Serratia marcescens ll-413 strain and its application in preparing serrapeptase
A technology of Serratia marcescens and LL-413, which is applied in the application field of preparing serrapeptase, can solve the problems of unseen fermentation process research, high price, few production enterprises, etc., and achieve separation The effect of simple purification process, easy cultivation and short fermentation period
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Embodiment 1
[0029] Example 1: Isolation and Screening of Rapeptase Producing Strain
[0030] The isolation and screening methods of described LL-413 strains are as follows:
[0031] (1) Preliminary screen screening: Take 1 silkworm cocoon (3.1g), peel off the cocoon shell, take out the silkworm chrysalis (1.6g), soak it in 75% ethanol aqueous solution by volume for 15s, take it out, and carefully cut off the head with tweezers and scissors Partially, cut open the abdominal cavity and take out the internal organs; put the internal organs into a sterile mortar, add 1 mL of sterile deionized water and grind to a homogenate; dilute the homogenate with sterile water to a gradient of 10 -1 -10 -6 times. Pipette 0.2 mL of each dilution solution, spread it onto skim milk plate medium, and cultivate in a 30°C incubator until the number of colonies no longer increases. Pick the colonies that are red and form a transparent circle on the plate, streak and inoculate on fresh nutrient agar plate med...
Embodiment 2
[0050] Example 2: Classification and identification of LL-413 strain
[0051] The LL-413 strain was inoculated on a nutrient agar plate and cultured at 30°C for 24 hours. Bright red colonies were visible on the front, with neat edges and a smooth and shiny surface; the back was also bright red; after culturing at 37°C for 24 hours, pale red colonies were visible. Pink colony, the colony surface is smooth, the edge is neat, and the back is light yellow; there is a slight odor. The LL-413 strain was inoculated on the skim milk plate medium, and after culturing at 30°C for 24 hours, bright red colonies and clear transparent circles around the colonies can be seen, and the colony photos are attached. figure 2 .
[0052] The bacterial characteristics of the LL-413 strain are as follows: Gram-negative, short rod-shaped, about 2.0-2.5 μm in length, 0.8-1.0 μm in width, without flagella and capsule, and the photos of the bacteria are attached image 3 .
[0053] The total DNA of t...
Embodiment 3
[0056] Example 3: Method for preparing serrapeptase by LL-413 strain shake flask fermentation under initial conditions
[0057] Taking the LL-413 strain as the enzyme-producing strain, under the 50mL shake flask fermentation scale, a seed expansion culture step was added, and the method for preparing serrapeptase by shake flask fermentation was as follows:
[0058] (1) The LL-413 strain was inoculated into the slant medium, and cultured at 30°C for 36 hours to obtain the activated slant of the LL-413 strain; the final concentration of the slant medium was composed of: peptone 10g / L, beef extract 3g / L , NaCl 5g / L, agar 20g / L, the solvent is water, and the initial pH is 7.0. The culture medium was heated until the agar melted and then packed in test tubes, sterilized by high pressure steam at 121°C for 15min, and then placed on the inclined plane;
[0059] (2) Inoculate the thalline of the LL-413 strain after the activation and cultivation in step (1) into 50 mL of seed medium,...
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