Preparation method and application of xanthocillin compounds
A technology of compound and crude extract, which is applied in the field of preparation of Xanthocillin compounds, can solve problems such as ineffective treatment of Gram-negative bacterial infections
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Embodiment 1
[0032] 1. The inventor isolated a fungus, Penicilliumchrysogenum Y-S6-1 CCTCC M 2020019 ( figure 1 ), the genomic DNA of the strain was extracted, and the 28S rDNA sequence (its sequence is shown in SEQ ID NO.1) was amplified by polymerase chain reaction (PCR), and a phylogenetic tree was established through sequence comparison. The results showed that the similarity between the bacterium and Penicillium chrysogenum reached 100% ( figure 2 ), indicating that the strain Y-S6-1 belongs to the genus Penicillium, named Penicillium chrysogenum (Penicillium chrysogenum) Y-S6-1, which was preserved in the China Center for Type Culture Collection (CCTCC) on January 7, 2020, Address: Wuhan University, Wuhan, Hubei Province, postcode: 430072, and its deposit number is CCTCC NO: M 2020019.
[0033]2. Inoculate Penicillium chrysogenum (Penicillium chrysogenum) Y-S6-1 CCTCC M 2020019 from the slant preservation test tube on the PDA plate for activation, then pick the spores and inoculate...
Embodiment 2
[0047] Disc Diffusion
[0048] (1) Test sample solution preparation
[0049] Accurately weigh about 2 mg of the ethyl acetate extract of 51 Antarctic-derived microbial fermentation cultures (supernatant + mycelium) (No. 32 is the Penicillium chrysogenum (Penicillium chrysogenum) Y-S6 of Example 1 of the present invention) -1 CCTCCM2020019 crude extract), add DMSO filtered with a 0.22 μm microporous membrane, and configure a 20 mg / ml DMSO solution. The positive control is vancomycin, the concentration is 0.256mg / ml. The negative control was DMSO solvent. The prepared sample solution should be stored in the refrigerator.
[0050] (2) Bacteria-containing test plate preparation
[0051] Inoculate Acinetobacter baumannii, Pseudomonas aeruginosa, Multi-drug resistant Staphylococcus aureus (MRSA), Staphylococcus aureus In Hinton (MH) broth medium, Vibrio alginolyticus (Vibrio alginolyticus) was inoculated in LB medium, and the above indicator bacteria were placed on a constant t...
Embodiment 3
[0056] MIC (minimum inhibitory concentration) method
[0057] (1) Test sample solution preparation
[0058] Accurately weigh about 2 mg each of the pure compounds 1 and 2, and add DMSO filtered through a 0.22 μm microporous membrane to prepare a 2.56 mg / ml solution. The positive control was vancomycin at a concentration of 2.56 mg / ml. The negative control was DMSO solvent. The prepared sample solution should be stored in the refrigerator.
[0059] (2) MIC plate preparation
[0060] The following experiments require aseptic operation in an ultra-clean bench. After overnight incubation the concentration was equivalent to 1.5x10 8CFU / ml indicator bacteria (Acinetobacter baumannii, Escherichia coli, Multi-drug resistant Staphylococcus aureus, MRSA, Staphylococcus aureus, Micrococcus luteus) suspension, diluted with MH broth 1:1000 (OD value 0.04~0.06) as indicator bacteria solution . The operation was performed according to the following steps: (a) MH broth liquid medium wa...
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