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Antibiotic, and preparation method and application thereof

A technology for the activation of antibiotics and strains, which is applied to the preparation methods of peptides, antibacterial drugs, chemical instruments and methods, etc., can solve the problems of high price, low potency, purity and stability, and achieve high titer and good inhibition. Antibacterial effect, avoid the effect of low activity

Active Publication Date: 2015-10-28
SHANTOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The purpose of the present invention is to provide an antibiotic and its preparation method to solve the shortage of separation and purification process in the production of nisin, high price, low potency, purity and stability, etc.

Method used

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  • Antibiotic, and preparation method and application thereof
  • Antibiotic, and preparation method and application thereof
  • Antibiotic, and preparation method and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0032] Preparation of Antibiotics

[0033] (1) Preparation of Lactococcus lactis fermentation broth: Prepare the medium according to the formula in Table 1. After sterilizing at 115°C for 30 minutes, dissolve the purchased strain Lactococcus lactis ATCC 11454 dry powder with 0.5mL liquid medium, and then add 5% The ratio of (v / v) was added to 5 mL of liquid medium, and activated for 24 hours in a constant temperature incubator at 30°C. After the strains are activated, pour 25 mL of solid medium into the plate, use an inoculation needle to inoculate a little activated bacterial liquid on the plate, and incubate at 30°C for 18 hours. After the growth of the strains, pick a single colony and inoculate it in 30 mL of seed liquid medium, and cultivate it in a constant temperature incubator at 30 °C for 20 h. Then the cultured seed solution was inoculated into fresh 200 mL liquid medium at a ratio of 3% (v / v) for fermentation and culture, and the culture condition was 30° C. for 16...

Embodiment 2

[0039] Preparation of Antibiotics

[0040](1) Preparation of Lactococcus lactis fermentation broth: Prepare the medium according to the formula in Table 1. After sterilizing at 115°C for 30 minutes, dissolve the purchased strain Lactococcus lactis ATCC 11454 dry powder with 0.5mL liquid medium, and then add 5% The ratio of (v / v) was added to 5 mL of liquid medium, and activated for 24 hours in a constant temperature incubator at 29°C. After the strains are activated, pour 25 mL of solid medium into the plate, use an inoculation needle to inoculate a little activated bacterial liquid on the plate, and incubate at 29°C for 18 hours. After the growth of the strains, a single colony was picked and inoculated in 30 mL of seed liquid medium, and cultivated in a constant temperature incubator at 29 °C for 20 h. Then the cultured seed solution was inoculated into fresh 200 mL liquid medium at a ratio of 3% (v / v) for fermentation and culture, and the culture condition was 29° C. for 1...

Embodiment 3

[0044] Preparation of Antibiotics

[0045] (1) Preparation of Lactococcus lactis fermentation broth: Prepare the medium according to the formula in Table 1. After sterilizing at 115°C for 30 minutes, dissolve the purchased strain Lactococcus lactis ATCC 11454 dry powder with 0.5mL liquid medium, and then add 5% The ratio of (v / v) was added to 5 mL of liquid medium, and activated for 26 hours in a constant temperature incubator at 31°C. After the strains are activated, pour 25 mL of solid medium into the plate, use an inoculation needle to take a little activated bacterial liquid and inoculate it on the plate, and incubate at 31°C for 20 hours. After the growth of the strains, a single colony was picked and inoculated in 30 mL of seed liquid medium, and cultured in a constant temperature incubator at 31 °C for 22 h. Then the cultured seed solution was inoculated into fresh 200 mL liquid medium at a ratio of 3% (v / v) for fermentation and culture, and the culture condition was 3...

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Abstract

The invention relates to an antibiotic, and a preparation method and an application thereof. The preparation method comprises the following steps: activating Lactococcal lactis ATCC 11454, picking single colony, culturing, centrifuging to remove bacterial strains to obtain a fermentation broth, adjusting the pH value of the fermentation broth to 2, carrying out rotary evaporating concentration, adjusting the pH value of the obtained concentrate to an initial value, centrifuging, taking the obtained supernatant, and filtering to obtain a fermentation concentrate; and mixing the fermentation concentrate with a PBS buffering solution, adding a double water phase system formed by Triton X-114 and MgSO4, centrifuging to realize phase balance of double water phases, taking the obtained upper phase, and diluting the upper phase by using the PBS buffering solution to prepare the antibiotic containing nisin and Triton X-114. The antibiotic has a substantial bacteriostasis effect on Gram-positive bacteria. The antibiotic includes nisin and Triton X-114, and the combination of the nisin and the Triton X-114 makes the antibiotic have high titer. The preparation method has the advantages of simple extraction process and low cost, and is suitable for intermittent and large-scale production and processing of high-tilter and high-yield antibiotics.

Description

technical field [0001] The invention relates to the technical field of deep processing of natural products, in particular to an antibiotic and its preparation method and application. Background technique [0002] Nisin, also known as nisin, is a polypeptide substance produced by lactic acid streptococcus. It is a bacterial antibacterial peptide. It consists of only 34 amino acids and has a molecular weight of about 3510Da. Its active molecules often form dimers and tetramers. form exists. Nisin consists of a hydrophilic end and a hydrophobic end, and is a typical surface active substance. Nisin has the ability to inhibit most Gram-positive bacteria, and the spore-forming Gram-positive bacteria are particularly sensitive to Nisin. In general, Nisin has no inhibitory effect on Gram-negative bacteria, yeast and mold. Therefore, it is widely used in the food industry as a food preservative. After eating, it is quickly hydrolyzed into amino acids under the physiological pH co...

Claims

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Application Information

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IPC IPC(8): C12P21/02C07K14/315C07K1/14A61K38/16A61P31/04
Inventor 刘杨刘黎玲李夏云肖湘
Owner SHANTOU UNIV
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