Method for extracting purified elastase from pig pancreas

Abstract

The invention relates to a method for extracting and purifying elastase from a porcine pancrea. The method comprises the following steps: the porcine pancreas is minced to form pancreatic chymotrypsin; the pancreatic chymotrypsin is extracted by extract, undergoes plate filtration and ultrafiltration, and is absorbed by ion exchange resins, and eluate is positively and reversely extracted by the reversed micellar extraction method; and the high-purity elastase product is obtained after ultrafiltration, desalinization, concentration and freeze drying of elastase compositions which are reversely extracted. The biological activity of the elastase product prepared by the method reaches more than 1,200 IU / mg, and the specific activity exceeds 2,200 IU / mg . p. As determined by the SDS-polyacrylamide gel electrophoresis method, the molecular weight of the product is about 25,900 Da, and an electrophoresis strip is a single strip and can be taken as a raw material for elastase preparation used in the pharmaceutical industry. The technological line of the method also can be used for extracting and purifying other kinds of protease and biological active molecules from the porcine pancrea.

Description

technical field [0001] The invention relates to a method for extracting and purifying elastase from porcine pancreas, in particular to a method for directly extracting elastase from fresh and frozen porcine pancreas by adopting reverse micellar extraction process. Background technique [0002] Elastase (Elastase), also known as elastase hydrolase, pancreatic elastase, trypsin E and elastase. Elastase is a broad-spectrum proteolytic enzyme characterized by decomposing insoluble elastin. It mainly exists in animal pancreas, skin, aorta, platelets and white blood cells, and the enzyme is also widely distributed in microbial groups. Elastase isolated and purified from animal pancreas is an endopeptidase, consisting of 240 amino acid residues, free of prosthetic groups and metal ions, with a molecular weight of 25900, an isoelectric point (pI) of 9.5, and a white crystal. The crystalline elastase obtained by the conventional separation method is composed of elastase I and elasta...

AI Technical Summary

Problems solved by technology

[0005] Traditional adsorbents often have low adsorption efficiency, strong non-specific adsorption, poor product purity, and heavy subsequent purification burden; in the process of fine-quality purification, it is often difficult to balance yield and purity. For example, molecular sieve chromatography can be used to obtain products with higher purity. However, the activity recovery rate is low, the filler is expensive and the cost is high, and it is not easy to scale up production; in addition, due to the complicated purification process, repeated column purification will further reduce the product yield

In addition, although the use of acetone precipitation and vacuum drying and dehydration can save energy to a certain extent and shorten production time, the use of a large amount of acetone is not conducive to the health of operators, and at the same time, the product activity has a certain loss