Octapeptin as well as preparation and application thereof
A technology of peptides and preparations, applied in the field of new polymyxin analogs, can solve the problems of drug withdrawal and complicated treatment, and achieve the effect of good safety and obvious control effect
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0040] Embodiment 1: Paenibacillus tianmuensis F6-B70 (CCTCC No: M 209149) is fermented according to the following conditions to produce a new compound of formula (I)
[0041] 1. Inoculate the strain F6-B70 on the slant of nutrient agar medium for activation, and culture at 30°C for 1-2 days;
[0042] 2. Put the slant seeds into 50mL seed culture medium in a 250mL Erlenmeyer flask, and culture at 30°C and 200rpm for 24 hours, which is the first-grade seed solution;
[0043] 3. Put the first-grade seed solution into 200mL seed medium in a 500mL Erlenmeyer flask, and shake and culture at 30°C and 200rpm for 24h to obtain the second-grade seed solution;
[0044] 4. The secondary seed solution was inserted into a 2L Erlenmeyer flask containing 500 mL of fermentation medium at an inoculum size of 5-10% (v / v), and cultured for 96 hours at 30° C. with shaking at 200 rpm to obtain a fermentation liquid.
[0045] Nutrient agar medium, prepared as follows: peptone 10g, beef extract 3g,...
Embodiment 2
[0048] Embodiment 2: the separation and purification of formula (I) novel compound
[0049] The fermentation broth obtained in Example 1 can be separated and purified according to the following steps to obtain the pure product of the new compound of formula (I) provided by the present invention.
[0050] 1. Transfer the fermentation broth to a centrifuge bottle, centrifuge at 6500×g for 30 minutes, remove the bacteria, and obtain a clarified fermentation supernatant;
[0051] 2. Pass the supernatant through the Amberlite XAD-16 chromatographic column balanced with water at a constant flow rate, and wash 5 to 10 samples with distilled water and aqueous solution containing 25% (v / v) methanol respectively after loading the samples Bed volume, and then eluted with an aqueous solution containing 80% (v / v) methanol, the elution peak eluted with 80% methanol aqueous solution was collected, concentrated and evaporated to dryness, and recorded as crude product 1.
[0052] 3. Dissolve ...
Embodiment 3
[0054] Embodiment 3: antibacterial activity assay
[0055] 1) Prepare the new compound of formula (I) with a concentration of 256 μg / ml with Mueller Hinton broth medium, and carry out two-fold dilution: the concentrations are respectively 256, 128, 64, 32, 16, 8, 4, 2, 1, 0.5 μg / ml.
[0056] 2) Add 50 μl of the new compound solution of formula (I) at different concentrations (200-0.39 μg / ml) after serial dilution to the 1st to 10th wells of the 96-well polystyrene plate, and add 50 μl Mueller Hinton broth medium (beef extract powder 2g / L, acid hydrolyzed casein 17.5g / L, starch 1.5g / L, solvent is water), as a growth control, add 100μl Mueller Hinton broth medium to the 12th well , as a negative control. Positive controls were polymyxin B and vancomycin.
[0057] 3) After culturing the indicator bacteria for 24 hours, dilute to 10 with Mueller Hinton broth medium 6 cells / ml, add 50 μl of diluted bacterial solution to the 1st to 11th wells, seal and incubate in an incubator a...
PUM
Login to View More Abstract
Description
Claims
Application Information
Login to View More