Herpetosiphon gulosus and application thereof in preparation of medicine for inhibiting methicillin-resistant staphylococcus aureus
A methicillin, staphylococcus-resistant technology, applied in the direction of antibacterial drugs, microorganism-based methods, bacteria, etc., can solve the problems of difficulty in screening new compounds, discovery of new drugs, over-development, etc., and achieve the effect of huge application potential.
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Embodiment 1
[0017] Isolation, purification and identification of embodiment 1 bacterial strain
[0018] The bottom mud sediment was collected from the amaranth field pond in Tangxi Village, Dongli Town, Chenghai District, Shantou City, and air-dried at room temperature. The predatory bacteria in the samples were isolated using the classical method for the isolation of the predatory bacterium Myxobacteria (Escherichia coli induction method). Take 5g of air-dried sediment sample, add 10mL 100μg ml -1 Filter sterilized cycloheximide, soak overnight, then centrifuge and discard the supernatant. With WCX medium (CaCl 2 2H 2 O 1g / L and agar 15g / L, pH 7.2; After the agar is solidified, inoculate Escherichia coli bacterial suspension in the way of streaking) to isolate the predatory bacteria in the sample, and inoculate the sample soaked in cycloheximide in WCX culture The place where the Escherichia coli was streaked was cultured at 30°C. Observe under a stereomicroscope, pick the granular ...
Embodiment 2
[0022] Embodiment 2 preys on the predation of climbing tube fungus 2-13 on MRSA
[0023] Inoculate fresh methicillin-resistant Staphylococcus aureus GDMCC 1.1263 in NA liquid medium (peptone 10g / L, beef extract powder 3g / L, NaCl 5g / L, pH 7.2), and culture overnight at 30°C and 180rpm with shaking. 8000rpm was centrifuged for 5min to collect the thalline, and washed twice with sterile deionized water, and then washed with TPM buffer (Tris-HCl10mM, KH 2 PO 4 1mM,MgSO 4 ·7H 2 (0 8mM, pH 7.6) resuspended cells to make OD 600= 1.0. The predation ability of the predatory Clemania 2-13 on MRSA was evaluated by the direct predation method of flat laying and the confrontation culture predation method respectively. Tiling direct predation method: inoculate 50 μl of resuspended bacteria on TPM solid plate (add 1.5% agar to TPM buffer), dry for later use; inoculate fresh bacteria of predatory climbing tube 2-13 on VY / 2 solid medium for cultivation On the 3rd day, use a puncher to o...
Embodiment 3
[0025] Embodiment 3 preys on the preparation of climbing pipe bacteria liquid fermentation and fermented supernatant crude extract
[0026] The fermentation medium is VY / 4 liquid (yeast 1g / L, CaCl 2 2H 2 O 1g / L, VB 12 0.5mg / L, pH 7.2), add 2% volume ratio macroporous resin XAD-16 before inoculation, and inoculate Herpetosiphongulosus2-13 seed solution according to the volume ratio 2% inoculation amount. 140rpm, shake culture at 30°C for 3 days, collect the resin by filtration with gauze, add 2 times the volume of methanol, shake and extract at 30°C for 1 hour, collect methanol by filtration; repeat the extraction process 3-4 times until the methanol is colorless. Combine the collected methanol on a rotary evaporator at 35°C for 40r min -1 Evaporate to dryness under reduced pressure. A total of 0.96 g of crude extract was obtained by fermenting 2 L of the strain. Dissolve the above crude extract with a small amount of dimethyl sulfoxide (DSMO): water (v / v 1:1), and dilute...
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