Low-temperature alkaline protease gene, engineering bacterium containing same, construction methods of low-temperature alkaline protease gene and engineering bacterium, and low-temperature alkaline pr

Abstract

The invention relates to a low-temperature alkaline protease gene, an engineering bacterium containing same, construction methods of the low-temperature alkaline protease gene and the engineering bacterium, and low-temperature alkaline protease. The sequence of the low-temperature alkaline protease gene is described as SEQ ID No. 2, and the sequence characteristic is 810bp, nucleic acid, single chain, linearity and DNA. The engineering bacterium producing the low-temperature alkaline protease is transformed from a host bacterium bacillussubtilis WB600 and has the characteristic of producing the low-temperature alkaline protease. The construction method of the engineering bacterium comprises the following steps of: obtaining an objective gene, reconstructing the objective gene directionally, constructing an expression carrier and constructing and sieving the engineering bacterium containing the low-temperature alkaline protease gene. The sequence of the mature peptide basic group of the low-temperature alkaline protease is disclosed in SEQ ID No. 2, the optimum operative temperature is 30 DEG C, and the optimum operative pH is 11.0. The optimum temperature of the alkaline protease produced by the engineering bacterium is 30 DEG C, the optimum pH is 11.0, and the invention provides a theoretical basis for the low-temperature alkaline protease added and used in a detergent and has important economic benefit and social benefit.

Description

technical field [0001] The invention belongs to the field of bioengineering and relates to gene mutation and recombination technology, in particular to a low-temperature alkaline protease gene, an engineering bacterium containing the gene, a construction method of the two, and a low-temperature alkaline protease. Background technique [0002] Alkaline protease (EC3.4.21.62) is an enzyme that hydrolyzes protein peptide bonds in the alkaline range, and its optimum pH is generally 9.0-11.0. Used in detergent, food, cosmetics, aquatic feed, tanning, textile and medicine and other fields. [0003] Alkaline protease has a good ability to catalyze the hydrolysis of protein peptide bonds under alkaline conditions, and its alkaline pH is similar to the pH 9.0-12.0 of the environment where clothes are usually washed. Therefore, alkaline protease has been widely used as an additive in washing pharmaceutical industry. The detergent added with alkaline protease has a better decontamina...

AI Technical Summary

Problems solved by technology

The optimum action temperature of Bacillus alkaline protease is 60°C. European laundry is usually soaked in hot water at 40-60°C first, and the alkaline protease in it can fully play its role, while the washing temperature at 40-60°C requires heating of water. It will produce a large waste of energy. In Asia, especially developing countries, it is customary to use tap water for laundry, and alkaline protease cannot exert its maximum enzyme activity at a lower temperature, so that the detergent cannot fully exert its laundry effect. It is necessary to develop an alkaline protease whose optimal temperature is low temperature

[0005] Traditional industrial microbial breeding methods, such as using mutagens to mutagenize enzyme-producing strains, are simple and feasible, but the workload is heavy, and it is difficult to screen ideal strains that meet the requirements of industrial production.

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