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Application and Application Method of Bacillus Cereus Keratinase

A technology of Bacillus cereus and keratinase, applied in the direction of microorganism-based methods, chemical instruments and methods, biochemical equipment and methods, etc.

Active Publication Date: 2021-08-20
BIOTECH RES CENT SHANDONG ACADEMY OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] However, the above-mentioned article also discloses that the optimum action temperature of keratinase is mainly concentrated at 40-70°C, such as 45°C for Streptomyces and 60°C for Nocardiae; Keratinase has high stability at 50°C and 60°C, the enzyme activity is still 100% after 3 hours of treatment at 50°C, and the half-life is 2.5 hours at 60°C

Method used

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  • Application and Application Method of Bacillus Cereus Keratinase
  • Application and Application Method of Bacillus Cereus Keratinase
  • Application and Application Method of Bacillus Cereus Keratinase

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Embodiment 1

[0021] The method for obtaining Bacillus cereusY-15 keratinase and the method for measuring keratinase activity are the same as those in the patent document CN 107868762A;

[0022] The present invention detects the enzymatic properties of the keratinase such as the optimum enzyme activity temperature and the stability at different temperatures, as follows:

[0023] 1.1 Optimum enzyme activity temperature: use 2% keratin as the substrate, dilute the enzyme solution to an appropriate concentration with pH 9.0 Tris-HCl buffer, mix with the substrate 1:1, and measure the enzyme activity at pH 9.0. The reaction temperature range is 25-70°C, measure a point every 5°C, and take the highest enzyme activity as 100%;

[0024] 1.2 Stability at different temperatures: Take an appropriate amount of diluted enzyme solution and incubate at 40-70°C for different times, then use 2% keratin as a substrate, and measure the residual enzyme in the enzyme solution after incubation at pH 9.0 live. ...

Embodiment 2

[0047] Keratinase of the present invention is applied in removing protein stain, and concrete method is as follows:

[0048] Tolerance of keratinase in detergent: Dilute keratinase to an appropriate concentration, mix it with different brands of detergents with a mass fraction of 2% 1:1, and after incubation at room temperature for 12 hours, use the Folin-phenol method to measure the residual enzyme live. The enzyme activity of keratinase not mixed with detergent was taken as 100% when it was incubated for 1 hour.

[0049] The washing effect of keratinase on protein stains: the concentration of Ariel detergent is 1%, the concentration of added enzyme is 10 U / ml, and the cotton cloth stained with stains (ketchup, ginger powder, sweet bean sauce, hot sauce, blood stains) is cut to an appropriate size. Washing conditions: 30ºC, 150 rpm for 1 h. The washing effect of Ariel detergent without enzyme solution was used as a control.

[0050] The enzyme activity of keratinase remain...

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Abstract

The invention belongs to the technical field of enzyme application, and in particular relates to an application of Bacillus cereus Y‑15 keratinase. The key protection of the present invention is the application of Bacillus cereus Y-15 keratinase in the preparation of cleaning products, especially the application of the keratinase in the preparation of washing products. The keratinase of the present invention is obtained by inoculating B. cereus Y-15 seed liquid into an optimized enzyme-producing medium and culturing it; it has excellent thermal stability and meets the following requirements: the half-life of the enzyme activity at 40°C is 34-36.5 h; the enzyme half-life t1 / 2 at 60°C is 5.5-6h, the half-life at 65°C is 2.2-2.5h, and the half-life at 70°C is 0.58-0.65h. In addition, the keratinase of the invention can tolerate reducing agents, oxidizing agents and detergents, and has potential application value in industrial production.

Description

technical field [0001] The invention belongs to the technical field of enzyme application, and in particular relates to an application of bacillus cereus keratinase. Background technique [0002] Wang Qiuying and others disclosed in the article "Application Research Progress of Keratinase" that keratinase has a wide range of application fields, mainly including fertilizer, feed, food and medicine, detergent, leather and other industries. [0003] However, the above-mentioned article also discloses that the optimum action temperature of keratinase is mainly concentrated at 40-70°C, such as 45°C for Streptomyces and 60°C for Nocardiae; Keratinase has high stability at 50°C and 60°C, the enzyme activity is still 100% after 3 hours of treatment at 50°C, and the half-life is 2.5 hours at 60°C. [0004] Most of the reported keratinases belong to the serine protease S8 family, which can be inhibited by PMSF. At present, there is an urgent need to find keratinase resources with mo...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N9/54C11D3/386C12R1/085
CPCC11D3/38618C12N9/54
Inventor 边斐岳寿松夏晗陈高马德源于金慧张燕
Owner BIOTECH RES CENT SHANDONG ACADEMY OF AGRI SCI
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