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Method for purifying bromelin by nano-titanium dioxide porous ceramic column chromatography

A technology of nano-titanium dioxide and bromelain, which is applied in the field of separation and purification of bromelain, can solve the problem of low enzyme purity and achieve high-efficiency purification

Inactive Publication Date: 2010-05-19
吴茂玉 +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] In order to solve the technical problem of low enzyme purity in the current production of bromelain, the present invention uses the high adsorption of nano-titanium dioxide to bromelain and the characteristics of high specific surface area of ​​porous ceramics to sinter nano-titanium dioxide into a porous ceramic adsorption material, and through column chromatography Technology realizes efficient purification and separation of bromelain

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0007] According to the ratio of 40% nano-titanium dioxide, 30% clay, 10% quartz, 10% activated carbon, and 10% feldspar, the raw materials are mixed evenly, mixed with water, shaped, dried, and calcined at 1200 ° C for 1 hour to make it porous Ceramic material. Sieve the material so that the particle size is 1-2 mm, rinse with distilled water to remove fine powder, and pack the column by wet method. Crush the pineapple pomace, squeeze the juice, filter, and centrifuge at 3000r / min to obtain the crude enzyme. The pH of the bromelain crude extract was adjusted to 4.6, and the adsorption was carried out at 25°C, and the flow rate of the enzyme solution was 1.5BV / h. A pH 5.8, 0.4 mol / L citrate buffer was used as the elution buffer, and the flow rate of the elution buffer was 1 BV / h. Under the operating conditions of 0.3Mpa and 25°C, the eluate is first ultrafiltered with an ultrafiltration membrane with a cut-off value of 50KD to remove impurities with a large molecular weight;...

Embodiment 2

[0009] According to the ratio of 38% nano-titanium dioxide, 30% clay, 10% quartz, 12% activated carbon, and 10% feldspar, the raw materials are mixed evenly, mixed with water, shaped, dried, and calcined at 1200 ° C for 1 hour to make a porous ceramic material . Sieve the material so that the particle size is 1-2 mm, rinse with distilled water to remove fine powder, and pack the column by wet method. Crush the pineapple, squeeze the juice, filter, and centrifuge at 3000r / min to obtain the crude enzyme. The crude bromelain was dissolved, the pH of the crude bromelain extract was adjusted to 4.7, and the adsorption was carried out at 15° C., and the flow rate of the enzyme solution was 1.5 BV / h. Use pH6.0, 0.4mol / L citrate buffer as the elution buffer, and the flow rate of the elution buffer is 1BV / h. Under the operating conditions of 0.2Mpa and 25°C, the eluate is first ultrafiltered with an ultrafiltration membrane with a cut-off value of 50KD, and then the impurity with a s...

Embodiment 3

[0011] According to the ratio of 42% nano-titanium dioxide, 29% clay, 10% quartz, 10% activated carbon, and 9% feldspar, the raw materials are mixed evenly, mixed with water, shaped, dried, and calcined at 1200 ° C for 1 hour to make a porous ceramic material . Sieve the material so that the particle size is 1-2 mm, rinse with distilled water to remove fine powder, and pack the column by wet method. Crush the pineapple stems, squeeze the juice, filter, and centrifuge to obtain crude enzymes. The crude bromelain was dissolved, the pH of the crude bromelain extract was adjusted to 4.7, and the adsorption was carried out at 20° C., and the flow rate of the enzyme solution was 1.5 BV / h. A pH 5.9, 0.4 mol / L citrate buffer was used as the elution buffer, and the flow rate of the elution buffer was 1 BV / h. Under the operating conditions of 0.25Mpa and 25°C, the eluate is ultrafiltered with an ultrafiltration membrane with a cut-off value of 50KD, and small molecular impurities are ...

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Abstract

The invention discloses a method for purifying bromelin by a nano-titanium dioxide porous ceramic column chromatography, belonging to the technical field of enzyme separation and purification, mainly solving the problems of bromelin purification and enzymatic activity preservation. The technical scheme comprises the following steps of: manufacturing a porous ceramic column chromatography stuffing by a utilizing ceramic sintering technology; purifying the bromelin by adopting a column chromatography technology; and maximally preserving enzymatic activity through ultrafiltration and freeze-drying to obtain a finished product of the bromelin with the amount of activity units reaching 6 million. The invention is a high-efficiency and safe method for preparing the high-activity bromelin, and the product can be used as food or medicine raw material to be widely applied to the fields of food, biomedical industry and the like.

Description

Technical field: [0001] The invention relates to the technical field of bromelain separation and purification. Background technique: [0002] Bromelain is a kind of proteolytic enzyme that exists in pineapple stems and fruits. It is widely used in the fields of food, pharmaceutical industry and cosmetics. At present, the extraction methods of bromelain mainly include kaolin adsorption method, organic solvent precipitation method and ultrafiltration method, but there are problems such as low purification degree and easy inactivation of enzyme solution in production. Studies have shown that nano-titanium dioxide has a small particle size and density, has a very high specific surface area, and has a good adsorption effect on bromelain. At the same time, titanium dioxide is a safe food coloring, which is safe to use in food processing, harmless to human body, and can withstand the treatment of low-concentration NaOH and HCl solutions, and has good chemical inertness. The inven...

Claims

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Application Information

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IPC IPC(8): C12N9/50C04B35/46C04B38/08C04B35/622
Inventor 吴茂玉宋烨赵岩马超和法涛葛邦国
Owner 吴茂玉
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