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Efficient preparation method of papain preparation

A papain and papain technology, applied in biochemical equipment and methods, enzymes, peptidases, etc., can solve problems such as waste, high water and electricity consumption, and difficulty in adapting to industrialized preparation.

Pending Publication Date: 2021-05-18
HAINAN UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the operation of separating and purifying papaya lipase in the prior art is relatively complicated. The more commonly used extraction method of CPL is mainly to remove the water-soluble proteases in papaya milk by repeated washing with buffer, and to reclaim CPL by centrifugation. The yield of CPL is papaya About 2.50% of milk weight, lipase activity is about 350U / g
Although this method can completely remove the water-soluble enzymes in papaya milk, it needs a lot of water and electricity, and the processing capacity is small. Generally, it is only suitable for laboratory and sample trial production, and it is difficult to meet the requirements of industrial production.
At the same time, this method regards the active ingredients of water-soluble proteases as waste, resulting in a large waste of resources

Method used

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  • Efficient preparation method of papain preparation

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Embodiment 1 of the present invention provides a kind of high-efficiency preparation method of papaya enzyme preparation, and it comprises the following specific operation steps:

[0032] S1, the preparation of papaya enzyme preparation crude product

[0033] Select the fresh papaya collected on the same day to make papaya pulp, and add solid sodium sulfite to the papaya pulp. Adjust the pH to 5.5, and freeze-dry the obtained solution to obtain the crude product of the papaya enzyme preparation;

[0034] S2, preparation of papain crude enzyme product solution and papain lipase crude enzyme product

[0035] Dissolve the crude papaya enzyme preparation prepared in step S1 in L-cysteine ​​solution at 4°C, wherein every 90g of the papain lipase preparation is dissolved in 500mL of L-cysteine ​​solution; using an automatic stirrer After stirring and mixing for 30 minutes, the resulting mixture was centrifuged at a speed of 8000 rpm and the centrifugation time was 20 minute...

Embodiment 2

[0042] Embodiment 2 of the present invention provides a kind of high-efficiency preparation method of papaya enzyme preparation, and it comprises the following specific operation steps:

[0043] S1, the preparation of papaya enzyme preparation crude product

[0044]Select the fresh papaya collected on the same day to make papaya pulp, and add solid sodium sulfite to the papaya pulp. Adjust the pH to 6.0, and freeze-dry the obtained solution to obtain the crude product of the papaya enzyme preparation;

[0045] S2, preparation of papain crude enzyme product solution and papain lipase crude enzyme product

[0046] Dissolve the crude papaya enzyme preparation prepared in step S1 in L-cysteine ​​solution at 4°C, wherein every 90g of the papain lipase preparation is dissolved in 500mL of L-cysteine ​​solution; using an automatic stirrer After stirring and mixing for 50 minutes, the resulting mixed solution was centrifuged at a speed of 12000 rpm and the centrifugation time was 10...

Embodiment 3

[0053] Embodiment 3 of the present invention provides a kind of high-efficiency preparation method of papaya enzyme preparation, and it comprises the following specific operation steps:

[0054] S1, the preparation of papaya enzyme preparation crude product

[0055] Select the fresh papaya collected on the same day to make papaya pulp, and add solid sodium sulfite to the papaya pulp. Adjust the pH to 6.0, and freeze-dry the obtained solution to obtain the crude product of the papaya enzyme preparation;

[0056] S2, preparation of papain crude enzyme product solution and papain lipase crude enzyme product

[0057] Dissolve the crude papaya enzyme preparation prepared in step S1 in L-cysteine ​​solution at 4°C, wherein every 90g of the papain lipase preparation is dissolved in 500mL of L-cysteine ​​solution; using an automatic stirrer After stirring and mixing for 40 minutes, the resulting mixture was centrifuged at a speed of 10,000 rpm and the centrifugation time was 10 minu...

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Abstract

The invention relates to an efficient preparation method of a papain preparation. The method comprises the following specific operation steps: S1, preparing a papain preparation crude product; S2, preparing a papain crude enzyme product solution and a papaya lipase crude enzyme product; S3, preparing a papaya lipase preparation; and S4, isolating and purifying papain. According to the efficient preparation method of the papain preparation disclosed by the embodiment of the invention, the papain and the papaya lipase can be obtained simultaneously by isolation and purification of papaya primary pulp, complete utilization of resources is realized, the preparation steps are simple, the design is reasonable, the operation is simple, the influence on the activity of the enzymes in the preparation process is relatively small, the prepared papain and papaya lipase are high in purity, high in activity and high in enzyme activity recovery rate; and reagents can be recycled after isolation is performed with a strong cation exchange method, so that the reagents are repeated used, the cost is effectively reduced, the preparation can be expanded, and the industrial requirements are met.

Description

technical field [0001] The invention relates to the technical field of biochemical separation, in particular to a high-efficiency preparation method of papaya enzyme preparation. Background technique [0002] Papaya milk contains abundant enzymes such as papain, papain B, chymopapain A, and chymopapain B. [0003] Papain has the characteristics of high temperature resistance, good stability, strong hydrolysis ability, insensitivity to pH changes, metal ions and detergents, etc. It can be used to hydrolyze various proteins, and has been widely used in food, chemical, and textile industries. At the same time, various studies on papain are also increasing day by day, and research, discussion, and seeking the best extraction process have extremely important significance for the development, preparation and application of papain. At present, the traditional methods for extracting papain include: ultrafiltration, salting out, organic solvent, flocculation, affinity chromatography...

Claims

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Application Information

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IPC IPC(8): C12N9/50C12N9/20
CPCC12N9/63C12N9/20C12Y304/22002
Inventor 白新鹏刘亚文黄庆成
Owner HAINAN UNIVERSITY
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