Oceanobacillus sp. XC22919 and application thereof
A technology of XC22919 and bacillus, which is applied to the application field of Pacific bacillus and its secondary metabolites in the field of quorum sensing inhibition, can solve problems such as difficulty in eradicating Pseudomonas aeruginosa, and achieve the effect of simple and easy-to-control preparation process and simple conditions.
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Embodiment 1
[0039] Example 1: Isolation, purification and identification of marine bacteria
[0040] 1. Strain screening
[0041] The 10 sea mud samples were collected from different depths in the Pacific seamount, and the 10 samples were placed in sterile sample bags and stored at -20°C. The 10 sea mud samples were subjected to the following operations to screen bacteria.
[0042] (1) Take 5g of spare sea mud, add an appropriate amount of sterile seawater in a sterile mortar, grind it into fine particles, place it in a conical flask pre-filled with sterilized LB liquid medium, and culture it with shaking at 30°C and 180rpm , enriched for 12 hours to obtain enriched sea mud suspension.
[0043] (2) Then take 1ml of the enriched sea mud suspension and carry out gradient dilution with sterile deionized water to obtain 10 -1 -10 -9 Nine dilutions of sea mud suspension.
[0044] (3) Take 100 μL of the sea mud suspension of each dilution and spread it on the LB plate, culture it in a cons...
Embodiment 2
[0072] Example 2: Growth Inhibition and Violet Inhibition of Violet Bacillus ATCC12472 by Pacibacterium XC22919
[0073] 1. The effect of different concentrations of Pacific bacillus XC22919 extract on the purple pigment production of C.violaceum ATCC12472
[0074] 1.1 C.violaceum ATCC12472 bacteria solution and culture medium preparation
[0075] 1.1.1 Pick a single colony of Violet Bacillus ATCC12472 from the plate and inoculate it in a pre-prepared conical flask (250ml conical flask, 100ml medium) filled with LB liquid medium, cultivate overnight at 30°C, 180rpm to logarithm During the period, the violaceum ATCC12472 bacterial liquid was obtained for future use.
[0076] 1.1.2 Use 27 conical vials of 25ml, and divide them into 3 groups in total, which are blank group, negative control group and experimental group, among which 7 concentration gradients are set in the experimental group, and each group has 3 parallel samples. Put 10ml LB liquid culture medium in the vial, ste...
Embodiment 3
[0091] Example 3: Pyocyanin inhibition and cluster motility inhibition of wild-type Pseudomonas aeruginosa by Pacibacterium XC22919
[0092] 1. Pyocyanin inhibition of wild-type Pseudomonas aeruginosa by Pacific bacillus XC22919
[0093] 1.1.1 Inoculate wild-type Pseudomonas aeruginosa in PB liquid medium, grow to the logarithmic phase at 37°C, and divide 27 25ml Erlenmeyer vials into 3 groups, namely blank group and negative control group , the experimental group, wherein the experimental group is set with 7 concentration gradients, and each group has 3 parallel samples, and 10ml of PB liquid medium is put in each conical vial, sterilized by high temperature and high pressure, and set aside.
[0094] 1.1.2 Dilute Pseudomonas aeruginosa bacteria solution with fresh PB liquid medium, adjust the concentration of the bacteria solution to OD≈0.05, and add it to the conical vials of each experimental group and negative control group (except the blank group). 10ml of bacterial solu...
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