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Nanometer carbonic anhydrase grain for biological catalysis of polymer and its prepn process

A nano-polymer, carbonic anhydrase technology, applied in the direction of enzyme stabilization, can solve the problems of insignificant improvement of chemical modification stability effect, high mass transfer resistance of immobilized carbonic anhydrase, poor thermal stability, etc., and achieves inhibition of direct hydrophobicity. Interactions and electrostatic interactions, enhancing rigidity, preventing deactivation problems

Active Publication Date: 2007-01-03
TSINGHUA UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] One of the purposes of the present invention is to provide a nano-polymer biocatalytic particle whose core is carbonic anhydrase, to solve the problems of poor thermal stability and easy aggregation of existing carbonic anhydrase, and to further improve the application level of carbonic anhydrase important meaning
[0005] Another object of the present invention is to provide a kind of preparation method of the nano-polymer biocatalytic particle of carbonic anhydrase, to solve existing carbonic anhydrase chemical modification stability effect to improve not obviously and immobilized carbonic anhydrase to introduce higher problem The problem of mass transfer resistance

Method used

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  • Nanometer carbonic anhydrase grain for biological catalysis of polymer and its prepn process
  • Nanometer carbonic anhydrase grain for biological catalysis of polymer and its prepn process
  • Nanometer carbonic anhydrase grain for biological catalysis of polymer and its prepn process

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Embodiment 1

[0034]Example 1: The raw material is 10 parts by weight of bovine carbonic anhydrase, the enzyme modifier is 2 parts by weight of succinimide acrylate, and the alkenyl monomer is 18 parts of acrylamide and 4.5 parts of methylenebis Acrylamide, a total of 22.5 parts, the initiator is 2 parts of ammonium persulfate and 3 parts of N, N, N', N'-tetramethylethylenediamine, a total of 5 parts. The above-mentioned bovine carbonic anhydrase and enzyme modifier were added into 20 mM boric acid buffer solution with a pH of 8.5, reacted at 4° C. for 2 hours, and dialyzed with water to remove the modifier and borate. Add the obtained mixture to 1% aqueous solution of dimethyl sulfoxide, add 50% of the amount of alkenyl monomer, keep the temperature at 10°C, stir magnetically, add the initiator, react for 0.5 hours, add the remaining alkenyl monomer Keep the temperature at 10°C, continue to react for 2 hours, then use deionized water to dialyze for 12 hours, freeze-dry to obtain bovine car...

Embodiment 2

[0037] Embodiment 2: The raw material is 10 parts by weight of bovine carbonic anhydrase, the enzyme modifier is 5 parts by weight of acryloyl chloride, and the alkenyl monomer is 60 parts of acrylamide and 15 parts of methylenebisacrylamide. 75 parts, the initiator is 5 parts of ammonium persulfate and 10 parts of N, N, N', N'-tetramethylethylenediamine, a total of 15 parts. The above-mentioned bovine carbonic anhydrase and enzyme modifier were added to 200 mM sodium phosphate buffer solution with a pH of 7.5, reacted at 4° C. for 2 hours, and dialyzed with water to remove the modifier and phosphate. The resulting mixture was added to 1% acetonitrile aqueous solution, 20% of the amount of vinyl monomer was added, the temperature was maintained at 10 °C, magnetic stirring was added, the initiator was added, and the reaction was carried out for 0.5 hours, the remaining vinyl monomer was added, and the temperature was maintained The temperature is 10°C, and the reaction is conti...

Embodiment 3

[0038] Embodiment 3: Change the enzyme modifier in embodiment 2 to 10 parts of itaconic anhydride, the vinyl monomer is 80 parts of acrylamide and 20 parts of trihydroxymethylpropane trimethacrylate, free radical reaction solution 4% dioxane, the reaction temperature is 25°C, the initiator is 10 parts of potassium persulfate and 5 parts of sodium bisulfite, all the other formulas and steps are the same as in Example 2, and the stability of the product is obtained now , Activity yield and polymerization yield are equivalent to Example 2, and the product particle diameter is 45~55nm.

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Abstract

The nanometer carbonic anhydrse grain for biological catalysis of polymer and its preparation process belongs to the field of chemical modification of enzyme. The nanometer carbonic anhydrse grain with bioactivity is a core-shell structure with core of carbonic anhydrse, shell of crosslinked polymer material and chemical bond connection between the core and the shell, and possesses high catalytic activity, high heat stability and no aggregation. This kind of nanometer carbonic anhydrse grain has wide application foreground in CO2 absorption and separation and artificial lung system, etc. Its preparation process includes introducing C-C double bond radical with enzyme modifier to the surface of carbonic anhydrse and subsequent free radical polymerization with the olefin monomer with C-C double bond as material.

Description

technical field [0001] The invention relates to a nano-polymer biocatalytic particle whose core is carbonic anhydrase and a preparation method thereof, belonging to the field of chemical modification of enzymes. Background technique [0002] Carbonic anhydrase (carbonic anhydrase) is a zinc-containing metalloprotein that widely exists in organisms. It can efficiently catalyze the hydration and dehydration of carbon dioxide, and is one of the most important enzymes in the carbon dioxide cycle in organisms. In plants or algae, carbonic anhydrase mainly converts carbon dioxide into bicarbonate or carbonate form for photosynthesis through hydration, and in animals mainly converts carbon dioxide in the form of bicarbonate or carbonate in the lungs It is released into the air by dehydration. As a high-efficiency biocatalyst, carbonic anhydrase can increase the absorption or desorption rate of carbon dioxide by more than 2000 times, and can also achieve efficient absorption of ext...

Claims

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Application Information

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IPC IPC(8): C12N9/96
Inventor 闫明刘铮卢滇楠戈钧张敏莲欧阳平凯
Owner TSINGHUA UNIV
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