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Method for purifying pseudomonas acid A

A technology of pseudomonas acid and desorption liquid, which is applied in antibacterial drugs, pharmaceutical formulations, medical preparations containing active ingredients, etc. The extraction method is cumbersome to operate, etc., to achieve the effect of economical industrial scale production

Inactive Publication Date: 2009-12-02
SHANDONG JIANWEI BIOENG
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The liquid-liquid extraction method is cumbersome to operate and requires a large amount of organic solvents, which is defective from the perspectives of ecology, environmental protection and economy; liquid-solid extraction has some improvements over liquid-liquid extraction, but there are many difficulties in terms of industrial scale

Method used

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  • Method for purifying pseudomonas acid A

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Example 1. Purification of Pseudomonic Acid A

[0038] 1. Adsorb the solution containing pseudomonic acid A with resin and desorb it from the resin

[0039] 1. Preparation of fermented liquid containing pseudomonic acid A:

[0040] Insert Pseudomonas fluorescens P.fluorescens, the producer of Pseudomonas acid A, into a 5L seed tank containing 3.5L sterile seed medium, and then cultivate it under deep aeration and stirring for 10-12h until the cell concentration reaches OD 600nm 4.8-5.2, then under aseptic conditions, this seed culture solution is transferred in the 50L fermentor that 30L aseptic fermentation medium is housed, carries out pseudomonic acid A fermentation culture to 96-110hr, and fermentation terminal The content of Pseudomonas acid A (2300-2600 mg / L) in the fermentation broth was determined by high performance liquid chromatography (HPLC). The HPLC detection conditions mentioned are: chromatographic column: Diamonsil C18 250×4.6mm 5μm; detector: ultravi...

Embodiment 2

[0049] Example 2. Purification of Pseudomonic Acid A

[0050] 1. Adsorb the solution containing pseudomonic acid A with resin and desorb it from the resin

[0051] Pseudomonas is cultured by a known method to obtain a fermentation broth containing pseudomonic acid A. Get 25L of fermented liquid (concentration of Pseudomonas acid A 2510mg / L), adjust pH to 5.0 with 1M HCl, add SP825 resin (processed, 2.5L), wherein, the pretreatment method of SP825 resin is as provided by the manufacturer The instruction manual requires processing. Stir for 30 minutes. Rinse with water to remove residue, pull out the resin with a 60 mesh sieve, pack into a column, and desorb with the method provided in Example 1, wherein the solvent used for desorption is a solvent composed of ethanol+salt+water of 11.9L, and ethanol is contained in the 11.9L solvent 6000ml, 850g of sodium acetate, the rest is water. The ratio of the solvent composed of ethanol+salt+water to pseudomonic acid A was 4ml:21mg. ...

Embodiment 3

[0055] Example 3. Purification of pseudomonic acid A

[0056] 1. Adsorb the solution containing pseudomonic acid A with resin and desorb it from the resin

[0057] Pseudomonas is cultured by a known method to obtain a fermentation broth containing pseudomonic acid A. Take 25L of fermentation broth (concentration of Pseudomonas acid A 2490mg / L), adjust the pH to neutral with 1M NaOH, centrifuge in a high-speed low-temperature centrifuge (centrifugal speed and temperature are 5000rpm and 25°C) for 20 minutes to obtain the clear liquid , and discard the residue after washing with water. The supernatant was adjusted to pH 4.8 with 1M HCl, and passed through a D101 resin (processed, 3.0 L) column for adsorption. After completion, the column was washed with deionized water until the effluent was nearly colorless.

[0058] Analyze with a solvent composed of 9.9L of acetone+salt+water, the 9.9L solvent contains 3000ml of acetone, 300g of sodium chloride, and the rest is water. The...

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Abstract

The invention discloses a method for purifying a pseudomonas acid A, which comprises the following steps: 1) absorption: absorbing solution containing the pseudomonas acid A with resin; 2) desorption: desorbing the pseudomonas acid A from the resin in the step 1) by using a solvent to obtain a desorption solution containing the pseudomonas acid A; and 3) processing the desorption solution containing the pseudomonas acid A obtained by the step 2) to obtain the pseudomonas acid A. The recovery rate and purity of pseudomonas acid A purified by the method provided by the invention reach 79 percent and over 98 percent respectively. The method for purifying the pseudomonas acid A (mupirocin) by using resin of the invention has obvious advantages in aspects of ecology, environmental protection, economy and industrial large-scale production.

Description

technical field [0001] The invention relates to a method for purifying pseudomonic acid A. Background technique [0002] Fermentation with Pseudomonas strains can synthesize multiple chemical components of antibiotics mainly composed of Pseudomonic Acid A. Except for Pseudomonic Acid A, other components are named with letters B-D [E.B.Chain, G.Mellows, J. Chem. Soc. Perkin Trans I. 318 (1977); J.P. Clayton et al., Tetrahedron Lett., 21, 881 (1980); P. J. O. Hanlon, N. H. Rogers, J. Chem. Perkin Trans I. 2665 (1983)]. Pseudomonic acid A is an aminoacyltransaminase inhibitor antibiotic, which can specifically bind to the isoleucine transfer RNA synthetase of bacteria, thereby inhibiting the synthesis of isoleucine-containing proteins in bacteria; Gram-positive bacteria, including Staphylococcus aureus, Staphylococcus epidermidis, Streptococcus pyogenes, and metmoxicillin-resistant bacteria, have high medical value and have been clinically used for a long time. Pirocin ointme...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07D407/06A61K31/351A61P31/04
Inventor 王大然王雅婷乔瑞颖
Owner SHANDONG JIANWEI BIOENG
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