Preparation method and application of nano material monolithic column immobilized enzyme biological micro-reactor

A technology of nanomaterials and microreactors, applied in enzyme production/bioreactors, methods of supporting/immobilizing microorganisms, biochemical equipment and methods, etc., can solve slow mass transfer rate, low catalytic efficiency, poor service life, etc. question

Inactive Publication Date: 2014-12-10
BEIJING UNIV OF CHEM TECH
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Problems solved by technology

[0005] One of the purposes of the present invention is to study a nano-material monolithic column-immobilized enzyme microbial microreactor, by organically combining the nano-material and the monolithic column, the slow mass transfer rate and enzyme-immobi

Method used

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  • Preparation method and application of nano material monolithic column immobilized enzyme biological micro-reactor

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Embodiment 1

[0026] Embodiment 1: the synthesis of nano gold

[0027] (1) Synthesis of gold nanoparticles with different particle sizes

[0028] Add 50mg of chloroauric acid and 500mL of ultrapure water into a 2000mL Erlenmeyer flask, heat to 98°C under magnetic stirring, then add a certain amount of sodium citrate, keep the temperature at 98°C for reaction, and the color of the reaction solution will last for a few seconds Turn blue, then black, then red, continue heating for 10 min, stop heating, continue stirring for 0.5 h, prepare nano-gold colloid solution, cool at room temperature, high-speed centrifugal purification, save for later use. The particle size of the synthesized gold nanoparticles was changed by changing the amount of sodium citrate, as shown in Table 1. The particle size and morphology of the synthesized gold nanocolloids were studied by transmission electron microscopy (TEM) ( figure 1 ).

Embodiment 2

[0029] Example 2: Fe 3 o 4 SiO 2 -NH 2 Synthesis of Magnetic Nanoparticles

[0030] (1) Solvothermal Synthesis of Magnetic Fe 3 o 4 nanoparticles

[0031] FeCl 3 .6H 2 O (1.35g, 5mmol) and NaAc (3.6g) were dissolved in ethylene glycol (50mL), sonicated for 30min to fully dissolve and disperse the solid, and the dispersion was transferred to a stainless steel autoclave lined with polytetrafluoroethylene and sealed. React at 200°C for 8 hours, cool naturally after the reaction, wash 4 times alternately with absolute ethanol and water with the help of a magnet, and dry in vacuum at 60°C for 6 hours to obtain magnetic Fe 3 o 4 Nanoparticles, whose transmission electron micrographs are shown in figure 2 shown.

[0032] (2) Fe 3 o 4 SiO 2 Synthesis of Core-Shell Structured Nanoparticles

[0033] The magnetic Fe prepared in (1) 3 o 4 350 mg of nanoparticles were dispersed in 160 mL of ethanol, and ultrasonically treated for 0.5 h. Add 1mL ammonia water (25wt%), 40m...

Embodiment 3

[0036] Example 3: Synthesis of metal organic framework material MIL-101

[0037] Utilize the hydrothermal method to synthesize MIL-101, the specific steps are as follows: Weigh 2.0g of Cr(NO 3 ) 3 .6H 2 O was dissolved in 25 mL of deionized water, and after fully dissolved, 0.83 g of terephthalic acid was added, followed by 410 μL of hydrofluoric acid. Then the mixed solution was transferred into a 100 mL stainless steel autoclave lined with polytetrafluoroethylene, and placed in an oven at 220° C. for 8 h. After cooling, the product was filtered, washed, and vacuum-dried at 60°C to obtain a mixture of green powder and white needle-like crystals. The product uses N,N'-dimethylformamide, chloroform and absolute ethanol in sequence to remove white needle-like crystals and unreacted inorganic and organic impurities blocked in the pores to obtain MIL with a particle size of about 500nm -101. Using the BET method to measure the specific surface area of ​​MIL-101, it is 3000m ...

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Abstract

The invention discloses a preparation method and an application of a nano material monolithic column immobilized enzyme biological micro-reactor. The preparation method comprises the following steps of firstly, preparing a porous organic polymer monolithic column by using a mixed solution of a functional monomer, a crosslinking agent, a pore forming agent and an initiating agent through in-situ thermal-initiated or light-initiated polymerization in the column, and then bonding a nano material after functional modification to obtain a nano material monolithic column; and secondly, realizing immobilization of the enzyme on the monolithic column by using the nano material as an intermediate ligand to obtain the nano material monolithic column immobilized enzyme biological micro-reactor. The biological micro-reactor is successfully applied to proteomic analysis, medicament chiral resolution and catalyzed ester exchange reactions. The biological micro-reactor disclosed by the invention has the following advantages that the preparation method is simple, the immobilization amount of the enzyme is large, the catalytic activity is high, the enzymolysis speed is high, the efficiency is high, the service life is long, and the biological micro-reactor can be reused.

Description

technical field [0001] The present invention relates to the preparation and application of a monolithic column-immobilized enzyme biomicroreactor, in particular to a method for preparing a porous organic polymer monolithic column-immobilized enzyme biomicroreactor based on nanomaterials as an intermediate ligand, and to study its Applications in proteomic analysis, chiral resolution of drugs and catalytic transesterification reactions. Background technique [0002] Enzyme immobilization technology is an interdisciplinary technology. Immobilized enzymes have higher enzyme / substrate ratio and enzymatic hydrolysis efficiency, faster enzymatic hydrolysis rate, can be used continuously and can reduce the enzyme self The advantages of degradation and cross-infection; and the enzymatic hydrolysis reaction is controllable, the enzyme and the product are easy to separate, the product is easy to purify, the space required for the reaction is small, and it can be used in conjunction w...

Claims

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Application Information

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IPC IPC(8): C12M1/40
CPCC12M21/18C12M23/16C12M23/20C12M25/00
Inventor 吕永琴谭天伟王红霞朱正礼林祥华
Owner BEIJING UNIV OF CHEM TECH
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