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Method for refining papain

A papain, nylon membrane technology, applied in biochemical equipment and methods, enzymes, peptidases, etc., can solve the problems of small processing capacity, low operation pressure, inability to reach the pharmaceutical industry, etc., and achieve the effect of improving vitality recovery

Inactive Publication Date: 2016-11-23
TAICANG YAFENG FARM PROFESSIONAL COOP
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, the extraction and purification of papain mainly adopts the precipitation method, but this method is cumbersome, and the product is still mixed with other proteases, the purity is not high, and it cannot meet the needs of the pharmaceutical industry
As people put forward further requirements for purity and safety, chromatographic techniques are applied to the purification of papain, such as ion exchange chromatography and affinity chromatography. Low axial rate limitation, large pressure drop, long separation time, small throughput, cannot use high feed rate
Membrane separation is a separation technology based on the size of molecules. It is characterized by large surface area, short diffusion path, low operating pressure, and large processing capacity. However, traditional membrane separation technology can only separate species with a relative molecular mass difference of more than 10 times. Meet the needs of biochemical separation

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] A method for refining papain, comprising the following steps:

[0019] a. Hydrolyze the nylon membrane in hydrochloric acid. The hydrolysis temperature of hydrochloric acid is 20°C, and the hydrolysis time is controlled at 15 hours. Then immerse in 20ml of formaldehyde solution, add 0.5ml of phosphoric acid, and immerse in 10ml of chitosan solution after the reaction is complete. , drying at room temperature after reaction, the drying temperature is controlled at 40°C, and finally washed with 50ml acetic acid and 50ml deionized water;

[0020] b, utilize ultrasonic wave to pulverize papaya, then carry out rough enzyme separation with the nylon membrane in step a;

[0021] c, using polyethylene glycol and potassium phosphate or methyl cellulose and dextran to perform two-phase extraction to purify the refined enzyme.

Embodiment 2

[0023] A method for refining papain, comprising the following steps:

[0024] a. Hydrolyze the nylon membrane in hydrochloric acid. The hydrolysis temperature of hydrochloric acid is 22°C, and the hydrolysis time is controlled at 17 hours. Then immerse in 20ml of formaldehyde solution, add 0.5ml of phosphoric acid, and immerse in 10ml of chitosan solution after the reaction is complete. , drying at room temperature after reaction, the drying temperature is controlled at 45°C, and finally washed with 50ml acetic acid and 50ml deionized water;

[0025] b, utilize ultrasonic wave to pulverize papaya, then carry out rough enzyme separation with the nylon membrane in step a;

[0026] c, using polyethylene glycol and potassium phosphate or methyl cellulose and dextran to perform two-phase extraction to purify the refined enzyme.

Embodiment 3

[0028] A method for refining papain, comprising the following steps:

[0029] a. Hydrolyze the nylon membrane in hydrochloric acid, the hydrolysis temperature of hydrochloric acid is 23°C, the hydrolysis time is controlled at 20 hours, then immerse in 20ml formaldehyde solution, add 0.5ml phosphoric acid, after the reaction is complete, immerse in 10ml chitosan solution , drying at room temperature after reaction, the drying temperature is controlled at 50°C, and finally washed with 50ml acetic acid and 50ml deionized water;

[0030] b, utilize ultrasonic wave to pulverize papaya, then carry out rough enzyme separation with the nylon membrane in step a;

[0031] c, using polyethylene glycol and potassium phosphate or methyl cellulose and dextran to perform two-phase extraction to purify the refined enzyme.

[0032] The beneficial effect of the present invention is: the separation of crude enzymes, eliminating the need for repeated precipitation steps, so that the recovery of ...

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PUM

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Abstract

The invention discloses a method for refining papain. The method is characterized by comprising the following steps: (a) hydrolyzing a nylon membrane in hydrochloric acid and soaking in a formaldehyde solution; adding phosphoric acid; after complete reaction, soaking in a chitosan solution; drying after room-temperature reaction; and washing with acetic acid and deionized water at last; (b) breaking papaya using ultrasonic wave and performing crude enzyme separation by using the nylon membrane in the step (a); and (c) performing aqueous two-phase extraction by using polyethylene glycol and potassium phosphate or methylcellulose and glucan, and purifying the fine enzyme. The invention has the following beneficial effects: through crude enzyme separation, the step of repeated precipitation is omitted, and the activity recovery is greatly improved; and since the raw materials do not contain harmful substances and no harmful substance is introduced in the operation process, the obtained papain product can be directly applied to deep processing of food.

Description

technical field [0001] The invention relates to a refining method, in particular to a refining method of papain. Background technique [0002] Papain, referred to as papain, also known as papaya enzyme, is a pure natural biological enzyme product refined from the milk of immature papaya fruit by modern bioengineering technology. It is a kind of protease containing thiol (~SH) , has the activity of protease and esterase, and its broad specificity has a strong ability to hydrolyze animal and plant proteins, polypeptides, esters, amides, etc., and can catalyze various proteins to be hydrolyzed into a series of short peptides or amino acids of different lengths. It can also synthesize protein hydrolyzate into proteinoids. Because of its good thermal stability and natural hygiene and safety, it is widely used in food, leather, textile, daily chemical and pharmaceutical industries to improve product quality and grade and reduce costs. [0003] At present, the extraction and purif...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/50
CPCC12N9/63C12Y304/22002
Inventor 张茜茜
Owner TAICANG YAFENG FARM PROFESSIONAL COOP