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Method for preparing silicon base mesoporous molecular sieve SBA-15 immobilized enzyme biocatalyst

A biocatalyst and silicon-based mesoporous technology, which is applied in the direction of immobilization on/in the organic carrier, can solve the problems of easy leakage of immobilized enzymes, unsuitable catalysis of macromolecular substrate reactions, diffusion limitations, etc., and achieve the preparation process Simple and reliable, good enzyme dispersion, good storage stability

Inactive Publication Date: 2009-06-10
SHANGHAI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Traditional enzyme immobilization carriers mainly include the following categories: (1) Inorganic compounds such as kaolin, silica gel, bentonite, alumina, calcium phosphate gel, and microporous glass. The disadvantage of this type of carrier is that the binding force between the enzyme and the carrier is weak , have strict requirements on pH, salt concentration, temperature and other conditions; (2) glutaraldehyde and bis-linked benzidine-2,2'-disulfonic acid, etc., using such carriers to prepare immobilized enzymes, the preparation process is more difficult, and Enzymes cannot be regenerated; (3) polyacrylamides, using polyacrylamide as an embedding agent, the immobilized enzymes prepared by this method are prone to leakage, often have diffusion limitations, catalytic reactions are affected by mass transfer resistance, and are not suitable for catalyzing macromolecules Substrate reactions, etc.

Method used

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  • Method for preparing silicon base mesoporous molecular sieve SBA-15 immobilized enzyme biocatalyst
  • Method for preparing silicon base mesoporous molecular sieve SBA-15 immobilized enzyme biocatalyst
  • Method for preparing silicon base mesoporous molecular sieve SBA-15 immobilized enzyme biocatalyst

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] See attached figure 1 process flow chart. Firstly, the mesoporous molecular sieve SBA-15 carrier was prepared. Polyoxyethylene ether-polyoxypropylene ether-polyoxyethylene ether triblock copolymer (P123) was used as a template, and tetraethyl orthosilicate (TEOS) was used as a silicon source. Weigh 2.0g of P123 into a beaker, add 15g of water and 60g of 2M hydrochloric acid solution, stir at 35°C until P123 is completely dissolved, then add 4.25g of TEOS, and keep stirring at 35°C for 20h. Subsequently, the reactant was transferred to a polytetrafluoroethylene-lined reactor and reacted at 80°C for 24 hours. Finally, the solid product was collected, washed with water, dried at room temperature, and the temperature was slowly raised to 500° C. within 8 hours, and calcined at a constant temperature for 6 hours. The calcined powder is SBA-15 with channels. The prepared SBA-15 has a pore size of about 6.7nm and a specific surface area of ​​686m 2 / g, the specific pore v...

Embodiment 2

[0028] Step a in this example is the same as step a in Example 1; then add 0.2 g of the prepared silicon-based mesoporous molecular sieve SBA-15 carrier to 15 ml of disodium hydrogen phosphate-dihydrogen phosphate with an adsorption reaction pH of 8 Sodium buffer solution, placed on a magnetic stirrer and stirred for 1h to disperse evenly in the solution. After stirring, 1 mg, 2 mg, 4 mg, 6 mg, 8 mg, and 10 mg of horseradish peroxidase were added into the solution and stirred, and the adsorption reaction time was 6 hours. Then filter, wash the molecular sieve material alternately and repeatedly with secondary water and phosphate buffer solution, and finally dry the obtained enzyme-loaded molecular sieve naturally to obtain the immobilized enzyme biocatalyst. attached by Figure 5 It can be seen that, under other conditions being the same, the relative activity of the immobilized enzyme increases as the amount of the enzyme increases. When the amount of the enzyme per gram of ...

Embodiment 3

[0030] Step a in this example is the same as step a in Example 1; then add 0.2 g of the prepared silicon-based mesoporous molecular sieve SBA-15 carrier to 15 ml of disodium hydrogen phosphate-dihydrogen phosphate with an adsorption reaction pH of 8 Sodium buffer solution, placed on a magnetic stirrer and stirred for 1h to disperse evenly in the solution. After the stirring is completed, 4 mg of horseradish peroxidase is added into the solution and stirred, and the adsorption reaction time is 1 h, 2 h, 3 h, 4 h, 5 h, 6 h, 7 h, 8 h. Then filter, wash the molecular sieve material alternately and repeatedly with secondary water and phosphate buffer solution, and finally dry the obtained enzyme-loaded molecular sieve naturally to obtain the immobilized enzyme biocatalyst. attached by Figure 6 It can be seen that the relative enzyme activity of the immobilized enzyme increases with the increase of the adsorption time. When the adsorption time is 4 hours, the relative enzyme activ...

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Abstract

The invention relates to a method for preparing an immobilized enzyme biocatalyst by using silicon-based mesoporous molecular sieve SBA-15, which belongs to the technical field of preparing the immobilized enzyme biocatalyst. The method comprises the steps of taking the silicon-based mesoporous molecular sieve SBA-15 as a carrier, taking horseradish peroxidase as an immobilization object, taking an adsorption method as an immobilization method and utilizing the attribute of the mesoporous molecular sieve and the characteristic that the aperture of the SBA-15 is matched with the molecular size of the horseradish peroxidase to realize the immobilization of enzymes simultaneously on a pore canal and the outer surface of the silicon-based mesoporous molecular sieve SBA-15. The immobilized enzyme biocatalyst prepared by the method has the advantages of good enzyme dispersibility, high enzyme carrying capacity and good enzyme activity. The method can also apply to the immobilization of protease, lipase and a plurality of enzymes.

Description

technical field [0001] The invention relates to a method for preparing an immobilized enzyme biocatalyst using silicon-based mesoporous molecular sieve SBA-15, and focuses on the mechanism analysis of the immobilization process, belonging to the technical field of immobilized enzyme biocatalyst preparation. Background technique [0002] In 1992, Kresge and Beck of Mobil Corporation used surfactants as templates for the first time to synthesize a novel ordered mesoporous silica material MCM-41 (Mobile Composition Material 41), which was a leap in the history of the development of molecular sieves and porous materials. Due to the broad application prospects in the fields of heavy oil catalysis and macromolecule separation, mesoporous molecular sieves have become one of the research hotspots, and a series of mesoporous materials have been developed soon, such as SBA series, MSU series, CMK series, HMS, KIT , and metal and metal oxide series, etc. Mesoporous molecular sieves ar...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N11/08
Inventor 徐佳佳胡龙兴党松涛
Owner SHANGHAI UNIV
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