on attapulgite, but complex silanization modification on the surface of attapulgite is required, and the enzyme immobilization capacity is not high; patent 102443579A discloses a method for applying amphiphilic A method for preparing immobilized enzymes from porous hollow carbon microspheres. The precursor of the hollow carbon material is yeast, and the size of the carrier material formed by carbonization of yeast is micron-scale, and the specific surface area is not large, resulting in a low enzyme immobilization capacity (<30.23mg / g ), the pore size is greater than 50nm, causing the enzyme to leak easily during use, and the stability is not high enough

Method used

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  • Hollow mesoporous carbon nanosphere immobilized enzyme and preparation method thereof

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Embodiment 1

[0035] A hollow mesoporous nano-carbon sphere immobilized enzyme and a preparation method thereof, comprising the following steps:

[0036] 1) Silica / resorcinol-formaldehyde oligomer (SiO 2 Preparation of @RF): Disperse 96mmol (27.68mL) of tetrapropoxysilane into 640mL of ethanol / water reaction solvent with a volume ratio of 7:1, add 24mL, 25%-28% concentrated ammonia water as a catalyst, and stir for 20min , then add 3.2g resorcinol and 4.48mL formaldehyde, stir and react at 25°C for 24h, after the reaction, wash with distilled water and ethanol three times in turn, and dry at 50°C to obtain silica / resorcinol-formaldehyde oligomerization matter (SiO 2 @RF);

[0037] 2) Silicon dioxide / carbon (SiO 2 @C) Preparation of the composite: SiO obtained after drying 2 @RF was placed in a tube furnace, with high-purity nitrogen as a protective gas, raised from room temperature to 600 °C at a rate of 5 °C / min, and calcined at 600 °C for 5 h to obtain a silica / carbon composite ( SiO...

Embodiment 2

[0043] A hollow mesoporous nano-carbon sphere immobilized enzyme and a preparation method thereof, comprising the following steps:

[0044] 1) Silica / resorcinol-formaldehyde oligomer (SiO 2 Preparation of @RF): Disperse 96 mmol (21.00 mL) of tetraethoxysilane into 640 mL of ethanol / water reaction solvent with a volume ratio of 2:1, add 20 mL of 25% to 28% concentrated ammonia as a catalyst, and stir for 20 min , then add 3.2g resorcinol and 4.48mL formaldehyde, stir and react at 25°C for 24h, after the reaction, wash with distilled water and ethanol three times in turn, and dry at 50°C to obtain silica / resorcinol-formaldehyde oligomerization matter (SiO 2 @RF);

[0045] 2) Silicon dioxide / carbon (SiO 2 @C) Preparation: SiO obtained after drying 2 @RF was placed in a tube furnace, with high-purity nitrogen as a protective gas, raised from room temperature to 800 °C at a rate of 5 °C / min, and calcined at 800 °C for 5 h to obtain a silica / carbon composite ( SiO 2 @C);

[0...

Embodiment 3

[0050] A hollow mesoporous nano-carbon sphere immobilized enzyme and a preparation method thereof, comprising the following steps:

[0051] 1) Silica / resorcinol-formaldehyde oligomer (SiO 2 Preparation of @RF): Disperse 24mmol (5.25mL) of tetraethoxysilane and 72mmol (20.76mL) of tetrapropoxysilane into 640mL of ethanol / water reaction solvent with a volume ratio of 3:1, add 32mL, 25% to 28% concentrated ammonia water was used as a catalyst, stirred for 20 minutes, then added 3.2g resorcinol and 4.48mL formaldehyde, stirred and reacted at 25°C for 24h, after the reaction was completed, washed three times with distilled water and ethanol in sequence, and dried at 50°C to obtain Silica / resorcinol-formaldehyde oligomer (SiO 2 @RF);

[0052] 2) Silicon dioxide / carbon (SiO 2 @C) Preparation: SiO obtained after drying 2 @RF was placed in a tube furnace, with high-purity nitrogen as a protective gas, raised from room temperature to 700 °C at a rate of 5 °C / min, and calcined at 700...

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Abstract

The invention discloses hollow mesoporous carbon nanosphere immobilized enzyme and a preparation method thereof. The hollow mesoporous carbon nanosphere immobilized enzyme comprises enzyme and a hollow mesoporous carbon nanosphere acting as a carrier for immobilizing the enzyme, wherein for the hollow mesoporous carbon nanosphere, the particle diameter is 300-400nm, the specific surface area is 1100-1200m<2>/g, the shell thickness is 20-60nm, the cavity size is 250-350nm, and the aperture size of mesopore is 8-14nm. The preparation method comprises the following steps: (1) preparing silicon dioxide/resorcinol-formaldehyde oligomer (SiO@RF); (2) preparing silicon dioxide/carbon (SiO2@C); (3) preparing the hollow mesoporous carbon nanosphere; and (4) immobilizing the enzyme on the hollow mesoporous carbon nanosphere. The hollow mesoporous carbon nanosphere immobilized enzyme has the advantages of greatly improving the loading capacity and stability of enzyme due to the physical structureand hydrophobic surface, being beneficial to exposing the active site of enzyme, and being more beneficial to adsorbent immobilization and activity retention of enzyme compared with common adsorbingmaterials.

Description

technical field [0001] The invention belongs to the technical field of enzyme immobilization, and relates to an enzyme immobilized by hollow mesoporous nanometer carbon spheres and a preparation method thereof. Background technique [0002] Enzymes are green and efficient biocatalysts. However, the free enzyme has poor stability, low activity, and is difficult to recycle and reuse, which greatly limits its application in industry. Enzyme immobilization refers to a technology that limits natural free enzymes to a certain space and cannot move freely, which greatly improves the catalytic activity and stability of enzymes. At present, immobilized enzymes have been widely used in food, medicine, energy, environment and other fields. The carrier of immobilized enzyme is the main factor affecting its performance. Commonly used supports include porous silicon materials, macroporous resins, molecular sieves, etc. Carriers such as silica gel directly adsorb and immobilize the enz...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N11/14
CPCC12N11/14
Inventor 郑明明黄凤洪董喆时杰汤虎邓乾春
Owner INST OF OIL CROPS RES CHINESE ACAD OF AGRI SCI
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