Purification method for papain

Abstract

The invention discloses a purification method for papain. The purification method comprises the following steps: (1) adding water into pawpaw pulp to obtain a pawpaw pulp solution; (2) adding a compound enzyme Extyme-A; (3) carrying out plate frame filtering, namely carrying out plate frame filtering on the pawpaw pulp solution treated by the step (2); (4) centrifuging, namely carrying out centrifugal sedimentation on a filtering solution and collecting a centrifuging solution; (5) adding a compound enzyme Extyme-B; (6) coarsely filtering, namely filtering the centrifuging solution by a polyvinylidene fluoride filtering membrane; (7) finely filtering, namely filtering a coarsely-filtered material solution by a polysulfone ultra-filtration membrane, and concentrating to obtain a papain concentrated solution; and (8) drying in vacuum to obtain the papain. According to the purification method for the papain, the filtering effect can be improved obviously, so that coarse filtration and ultra-filtration are smooth, the filtering time is shortened, and the working efficiency and the enzyme activity yield are improved.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for purifying papain. Background technique [0002] Papain is a class of thiol proteases. Immature green papaya fruit is rich in content, the enzyme has high activity, good thermal stability, and has the characteristics of natural, hygienic, and safe. It is widely used in food, medicine, cosmetics, leather, and textile fields, and the demand and price are increasing year by year. . [0003] At present, the commonly used extraction methods for the production of papain include: 1. drying or spray drying, 2. flocculation, 3. salting out, 4. ultrafiltration refining. [0004] The process described in method 1 is relatively simple and primitive, and its disadvantages are that high temperature greatly damages the enzyme activity, the yield of enzyme activity is low, and there are many impurities, and the obtained enzyme activity is about 1 million / g. [0005] The flocculation m...

AI Technical Summary

Problems solved by technology

The problem with this method is that because the papaya pulp contains more colloids, lipids, and starch substances, the dissolution and release of the papaya enzyme in the stirring and extraction process is not complete, and there are more residues in the subsequent plate and frame filtration, and the filtrate is unclear; The membrane pore size is very small, which is easy to cause blockage of the filter membrane, and the filtration speed is slow, which affects the filtration and separation of papain. The production process must be shut down to clean the filter membrane, resulting in the loss of a certain amount of papain and reducing the yield of enzyme activity.

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