Production method of high-purity teicoplanin

A technology for the production of teicoplanin and its production method, which is applied in the field of purification and production of the antibiotic substance teicoplanin, can solve the problems of low production efficiency, hindering the growth of bacterial strains, hindering the formation of teicoplanin, etc., so as to improve production efficiency and realize industrial effect

Active Publication Date: 2008-11-12
ZHEJIANG MEDICINE CO LTD XINCHANG PHAMACEUTICAL FACTORY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In fact, the optimal growth condition for teicoplanin is a weakly alkaline state above pH 7; however, the above separation method can only isolate/purify teicoplanin in an acidic pH culture solution environment, and an acidic environment will hinder the growth of the strain Therefore, the above-mentioned separation/purification method not only has the defect of low production efficiency, but also has the problem of the self-toxicity of the bacterial strain that cannot be solved in the cultivation of the bacterial strain and the isolation process of teicoplanin
The so-called self-toxicity phenomenon refers to that the antibiotic substance-teicoplanin produced by the above-mentioned strains hinders the self-metabolism of the bacterial strain, thereby hindering the production of teicoplanin; for the generation

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] Dissolve 10 g of crude teicoplanin in 200 ml Na 2 CO 3 : NaHCO 3 =0.05M: in the buffer solution of 0.01M, filter to obtain 200ml filtrate, carry out HPLC analysis to this filtrate, the result teicoplanin content is 7.05g, and this filtrate pH value is 9.2, and above-mentioned filtrate is with the flow velocity of 0.2BV / hr Drop on a column (φ3mm×30mm) filled with 200ml Q sepharose gel, and then add 4BV of Na 2 CO3:NaHCO 3 =0.05M:0.1M buffer solution, 4BV of the above buffer solution containing 0.1% Tris was prewashed at a flow rate of 2BY / hr, and finally washed with the above buffer solution containing 1% Tris at 2BV / hr flow rate elution. 400ml of teicoplanin eluent was obtained, and through HPLC analysis, the peak area of ​​teicoplanin A2 in the eluent was 84.6% of the total area, and 6.02g of high-purity crude product was obtained, with a recovery rate of 85.4%.

Embodiment 2

[0041] Adjust the pH of the process liquid obtained by gel chromatography in Example 1 to 7.0, pass through the macroporous adsorption resin JD-1, wash the resin column with 4BV of hydrochloric acid solution at pH=3, and the flow rate is 2BV / hr, and then wash the resin column with 4BV 10% ethanol solution washes with the flow rate of 2BV / hr, then eluted with the flow rate of 1BV / hr with 60% ethanol solution, obtains 320ml eluate, thereby strips out teicoplanin, through HPLC analysis, the The peak area of ​​teicoplanin A2 was 87.2% of the total peak area, and 5.4g of teicoplanin was recovered, the recovery rate was 89.7%

Embodiment 3

[0043] With 300ml of the ethanol eluate in Example 2, adjust the pH to 4.0, add 3g of activated carbon for sugar and 3g of activated carbon for needles, control the temperature at 20-30°C and stir for 30min, then filter, remove the activated carbon to obtain a clarified filtrate.

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Abstract

The invention provides a method for making high-purity teicoplanin. The method comprises the following stages that: stage a, gel chromatography and macropore fractionation by adsorption of a teicoplanin crude product solution are carried out so as to obtain a teicoplanin crude product solution; stage b, active carbon decoloring, ultrafiltration, nanofiltration and crystallization of the teicoplanin crude product solution are carried out so as to obtain high-purity teicoplanin. The method furthest increases the production efficiency of teicoplanin, thereby realizing the industrialization of high-purity teicoplanin.

Description

technical field [0001] The invention relates to a method for purifying and producing the anti-biological substance teicoplanin by using a variant strain actinoplanesteichomyceticus var jianesis; A high-efficiency method for the purification of actinomycetes to produce antibiotic substance-teicoplanin. Background technique [0002] The so-called antibiotics refer to low-molecular compounds that inhibit the reproduction of other organisms in a low-concentration environment below 1 mg / ml. Therefore, the category of antibiotics usually includes not only pharmaceutical antibiotics, but also anticancer agents, agricultural fungicides, herbicides, and insecticides; Chemical structure, etc., can also subdivide the above-mentioned antibiotics. [0003] On the other hand, about 10,000 kinds of antibiotic substances have been isolated from natural products so far; and more than 50% of these antibiotic substances are purified and produced by actinomycetes; most of which actinomycetes ...

Claims

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Application Information

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IPC IPC(8): C07K9/00C07K1/16
Inventor 孙新强葛永红章槐东胡三明
Owner ZHEJIANG MEDICINE CO LTD XINCHANG PHAMACEUTICAL FACTORY
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