Enzyme activity evaluation method for euphausia superba trypsin at low temperature

Abstract

The invention provides an enzyme activity evaluation method for euphausia superba trypsin at a low temperature. The method comprises the following steps: independently obtaining two euphausia superbatrypsin subtypes and a control group of penaeus vannamei trypsin; independently constructing a prokaryotic expression carrier of the euphausia superba trypsin subtypes and a prokaryotic expression carrier of the penaeus vannamei trypsin; independently carrying out extraction, purification and renaturation on the prokaryotic expression carrier of the euphausia superba trypsin subtypes and the prokaryotic expression carrier of the penaeus vannamei trypsin; independently carrying out enzyme activity measurement on the purified euphausia superba trypsin and the penaeus vannamei trypsin; and predicting a positive selection site of an euphausia superba trypsin subtype gene, carrying out mutation, and carrying out enzyme activity measurement. Through a site-directed mutation technology, the functional site of the euphausia superba trypsin is subjected to mutation to verify a position of the site where the euphausia superba trypsin performs a stable catalytic activity at the low temperature, and illustrate a characteristic that the euphausia superba trypsin performs the high catalytic activity at the low temperature.

Description

technical field [0001] The invention belongs to the technical field of marine animal-derived protease function mining, and in particular relates to an enzyme activity evaluation method of Antarctic krill trypsin at low temperature. Background technique [0002] Trypsin is a serine proteolytic enzyme extracted from animal pancreas. It mainly functions in the digestive system of vertebrates and is an essential digestive protease in the intestinal tract of humans and animals. At the same time, trypsin also exists in organisms such as actinomycetes and silkworms. Trypsin plays an important role in the molting of crustaceans and the improvement of immunity. Trypsin from bovine or porcine pancreas has been used as a therapeutic drug in human and animal clinical trials. So far, proteolytic enzymes have been widely used in industry, food, biomedicine and other fields. The field of medicine is mainly used in the following aspects: (1) as an oral drug for some gastrointestinal dise...

AI Technical Summary

Problems solved by technology

[0008] (1) In modern industrial and medical fields, proteases from mammals are mainly used as enzyme preparations. These enzymes can only play the role of larger hydrolytic enzymes at higher temperatures, and the optimum enzyme activity temperature is above 55 ° C, while bacterial contamination It is easy to occur at higher temperature, and if it is applied at low temperature, the enzyme activity will be greatly reduced

[0009] (2) For the field of biomedicine, the antibacterial, antiviral and wound healing effects of traditionally used trypsin preparations are average, and when low temperature is required, the enzyme activity decreases, resulting in increased enzyme usage, increased concentration, and high cost

But it is also difficult to meet the requiremen...

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