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Method for separating and purifying antibiotic avilamycin for livestock

A technology of avelamycin and veterinary antibiotics, applied in the field of industrial biotechnology and biomedicine, can solve the problems of less research, deviation of analysis results, immature technology, etc., and achieve the effect of simple purification steps and easy operation

Active Publication Date: 2013-04-03
杭州睿岂生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The separation and purification method of avelamycin is mainly completed through several steps such as decolorization, crystallization, and column passing, but at present there are few related studies, and the technology is very immature. There are almost no pure products of avelamycin on the market.
Most of the domestic enterprises and users use the known content of the premix as the reference substance to control production, resulting in large deviations in the analysis results

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Accurately weigh 1.00 g of avelamycin-containing mycelia (20% content) into a 100 mL volumetric flask, and add 30 mL of a mixture of methanol and chloroform at a volume ratio of 1:1. Sonicate for 90 min under the condition that the temperature does not exceed 40°C, dilute to 100 mL, centrifuge at 3000 r / min for 10 min to obtain the supernatant, rehydrate and crystallize according to the ratio of extract: water = 1:1, 4000 r / min Centrifuge for 15 min, and dry at 40°C to obtain the crude extract of avelamycin.

[0030] Weigh the crude avelamycin extract according to the ratio of avelamycin crude extract: silica gel = 1:70 (g:g), dissolve it in 3% 2-propanol chloroform and put it on the column. Elute with a volume of 3% 2-propanol in chloroform at a flow rate of 1 mL / min. Use silica gel HF254 plates (silica gel thickness 0.2 mm) for thin-layer chromatography (TLC) to track the spots, and collect them in segments according to the spot positions. The eluate of the target f...

Embodiment 2

[0034] Accurately weigh 1.00 g of avelamycin-containing mycelium (20%) into a 100 mL volumetric flask, and add 40 mL of a mixture of methanol and chloroform with a volume ratio of 1:1. Sonicate for 100 min under the condition that the temperature does not exceed 40°C, dilute to 100 mL, centrifuge at 3000 r / min for 10 min to obtain the supernatant, rehydrate and crystallize according to the ratio of extract: water = 1:1.5, 4000 r / min Centrifuge for 15 min, and dry at 40°C to obtain the crude extract of avelamycin.

[0035] Weigh the crude avelamycin extract according to the ratio of avelamycin crude extract: silica gel = 1:60 (g:g), dissolve it in 3% 2-propanol chloroform and put it on the column. Elute with a volume of 3% 2-propanol in chloroform at a flow rate of 1 mL / min. Use silica gel HF254 plates (silica gel thickness 0.2 mm) for thin-layer chromatography (TLC) to track the spots, and collect them in segments according to the spot positions. The eluate of the target fra...

Embodiment 3

[0039]Accurately weigh 1.00 g of avelamycin-containing mycelium (20%) into a 100 mL volumetric flask, and add 50 mL of a mixture of methanol and chloroform with a volume ratio of 1:1. Sonicate for 120 min under the condition that the temperature does not exceed 40°C, dilute to 100 mL, centrifuge at 3000 r / min for 10 min to obtain the supernatant, rehydrate and crystallize according to the ratio of extract: water = 1:2, 4000 r / min Centrifuge for 15 min, and dry at 40°C to obtain the crude extract of avelamycin.

[0040] Weigh the crude avelamycin extract according to the ratio of avelamycin crude extract: silica gel = 1:50 (g:g), dissolve it in 3% 2-propanol chloroform and put it on the column. Elute with a volume of 3% 2-propanol in chloroform at a flow rate of 1 mL / min. Use silica gel HF254 plates (silica gel thickness 0.2 mm) for thin-layer chromatography (TLC) to track the spots, and collect them in segments according to the spot positions. The eluate of the target fracti...

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Abstract

The invention relates to the technology of separation and purification, and aims at providing a method for separating and purifying antibiotic avilamycin for livestock. According to the method, organic solvent is adopted to extract mycelium containing avilamycin, then after crystallization, silica column chromatography, ODS column elution separation, recrystallization and leaching, the crystal is rinsed with pure water, and then drying is carried out to obtain pure products of avilamycin A and B. The pure products obtained by the method disclosed by the invention are respectively pure products of avilamycin A and avilamycin B instead of a pure product formed by mixing of various components of avilamycin A; the purifying step is simple and convenient and easy to operate; and the average yield is higher than 37.50%, the purity is higher than 98%, and the pure product can be taken as a reference substance in avilamycin measurement.

Description

technical field [0001] The invention relates to a method for separating and purifying the veterinary antibiotic avelamycin, which belongs to the fields of industrial biotechnology and biomedicine. Background technique [0002] Avilamycin (Avilamycin) is fermented by Streptomyces viridoehrongenes, an oligosaccharide antibiotic belonging to the orthosaccharomycin family. It mainly inhibits Gram-positive bacteria and has poor effect on Gram-negative bacteria. It is a new type of antibacterial and growth-promoting agent. In the livestock and poultry industry, it has attracted more and more attention because of its high safety, no cross-resistance and good and stable growth-promoting effect. Avelamycin is a mixed oligosaccharide antibiotic, currently found to have various structures such as A, B, C, D, F, G, and H. Among them, the most active is the component of avelamycin A, the molecular formula is C 61 h 88 Cl 2 o 32 , the molecular weight is 1403; followed by avelamycin...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07H17/04C07H1/06
Inventor 王宏陈敏潜缓缓
Owner 杭州睿岂生物科技有限公司
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