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Screening and application of a probiotic Bacillus licheniformis with high production of compound enzyme

The technology of Bacillus licheniformis and strains, applied in the field of agricultural microbiology, can solve the problems of being susceptible to the influence of internal and external environments, high cost of enzyme preparation preparation, poor growth promoting effect, etc., and achieves fast growth and reproduction, obvious bacteriostatic effect, The effect of strong resistance

Active Publication Date: 2020-05-29
湖北华大瑞尔科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The purpose of the present invention is to improve the poor growth-promoting effect of a single probiotic, the disadvantages of high enzyme preparation cost, unstable properties, and easy to be affected by internal and external environments, and provide a screening and application of a high-yield compound enzyme probiotic Bacillus licheniformis

Method used

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  • Screening and application of a probiotic Bacillus licheniformis with high production of compound enzyme
  • Screening and application of a probiotic Bacillus licheniformis with high production of compound enzyme
  • Screening and application of a probiotic Bacillus licheniformis with high production of compound enzyme

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Effect test

Embodiment 1

[0031] Example 1: Isolation and identification of bacterial strains

[0032] 1. Isolation of strains

[0033] The isolate samples were taken from the feces of 90-day-old healthy broiler chickens in a chicken farm in Wuhan City, Hubei Province. After sample collection, pick up a little feces with a sterile cotton swab and put them in a 7mL EP tube filled with 3mL sterile water. After bathing in 80°C water for 10 minutes, spread the bacterial solution on the LB solid medium. The dirty white colonies with pleats and irregular edges are used for pure culture, and the pure culture is morphologically observed by Gram staining for preliminary screening. The bacteria are long rod-shaped, single, paired or chain-shaped Gram-positive bacilli, After 20 hours, spores were formed, and nearly mesogenic ellipsoid spores were produced, and the cysts were slightly enlarged. The bacterial strain of this invention was named Bacillus licheniformis HDRaBLp, and was preserved in subculture. For t...

Embodiment 2

[0061] Embodiment 2: the bacteriostasis performance verification of bacterial strain

[0062] When preparing indicator bacteria (enteropathogenic Escherichia coli, Staphylococcus aureus, Clostridium welchii) suspension, pick a small amount of indicator bacteria from the inclined surface with an inoculation needle, and inoculate them into conventional nutrient broth medium respectively, 37 Cultivate for 24 hours at ℃, adjust the concentration of the bacterial suspension to about 10 9 CFU / mL.

[0063] Using the method of plate diffusion (Lyver et al., 1998; Lin Dong et al., 2001), dip a small amount of Bacillus licheniformis into the center of the poured nutrient agar plate with an inoculation stick, culture at 37°C for 24 hours, and then place the plate upside down on the Cover the petri dish with a thin layer of chloroform and fumigate for about 30 minutes to kill living cells and allow colonies to fix on the surface of the plate, then remove the petri dish cover and let the ...

Embodiment 3

[0066] Embodiment 3: Stress resistance and growth characteristics of Bacillus licheniformis strain HDRaBLp

[0067] 1. Tolerance test

[0068] (1) Bile salt tolerance test

[0069] After 2 generations of activation of Bacillus licheniformis strain HDRaBLp (isolated from 90-day-old healthy broiler manure in a chicken farm in Wuhan, Hubei Province in December 2013), 1 mL was inoculated into 9 mL containing 0%, 0.15%, 0.30% , in conventional LB liquid medium with 0.50% bile salts, cultivated at 37°C for 12 hours, diluted to an appropriate multiple, took 0.1 mL and spread it on a solid LB plate, observed and counted the growth, and repeated 3 times for each treatment. The results showed that HDRaBLp could tolerate 0.15%, 0.30%, and 0.50% bile salt concentrations (see Table 4). The concentration of bile in the digestive tract fluctuates in the range of 0.03% to 0.3%, therefore, Bacillus licheniformis strain HDRaBLp can tolerate the intestinal bile salt environment.

[0070] Tabl...

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Abstract

The invention belongs to the technical field of agricultural microorganism application, and in particular relates to the screening and application of a probiotic bacillus licheniformis with high production of complex enzymes. The bacillus licheniformis of the invention has obvious antibacterial effect on common livestock and poultry enteropathogenic bacteria such as escherichia coli, staphylococcus aureus and Clostridium welchii. The invention includes the isolation and identification of the bacillus licheniformis strain, the research on biological characteristics and its application as a feed additive for livestock and poultry, and the like. The bacterial strain of the present invention produces cellulase, amylase and protease, is resistant to low pH value of gastric acid and high osmotic pressure of intestinal bile salt, has fast growth rate, strong stress resistance, strong antibacterial property and strong enzyme production ability , can be used to prepare microbial additives for livestock and poultry feed.

Description

technical field [0001] The invention belongs to the technical field of agricultural microbiology, and specifically relates to the screening and application of a probiotic Bacillus licheniformis with high-yield compound enzymes. The bacillus licheniformis is effective against common intestinal pathogenic bacteria such as E. Staphylococcus and Clostridium welchii have obvious antibacterial effect. The invention includes the isolation and identification of the bacillus licheniformis strain, the research on the biological characteristics and its application as a feed additive for livestock and poultry. Background technique [0002] The use of antibiotic feed additives was once considered to be one of the greatest biotechnologies in animal husbandry production in the 20th century. It has played a huge role in preventing and controlling animal diseases, improving livestock and poultry production efficiency, and meeting the needs of human animal products. However, with the passage...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20A23K10/18C12R1/10
CPCA23K10/18C12N1/205C12R2001/10
Inventor 王喜亮李越章恺伦金卉肖运才周祖涛石德时金秀娥
Owner 湖北华大瑞尔科技有限公司
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