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Carrier material of immobilized papain and preparation method of carrier material

A technology of papain and carrier materials, applied in the direction of immobilization on or in inorganic carriers, immobilization on/in organic carriers, biochemical equipment and methods, etc., which can solve the loss of enzyme activity, intense reaction, and enzyme leakage Low rate and other issues

Inactive Publication Date: 2017-11-17
苏州凯邦生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The covalent binding method is to immobilize the enzyme on a suitable carrier through a covalent bond method. The enzyme and the carrier produce a chemical interaction, the binding force is strong, the enzyme leakage rate is low, and it has good stability and recycling performance. Intense, easy to lose enzyme activity

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] A preparation method for a carrier material of immobilized papain, comprising the following steps:

[0026] (1) Multi-walled carbon nanotubes and nano-titanium dioxide are mixed, put into planetary grinder, grind 20min, powder is taken out afterwards, oven dry, the powder that makes is dispersed in the aqueous solution of polyethylene glycol-2000, more The mass ratio of wall carbon nanotubes, nano-titanium dioxide, and agarose is 1:2:8, ultrasonically dispersed at 50°C for 45 minutes, and then a silane coupling agent is added dropwise to it, and stirred with a high-shear mixer for 2 hours. The stirring speed is 20000rpm, add agarose powder, heat to 80°C until the agarose dissolves, stir evenly, and ultrasonicate for 4 hours to prepare a mixed solution, the concentration of agarose is 8-12%;

[0027] (2) After mixing liquid paraffin and chloroform, add an emulsifier and stir evenly, wherein the emulsifier is a mixture of Span-80 and Span-60, and the volume ratio of liqui...

Embodiment 2

[0032] A preparation method for a carrier material of immobilized papain, comprising the following steps:

[0033] (1) Multi-walled carbon nanotubes and nano-titanium dioxide are mixed, put into planetary grinder, grind 45min, powder is taken out afterwards, oven dry, the powder that makes is dispersed in the aqueous solution of polyethylene glycol-2000, more The mass ratio of wall carbon nanotubes, nano-titanium dioxide, and agarose is 1:2:8, ultrasonically dispersed at 50°C for 60 minutes, and then a silane coupling agent is added dropwise thereto, and stirred for 3 hours using a high-shear mixer. The stirring speed is 30000rpm, add agarose powder, heat to 90°C until the agarose is dissolved, stir evenly, and ultrasonicate for 5 hours to prepare a mixed solution, the concentration of agarose is 12%;

[0034](2) After mixing liquid paraffin and chloroform, add an emulsifier and stir evenly, wherein the emulsifier is a mixture of Span-80 and Span-60, and the volume ratio of li...

Embodiment 3

[0039] A preparation method for a carrier material of immobilized papain, comprising the following steps:

[0040] (1) Multi-walled carbon nanotubes and nano-titanium dioxide are mixed, put into planetary grinder, grind 20min, powder is taken out afterwards, oven dry, the powder that makes is dispersed in the aqueous solution of polyethylene glycol-2000, more The mass ratio of wall carbon nanotubes, nano-titanium dioxide, and agarose is 1:2:8, ultrasonically dispersed at 50°C for 60 minutes, and then a silane coupling agent is added dropwise to it, and stirred with a high-shear mixer for 2 hours. The stirring speed is 30000rpm, add agarose powder, heat to 80°C until the agarose dissolves, stir evenly, and ultrasonicate for 5 hours to prepare a mixed solution, the concentration of the agarose is 8%;

[0041] (2) After mixing liquid paraffin and chloroform, add an emulsifier and stir evenly, wherein the emulsifier is a mixture of Span-80 and Span-60, and the volume ratio of liqu...

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PUM

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Abstract

The invention discloses a carrier material of immobilized papain and a preparation method of the carrier material. The method comprises the steps as follows: liquid paraffin and trichloromethane are mixed, then an emulsifier is added, the mixture is uniformly stirred and kept at the temperature of 60-70 DEGC, a mixed solution is added to the mixture with stirring, the mixture is continuously stirred for 20-30 min after addition and cooled to the room temperature, a supernatant is removed through centrifugation, prepared microspheres are washed three times with petroleum ether and then washed with water, and the carrier material of the immobilized papain is prepared. The invention further discloses the immobilized papain from the carrier material and a preparation method of the immobilized papain. The carrier material can be firmly combined with the papain, and the papain cannot fall off easily during usage. Besides, the papain can still keep higher activity after being immobilized on the carrier.

Description

technical field [0001] The invention relates to the field of enzyme preparations, in particular to a carrier material for immobilized papain and a preparation method thereof. Background technique [0002] Papain is a sulfhydryl protease widely present in the roots, stems, leaves and fruits of the dicotyledonous plant Papain, among which the content of fruit milk is the most abundant. Papain has a good degradation effect on a variety of proteins. It can catalyze the hydrolysis of peptide bonds, ester bonds, and amide bonds. It is a biological enzyme with a wide range of uses and has important applications in many fields. In the food industry, papain is mainly used for tenderizing meat, preventing beer turbidity, and can be used as a feed additive, etc.; in industrial production, it is mainly used for gelatin manufacturing, cocoon degumming, leather depilation, etc.; It is used for deworming, treating indigestion and various inflammations and edema. [0003] Enzyme immobiliz...

Claims

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Application Information

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IPC IPC(8): C12N11/14C12N11/10
CPCC12N9/641C12N11/10C12N11/14C12Y304/22002
Inventor 冯海燕
Owner 苏州凯邦生物技术有限公司
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