Drug-resistant bacteria active substance penicipyrroether A as well as preparation and application thereof
A technology of penicillin ether and active substances, which is applied in the application field of preparing anti-methicillin-resistant Staphylococcus aureus drugs, and can solve the problems of enhanced drug resistance of MRSA
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Problems solved by technology
 Embodiment 1. Isolation and cultivation of Penicillium griseofulvum ZZ380
Get weighed thick-legged crab (Pachygrapsus crassipes) (20.1 grams), soak in 75% ethanol for 10 seconds to remove surface microorganisms, then wash three times with sterile water and then homogenate, take the upper layer liquid after centrifugation to prepare volume The concentration is 1×10 -1 , 1×10 -2 , 1×10 -3 sample solution (take 1mL homogenate and centrifuge the upper layer liquid, add 9mL sterile water to make a volume concentration of 10 -1 sample solution, and diluted step by step to obtain a volume concentration of 10 -2 and 10 -3 sample solution). Take 200 μL of sample solutions of various concentrations and evenly disperse them in a petri dish containing solid medium of potato dextrose agar (PDA, 6 g of potato flour, 20 g of glucose, 20 g of agar, 0.1 g of chloramphenicol, purchased from Hangzhou Microbial Reagent Co., Ltd.). After culturing at 28°C for 5 days, different...
 Example 2. Identification of bacterial species of Penicillium griseofulvum ZZ380
 The species of the obtained strain ZZ380 was identified by ITS rDNA sequence analysis method.
 2.1 Experimental reagents and instruments
 PCR reagent: PrimeSTAR Max DNAPolymerase (TaKaRa), primer (Invitrogen synthesis), primer sequence is:
 Marker: DL2000
 Experimental equipment: centrifuge, electrophoresis instrument, PCR instrument, ABI 3730XL sequencer.
 2.2 Experimental steps
 2.2.1 Extraction of fungal genomic DNA
 Using the Ezup Column Fungal Genomic DNA Extraction Kit (Shenggong), the fungi were ground with liquid nitrogen before use.
 2.2.2 Concentration and quality detection of fungal genomic DNA
 DNA concentration and quality detection was performed using a Nanodrop ultra-micro spectrophotometer.
 2.2.3 PCR amplification
 a. PCR reaction system
 Embodiment 3. Preparation of Penicillium griseofulvum ZZ380 fermentation broth
 Pick Penicillium griseofulvum ZZ380 on the PDA solid slant medium and inoculate it into a 500mL Erlenmeyer flask containing 250mL potato-dextrose broth (PDB, 100g potato, 10g glucose, 35g sea salt, 1L water) liquid medium middle. The culture solution containing the ZZ380 strain was cultivated with rotation (180 rpm) and shaking at 28° C. for 3 days to obtain a strain liquid. Transfer 5 mL of the seed liquid into a 500 mL Erlenmeyer flask containing 250 mL of PDB, and culture it statically at 28°C for 30 days to obtain a fermentation liquid containing the active substance penicillin ether A.
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