Preparation process of high-activity papain

Abstract

The invention discloses a preparation process of high-activity papain. The preparation process comprises the following operation steps that (1) fresh chaenomeles speciosa is cleanly cleaned; peels and seeds are removed; the chaenomeles speciosa is cut into small blocks; distilled water is added; pulping is performed; (2) complex enzyme (cellulase and pectinase) is added and is uniformly stirred; enzymolysis is performed; (3) suction filtration is performed to obtain fresh pawpaw juice; (4) the fresh pawpaw juice is centrifuged to obtain supernate; (5) ethyl alcohol and an enzyme activity protection agent (L-cys and EDTA (ethylene diamine tetraacetic acid)) are added into the supernate; the PH is regulated to be 6.0 to 8.0; still standing is performed to obtain a mixed solution; the mixed solution is centrifuged; precipitates are taken; freeze drying is performed until the constant weight is reached; a papain crude product is obtained. The processes of juice squeezing, compound enzymatic hydrolysis, enzyme activity protection agent addition and the like are used; the content of protease in the pawpaw juice is obviously increased. Compared with an ordinary enzyme preparation extracting method, the preparation process has the advantages that the enzyme activity of the papain reaches 15.5 thousand U / g; through application test, the crude product of the papain can obviously improve the tenderness of a meat product.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a preparation process of high-activity papain. Background technique [0002] Xuan papaya is the ripe fruit of the crabapple, belonging to the shrub of the genus Papaya in the family Rosaceae. It is a famous food and drug specialty in Anhui. Xuan papaya contains a lot of trace elements and nutrients and there are many types. It has the title of "Baiyi Fruit". Xuan papaya fresh weight contains about 1.1% crude protein, which also has structural protein and biological function protein, such as a large number of proteases, rich in a large amount of essential amino acids needed by the human body. In terms of application, the protease in papaya is extracted and purified, and then re-produced into products such as meat tenderizer powder, feed additive, biscuit loosening agent, beer clarifying agent and hydrolyzed animal protein series. [0003] At present, the preparation tech...

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Problems solved by technology

[0004] The salting-out precipitation method changes the solubility of the protein by changing the concentration of neutral inorganic salts, and at the same time causes damage to the hydration film on the surface of the protein to cause protein precipitation. The working conditions of this process are relatively mild and can retain Enzyme activity, but more salting-out solvents will remain in the precipitate, which is not conducive to subsequent purification

[0005] Ultrafiltration is an advanced extraction technology, which is the direction of papain refining. The problem with this process is that because the papaya slurry contains more colloids and starch substances, the dissolution and release of papain in the stirring extraction process is not complete. There are many filter residues in the plate and frame, and the filtrate is unclear; in the membrane separation and filtration stage, due to the small pore size of the filter membrane, the filter membrane will be clogged, the filtration speed is slow, and the separation of papain will be affected. loss of enzyme activity

[0006] The ethanol precipitation method is a common method for the preliminary extraction and purification of papain. Ethanol extraction of papain has simple materials, convenient operation, and high resolution. It does not need to use some processes to remove the extractant in the precipitation. Vacuum filtration can be used, but the ethanol method will reduce the enzyme activity of protease during the extraction process

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