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Method for improving fermentation level of polymyxin B sulfate

A technology for polymyxin sulfate and fermentation level, which is applied in the field of fermentation formula and fed-feed fermentation process for improving the fermentation level of polymyxin sulfate B, can solve the problems of difficulty in precise control of the fermentation process, fast metabolism, short cultivation period and the like , to achieve the effect of facilitating the utilization of bacteria, promoting synthesis and improving quality

Active Publication Date: 2021-03-26
NCPC NEW DRUG RES & DEV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The main reason is that the polymyxin B-producing bacteria is Bacillus polymyxa, which has fast metabolism and short culture period, and it is difficult to precisely control the fermentation process.

Method used

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  • Method for improving fermentation level of polymyxin B sulfate
  • Method for improving fermentation level of polymyxin B sulfate
  • Method for improving fermentation level of polymyxin B sulfate

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Prepare the medium for fermentation according to the following formula, adopt the preferred medium proportion of the present invention: the basic formula is composed of per liter of medium: wheat flour 140g, amylase 0.11g, sodium chloride 0.8g, magnesium chloride 1.0g, phosphoric acid Potassium dihydrogen 0.7g, calcium carbonate 5g, defoamer 3mL, and the rest is water. The feeding formula is composed of per liter of medium: 23g of peptone, 32g of corn steep liquor, 36g of ammonium sulfate, and the rest is water.

[0039] The fermentation process of the new technology is implemented according to the following specific steps:

[0040] a) Dissolve wheat flour and amylase in an appropriate amount of water, add antifoaming agent, adjust the PH liquid base to 6.92 before elimination, add calcium carbonate, gelatinize at 90°C for 40min, disinfect at 121°C and hold pressure for 30min, the volume after elimination is about 27L . The pH after digestion was measured to be 6.52, ...

Embodiment 2

[0044]On the basis of the process in Example 1, the effects of different formulations and fermentation temperatures on metabolism were investigated. Example 2 The basic formula is the composition of each liter of medium: 150g of wheat flour, 0.12g of amylase, 0.9g of sodium chloride, 0.9g of magnesium chloride, 0.6g of potassium dihydrogen phosphate, 4g of calcium carbonate, 2mL of defoamer, and the rest is water . The feeding formula is composed of per liter of medium: 22g of peptone, 31g of corn steep liquor, 35g of ammonium sulfate, and the rest is water. The difference from the process in Example 1 is that the temperature is controlled at 34°C from the beginning of the fermentation until the dissolved oxygen drops to the lowest value, and the temperature is controlled at 30°C when the dissolved oxygen starts to rise to the end of the fermentation. After 33 hours of fermentation, the microscopic examination of the aging of the bacteria , the pH rose rapidly, and the titer ...

Embodiment 3

[0046] On the basis of the process in Example 1, the effects of different formulations and fermentation temperatures on metabolism were investigated. Example 3 The basic formula is the composition of each liter of medium: 130g of wheat flour, 0.10g of amylase, 0.7g of sodium chloride, 1.1g of magnesium chloride, 0.8g of potassium dihydrogen phosphate, 6g of calcium carbonate, 4mL of defoamer, and the rest is water . The feeding formula is composed of 24 g of peptone, 33 g of corn steep liquor, 37 g of ammonium sulfate, and the rest is water. The difference from the process in Example 1 is: the temperature is controlled at 30°C from the beginning of fermentation until the dissolved oxygen drops to the lowest value, and the temperature is controlled at 26°C when the dissolved oxygen starts to rise to the end of fermentation. When the fermentation is 38 hours, the microscopic examination of the bacterial aging , the pH rose rapidly, and the titer of the fermentation broth reache...

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Abstract

The invention discloses a method for improving the fermentation level of polymyxin B sulfate. The method comprises a fermentation formula and a fed-batch fermentation process, wherein the fermentationformula is divided into a basic formula consisting of a fermentation carbon source and inorganic salt and a fed-batch formula consisting of a fermentation nitrogen source; and the fed-batch fermentation process comprises the following steps: a) performing individual disinfection and sterilization on a fermentation tank; b) performing individual disinfection and sterilization on a fed-batch bottle; and c) supplementing fed-batch liquid consisting of the fermentation nitrogen source at a constant speed in the first and middle stages of fermentation. The fermentation titer of the polymyxin B sulfate obtained by fermentation culture by the method can reach 11000u / mL or above on average, the fermentation titer is improved by 30% or above compared with the original fermentation level, the component proportion of the fermentation liquor is close to the liquid phase chromatogram of a standard substance, and industrial production is easy.

Description

technical field [0001] The invention belongs to the technical field of biological fermentation, in particular to a fermentation formula and fed-batch fermentation process for improving the fermentation level of polymyxin B sulfate. Background technique [0002] Polymyxin is a general term for a family of basic cyclic polypeptide antibiotics composed of various amino acids and fatty acids produced by Bacillus polymyxa. Because different strains produce polymyxins A, B, C, D, E with different chemical structures. The products marketed for clinical application are mainly the sulfate and mesylate salts of polymyxin B and polymyxin E. Among them, the antibacterial effect of polymyxin B is better than that of polymyxin E. Polymyxin has a strong bactericidal effect on Gram-negative bacteria. Due to its relatively large nephrotoxicity, ototoxicity, and neuromuscular blockade, the clinical application of this type of drug is mainly external or topical. Especially for preventing sc...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P21/02C12N1/20C12R1/12
CPCC12P21/02C07K7/62Y02A50/30
Inventor 王昂仲伟潭郭月玲张莉张雪霞高健石英米贯东王崔岩常亮李敏安兰兰彭亮谢新宇
Owner NCPC NEW DRUG RES & DEV
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