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Preparation method of immobilized pectinase enzyme taking monodisperse porous polystyrene micro-sphere as carrier

A technology of porous polystyrene, immobilized pectinase, applied in biochemical equipment and methods, immobilized on/in organic carriers, glycosylase, etc., can solve the problem of large loss of activity and poor biocompatibility and other problems, to achieve the effect of small loss of activity, good stability and improved biocompatibility

Active Publication Date: 2017-12-01
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 porous microspheres prepared by this method are monodisperse and have a porous structure. The surface is mainly composed of benzene ring groups, which are hydrophobic and can effectively adsorb enzymes, but their biocompatibility is poor. , resulting in a greater loss of enzyme activity

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] A preparation method of immobilized pectinase with monodisperse porous polystyrene microspheres as a carrier, comprising the following steps:

[0029] (1) Mix and stir polyvinyl alcohol, sodium phosphate and deionized water until the solid is dissolved, and then transfer to a three-necked flask; wherein, the mass ratio of polyvinyl alcohol, sodium phosphate and deionized water is 1:0.35:35;

[0030] (2) Mix and stir azobisisobutyronitrile and styrene, drop them into a three-necked flask, stir for 30 minutes under nitrogen protection, slowly raise the temperature to 65-75°C, reflux for 8 hours, cool to room temperature after the reaction, and centrifuge , the solid was washed with absolute ethanol and dried to obtain polystyrene bulbs; wherein, the mass ratio of polyvinyl alcohol, azobisisobutyronitrile and styrene was 1:0.07:30, and azobisisobutyronitrile and benzene After ethylene is mixed and stirred, the drop rate when it is added dropwise to the three-necked flask i...

Embodiment 2

[0036] A preparation method of immobilized pectinase with monodisperse porous polystyrene microspheres as a carrier, comprising the following steps:

[0037] (1) Mix and stir polyvinyl alcohol, sodium phosphate and deionized water until the solid is dissolved, and then transfer to a three-necked flask; wherein, the mass ratio of polyvinyl alcohol, sodium phosphate and deionized water is 1:0.55:35;

[0038](2) Mix and stir azobisisobutyronitrile and styrene, drop them into a three-necked flask, stir for 30 minutes under nitrogen protection, slowly raise the temperature to 65-75°C, reflux for 11 hours, cool to room temperature after the reaction, and centrifuge , the solid was washed with absolute ethanol and dried to obtain polystyrene bulbs; wherein, the mass ratio of polyvinyl alcohol, azobisisobutyronitrile and styrene was 1:0.12:50, and azobisisobutyronitrile and benzene After ethylene is mixed and stirred, the drop rate when it is added dropwise to the three-necked flask i...

Embodiment 3

[0044] A preparation method of immobilized pectinase with monodisperse porous polystyrene microspheres as a carrier, comprising the following steps:

[0045] (1) Mix and stir polyvinyl alcohol, sodium phosphate and deionized water until the solid is dissolved, and then transfer to a three-necked flask; wherein, the mass ratio of polyvinyl alcohol, sodium phosphate and deionized water is 1:0.4:35;

[0046] (2) Mix and stir azobisisobutyronitrile and styrene, drop them into a three-necked flask, stir for 30 minutes under nitrogen protection, slowly raise the temperature to 65-75°C, reflux for 9 hours, cool to room temperature after the reaction, and centrifuge , the solid was washed with absolute ethanol and dried to obtain polystyrene bulbs; wherein, the mass ratio of polyvinyl alcohol, azobisisobutyronitrile and styrene was 1:0.08:35, and azobisisobutyronitrile and benzene After ethylene is mixed and stirred, the drop rate when it is added dropwise to the three-necked flask is...

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Abstract

The invention discloses a preparation method of immobilized pectinase enzyme taking a monodisperse porous polystyrene micro-sphere as a carrier, and relates to the field of immobilized enzyme preparation. The method includes the steps: preparing the monodisperse porous polystyrene micro-sphere in a seed swelling polymerization manner, and adding an appropriate amount of phthaloyl chitosan when a polystyrene bulb swells to obtain a chitosan modified monodisperse porous polystyrene micro-sphere; mixing and oscillating the chitosan modified monodisperse porous polystyrene micro-sphere, pectinase enzyme water solution and buffer solution to prepare the immobilized pectinase enzyme. The immobilized pectinase enzyme prepared by the method is low in enzyme activity loss, wide in application condition range, good in stability, high in reuse rate and not easy to fall off.

Description

Technical field: [0001] The invention relates to the field of preparation of biological enzyme preparations, in particular to a preparation method of immobilized pectinase using monodisperse porous polystyrene microspheres as a carrier. Background technique: [0002] Pectinase is a kind of enzyme obtained by fermenting and refining Aspergillus niger and used to catalyze the hydrolysis of plant cell interstitium, i.e. pectin, into galacturonic acid and pectinic acid. It is mainly used in the squeezing of fruit and vegetable juice drinks and fruit wine And clarification, it has a good effect on decomposing pectin substances. In my country's fruit juice and wine manufacturing industry, the addition of pectinase can improve the juice yield and stability of fruit and vegetable juices, and prevent the occurrence of turbidity in fruit and vegetable juices. However, in actual industrial applications, the use of free enzymes has disadvantages such as large enzyme consumption, high p...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N11/10C12N11/08C08F291/08C08F212/08C08J9/10
CPCC08F251/00C08F257/02C08J9/102C08J2351/00C08J2351/02C12N9/18C12N9/2408C12N9/88C12N11/08C12N11/10C12Y301/01011C12Y302/01015C12Y402/02002C08F212/08
Inventor 冯海燕
Owner 苏州凯邦生物技术有限公司
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