Neutral proteinase gene constructed by molecular chaperone prsA, protein, bacillus subtilis, preparation and application

Abstract

The invention belongs to preparation of a new gene and construction of an engineering bacterium, and particularly relates to a neutral proteinase gene constructed by a molecular chaperone prsA, a protein, bacillus subtilis, preparation and an application. The molecular chaperone PrsA and the neutral proteinase gene are integrated to construct pHT43-npr-PrsA recombinant plasmid by a molecular biology technology and finally converted into a host bacterium, namely the bacillus subtilis WB800N, to construct a new bacillus subtilis engineering bacterium CGMCC No. (China General Microbiological Culture Collection Center number) 14837; the molecular chaperone PrsA and the neutral proteinase gene are co-expressed; and an expression level of neutral proteinase is effectively increased.

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 prsA 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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