Baclicus lincheniformis strain for generating elastase and screening method thereof
A technology of Bacillus licheniformis and elastase, which is applied in the field of bioengineering, can solve the problems of low production safety, limited elastase, insufficient organ resources, etc., and achieves the effects of simple and reliable screening method, easy cultivation and high application value.
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Embodiment 1
[0027] Embodiment 1, the implementation process of a kind of screening method for the Bacillus licheniformis bacterial strain that produces elastase that the present invention proposes is as follows:
[0028] Step 1. Soil sample collection: Take samples from the long-term accumulation of beef, cattle blood vessels, trachea and sewers in cattle and sheep slaughterhouses, or select those pollutants in cattle arteries and blood vessels that are degraded and corrupted by microbial contamination, and use sterilized tweezers Or take the pollutant with a medicine spoon, put it into a sterile empty triangular bottle, and plug it with a cotton plug for later use;
[0029] Step 2. Strain enrichment culture: Weigh 2g of the collected sample, put it into a conical flask filled with 100ml of sterile water, shake it on a shaker at 160r / min for 30min, take 3-4ml of the turbid solution and insert it into a 30ml enrichment flask Medium in a 250ml Erlenmeyer flask, 37°C, 160r / min shaking cultur...
Embodiment approach
[0033] The preferred embodiment of partial condition in above-mentioned step one to step five is as follows:
[0034] Re-screening enzyme activity detection: the above-prepared fermentation broth was centrifuged at 12000r / min for 5min, and the supernatant was retained. The enzyme activity of the liquid was measured as 175-211U / ml;
[0035] Enzyme activity detection plate preparation: 0.3g of elastin, 1.5g of agar powder, 100ml of 0.02mol / L phosphate buffer solution (pH7.5), heated and boiled until the agar melted, and when cooled to 45-50°C, fully shake the Erlenmeyer flask to make the insoluble Mix the elastin evenly, then pour it onto the plate. After the culture medium is solidified, punch a hole on the plate with a hole puncher with a diameter of 6 mm. Take 150 μl of the centrifuged fermentation broth supernatant and add it to the well, and let it stand at 37°C for 8-15 hours. View The size of the hydrolysis circle, the re-screening test plate is as follows figure 2 show...
Embodiment 2
[0038] Embodiment 2: the screening that the present invention proposes is used for the implementation process of the morphological characteristic of the bacillus licheniformis bacterial strain of producing elastase as follows:
[0039] Using the elastase screening method of the present invention, a high-yielding bacterial strain was screened, and the colony was on the common beef extract peptone medium, the colony was off-white, the thalline produced heat-resistant spores, the cysts were not enlarged, Gram staining was positive, and the microscopic It is a single bacterium, straight rod-shaped, blunt and round at both ends, and the spores are elliptical and mobile. The colony is sticky, easy to stir up, and the surface is wet; the specific morphological, physiological and biochemical characteristics of the strain ELAT4 are shown in Table 1; the colony , staining characteristics and physiological and biochemical characteristics are relatively close to Bacillus licheniformis in "...
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