Neutral protease gene built by molecular chaperone DnaK, protein, bacillus subtilis, and preparation and application of neutral protease gene built by molecular chaperone

Abstract

The invention belongs to new gene preparation and engineering bacterium building, and particularly relates to a neutral protease gene built by a molecular chaperone DnaK, protein, bacillus subtilis, and preparation and application of the neutral protease gene built by the molecular chaperone DnaK. The molecular biology technology is used for integrating the molecular chaperone DnaK with the neutral protease gene; the pHT43-npr-DnaK recombinant plasmids are built and are finally converted into a host bacterium of bacillus subtilis WB800N; a novel bacillus subtilis engineering strain CGMCC (China General Microbiological Culture Collection Center) No.14836 is built, so that the molecular chaperone DnaK and the neutral protease gene realize the coexpression; the expression level of the neutralprotease is effectively improved.

Description

technical field [0001] The invention belongs to the preparation of new genes and the construction of engineering bacteria, in particular to a neutral protease gene, protein, bacillus subtilis constructed by using molecular chaperone DnaK and its preparation and application. Background technique [0002] The traditional tanning process will produce a lot of pollution. The four characteristic wastes discharged by tanning are: wastewater, sludge, waste gas and solid waste. Among them, the discharge of all kinds of pollution is the traditional ash-alkali hair removal process in the water field. Although the hair removal effect of the ash-alkali hair removal method is good, the dissolved hair, epidermis, and ash-alkali and other pollutants account for the total amount of tanning wastewater. 40% of pollutants. The cleaning and depilatory processes studied in recent years mainly include enzymatic hair removal, hair-preserving hair removal based on ash-alkali method, organic amine ...

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Problems solved by technology

However, the above-mentioned methods all have drawbacks. It is easy to form inclusion bodies when transformed into Escherichia coli, and it is necessary to break the wall to obtain enzymes, which will not only reduce enzyme activity but also limit the secretion and expression of enzymes; transformation into Pichia pastoris requires methanol to Induce enzyme production, but methanol is easy to evaporate, difficult to detect concentration, repeated addition is not only cumbersome to operate, but also prone to pollution, in addition, it will cause fire hazards in the presence of large amounts of methanol, and is not suitable for protein production such as food

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