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Preparation method of low-temperature protease

A low-temperature protease and protease technology, applied in the field of low-temperature protease preparation, can solve the problems of low enzyme activity, poor decontamination effect, energy waste, etc., achieve high enzyme activity, save energy, and enhance the effect of low-temperature washing effect

Inactive Publication Date: 2017-05-10
SUZHOU KUANGSHI JUNCHI BIOLOGICAL SCI & TECH
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Problems solved by technology

[0002] At present, the protease added to the detergent is derived from microorganisms at normal temperature, and its enzyme activity is very low at low temperature, so it can only play a good role in removing protein pollutants in high temperature seasons such as summer
These normal temperature proteases have very poor decontamination effect in low temperature environments such as winter; to increase the decontamination effect, the water temperature needs to be raised to 30-40 degrees, resulting in a waste of energy

Method used

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  • Preparation method of low-temperature protease

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Embodiment

[0015] Preparation of Protease from Psychrophilic Bacteria Colwellia psychrerythraea

[0016] The first step is to design and synthesize the protease gene of the psychrophilic bacterium Colwellia psychrerythraea, and insert it into the pET28 expression vector to construct the recombinant expression plasmid of DNA ligase. The N-terminus of recombinant psychrophilic bacterium Colwellia psychrerythraea protease has 6 consecutive histidine affinity purification tags derived from pET28 vector, which is used for immobilized nickel ion affinity chromatography purification.

[0017] The second step is to recombinantly express the protease of psychrophilic bacteria Colwellia psychrerythraea. The psychrophilic bacterium Colwellia psychrerythraea protease recombinant expression plasmid was transformed into Escherichia coli expression host BL21 (DE3), and the psychrophilic bacterium Colwellia psychrerythraea protease recombinant expression strain was obtained. The expression strain was c...

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Abstract

The invention discloses a preparation method of low-temperature protease. The preparation method comprises the following steps: 1) designing and synthesizing a Colwellia psychrerythraea protease gene; 2) recombining and expressing Colwellia psychrerythraea protease; 3) performing affinity purification on the Colwellia psychrerythraea protease; 4) detecting the activity temperature dependency of the Colwellia psychrerythraea protease. The optimal temperature range of the enzymatic activity of the prepared low-temperature protease is 8 to 30 degrees; in the temperature range, various proteins can be dehydrated, so that the protease is very suitable for serving as protease of a cold water detergent.

Description

technical field [0001] The invention belongs to the technical field of genetic engineering, and in particular relates to a preparation method of low-temperature protease. Background technique [0002] At present, the proteases added to detergents are derived from normal temperature microorganisms, and their enzyme activity is very low at low temperatures, so they can only play a good role in removing protein pollutants in summer and other high temperature seasons. These normal temperature proteases have very poor decontamination effect in low temperature environments such as winter; to increase the decontamination effect, the water temperature needs to be raised to 30-40 degrees, resulting in a waste of energy. Contents of the invention [0003] The object of the present invention is to address the defects of proteases added to existing detergents, and develop a protease that is active at low temperatures. Within the range, proteolytic activity can be effectively exerted....

Claims

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Application Information

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IPC IPC(8): C12N15/70C12N9/52
CPCC12N9/52
Inventor 刘喜朋
Owner SUZHOU KUANGSHI JUNCHI BIOLOGICAL SCI & TECH
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