Staphylococcus squirrel cy1-78 and its application in the preparation of fibrinolytic enzyme by fermentation
The technology of Staphylococcus squirrel and fibrinolytic enzyme, which is applied in the biological field, can solve the problems of ineffective oral administration, short half-life and high price, and achieve the effects of simple separation and purification process, easy cultivation and short fermentation period.
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0029] Example 1: Isolation and screening of strains producing fibrinolytic enzyme
[0030] The wild strain isolation and screening method of described Staphylococcus squirrel CY1-78 bacterial strain are as follows:
[0031] (1) Plate separation: cut a silkworm cocoon, take out the silkworm chrysalis, soak it in 75% ethanol aqueous solution at room temperature (about 25°C) for 1min, take it out, cut off the head, open the abdominal cavity and take out the midgut; In a sterile mortar, add 1 mL of sterile saline to grind until homogenized; dilute the homogenate with sterile physiological water to 1×10 4 –1×10 6 times. Draw 0.1mL of 1×10 5 , 1×10 6 , 1×10 7 The 3 times dilution solution was spread on the skim milk plate medium, and cultivated in a 37°C incubator until the number of colonies no longer increased. Pick the colony with a transparent circle around it, streak and inoculate it on a fresh LB plate medium, culture at 37°C until a single colony grows, and then pick a...
Embodiment 2
[0044] Example 2: Classification and identification of enzyme-producing strains
[0045] The culture characteristics of the CY1 strain are as follows: the strain was inoculated on an LB plate, and after culturing at 37°C for 24 hours, milky white colonies could be seen, the colonies were opaque, with neat edges and a smooth surface (see the attached photo for the colony photos). figure 1 ). The CY1 strain was inoculated on the skimmed milk plate medium, and after culturing at 37°C for 24 hours, a large transparent circle was formed around the colony (see the attached photo for the colony photo). figure 2 ).
[0046] The cell characteristics of the CY1 strain are as follows: Gram-positive, spherical, and 0.8-1.2 μm in diameter. Bacteria are often distributed singly or in irregular clusters, and the optical microscope photos after Gram staining are shown in the appendix image 3 .
[0047] The total DNA of the CY1 strain was extracted, and the 16S rDNA fragment of the strai...
Embodiment 3
[0049] Mutation breeding of embodiment 3 Staphylococcus squirrel
[0050] The CY1 strain of Staphylococcus squirrel was subjected to ultraviolet mutagenesis, and the CY1-78 strain of Staphylococcus squirrel was obtained by screening.
[0051] (1) Cell preparation: 20 mL of LB liquid medium was inserted into slanted cells of Staphylococcus squirrel CY1 strain, and cultured at 37° C. and 200 r / min shaking for 12 h. Take 1mL of the bacterial solution in a centrifuge tube, centrifuge at 5000r / min for 5min, discard the supernatant, add 1mL of sterile saline to resuspend the bacterial cells, and centrifuge again to collect the bacterial cells. Resuspend the bacteria in normal saline in a 100mL Erlenmeyer flask for later use. The composition and preparation method of the LB liquid medium are the same as in Example 1.
[0052] (2) Ultraviolet mutagenesis: Take 6 pairs of sterile petri dishes with a diameter of 6 cm, and add 1 mL of the above bacterial suspension respectively. Open ...
PUM
Login to View More Abstract
Description
Claims
Application Information
Login to View More