Method for extracting papain by using double aqueous phase system

A two-phase system, papain technology, applied in the direction of hydrolytic enzymes, biochemical equipment and methods, enzymes, etc., can solve the problems of unstable enzyme activity and cumbersome process, and achieve simple experimental conditions, good chemical stability, and easy Magnified effect

Active Publication Date: 2017-12-19
JIANGSU UNIV
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  • Claims
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Problems solved by technology

However, high-purity papain is mainly used in the pharmaceutical industry, and its source mainly depends on imports. The existing industrial production technology of papain has unstable enzyme activity and complicated process. Therefore, it is important to study the extraction of high-purity protein with two-phase system. Value

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  • Method for extracting papain by using double aqueous phase system
  • Method for extracting papain by using double aqueous phase system

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] (1) Add 2g of polyethylene glycol 400 to a 10ml centrifuge tube, and then add 4g of pre-configured NaH with a mass concentration of 60wt%. 2 PO 4 -2H 2 O solution, followed by the addition of 1g of [C with a mass concentration of 40wt% 2 mim]Cl solution, followed by the addition of 1ml of pre-prepared papain solution with a mass concentration of 10mg / ml, and finally added to 10g with distilled water.

[0021] In order to test the accuracy of the data, each sample was equipped with three parallel experiments.

[0022] (2) After the preparation of the two-phase aqueous system is completed, place it on a vortex shaker for 5 minutes to ensure uniform mixing of the entire system, and then put it in a constant temperature water bath at 25°C and let it stand for 24 hours. After the phase separation was completed, the upper and lower phase solutions were taken respectively to measure the concentration of papain.

[0023] (3) in the present invention, papain concentration me...

Embodiment 2

[0025] (1) Add 2g of polyethylene glycol 400 to a 10ml centrifuge tube, and then add 4g of pre-configured NaH with a mass concentration of 60wt%. 2 PO4 -2H 2 O solution, followed by the addition of 1g of [C with a mass concentration of 40wt% 4 mim]Cl solution, followed by the addition of 1ml of pre-prepared papain solution with a mass concentration of 10mg / ml, and finally added to 10g with distilled water. In order to test the accuracy of the data, each sample was equipped with three parallel experiments.

[0026] (2) After the preparation of the two-phase aqueous system is completed, place it on a vortex shaker for 5 minutes to ensure uniform mixing of the entire system, and then put it in a constant temperature water bath at 25°C and let it stand for 24 hours. After the phase separation was completed, the upper and lower phase solutions were taken respectively to measure the concentration of papain.

[0027] (3) in the present invention, papain concentration measures ultr...

Embodiment 3

[0029] (1) Add 2g of polyethylene glycol 400 to a 10ml centrifuge tube, and then add 4g of pre-configured NaH with a mass concentration of 60wt%. 2 PO 4 -2H 2 O solution, followed by the addition of 1g of [C with a mass concentration of 40wt% 6 mim]Cl solution, followed by the addition of 1ml of pre-prepared papain solution with a mass concentration of 10mg / ml, and finally added to 10g with distilled water. In order to test the accuracy of the data, each sample was equipped with three parallel experiments.

[0030] (2) After the preparation of the two-phase aqueous system is completed, place it on a vortex shaker for 5 minutes to ensure uniform mixing of the entire system, and then put it in a constant temperature water bath at 25°C and let it stand for 24 hours. After the phase separation was completed, the upper and lower phase solutions were taken respectively to measure the concentration of papain.

[0031] (3) in the present invention, papain concentration measures ul...

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Abstract

The invention discloses a method for extracting papain by using a double aqueous phase system, belongs to the technical field of bioseparation, and particularly relates to an ionic liquid assisted polyethylene glycol/sodium dihydrogen phosphate system. The method comprises the following specific steps: taking imidazolium ionic liquid as an auxiliary material, and adding the imidazolium ionic liquid into a polyethylene glycol 400/sodium dihydrogen phosphate double aqueous phase system prepared according to a certain mass fraction; then adding a papain solution with a certain concentration; after preparation is finished, putting the prepared solution on a vortex oscillator, oscillating for 5 minutes and uniformly mixing; subsequently, putting the mixed solution into a thermostat water bath with the temperature of 25 DEG C and allowing standing still for 24 hours; after the phase splitting operation is finished, respectively taking an upper phase and a lower phase, diluting the solution to a certain concentration with distilled water, and measuring the concentration of the papain in each phase. The method disclosed by the invention has the characteristics that the extraction rate of the papain by a polymer/salt double aqueous phase system can be remarkably improved; besides, the viscosity of the double aqueous phase system can be reduced; the method is low in energy consumption, simple in operation and the like.

Description

technical field [0001] The invention belongs to the technical field of biochemical separation, and relates to a method for extracting papain by using a two-phase aqueous system, in particular to a method for extracting papain by using an ionic liquid-assisted polymer / inorganic salt two-phase system. Background technique [0002] Protein is a kind of biological macromolecules that are crucial to life activities. Various life functions, life phenomena, and life activities are all related to proteins. It plays a key role in many aspects of living organisms, such as catalysis, movement, structure recognition and regulation, and participates in every process of cell life activities. Therefore, protein is widely used in industries such as medicine, food, additives and cosmetics, and its demand is increasing, its quality requirements are getting higher and higher, varieties are changing faster and faster, and production process reforms are getting newer. It can be seen that the qu...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/50
CPCC12N9/641C12Y304/22002
Inventor 周存山杨宏朋张磊余筱洁江昊南冯亚斌
Owner JIANGSU UNIV
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