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Manufacturing technique for garamycin B

A technology for the production of gentamicin, which is applied in the field of bacteria producing gentamicin B, can solve the problems of difficult separation, low purity, and many components, and achieve improved purity, high potency, and component ratio high effect

Inactive Publication Date: 2008-08-27
江西制药有限责任公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

It can be seen from the patent that gentamicin B is extracted as one of the by-products (small components) of gentamicin. Due to the large number of components and low content, the separation is difficult, the purity is low, and the cost is high.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0011] Firstly, select and breed the micromonomycin-producing bacteria (Micromonospora echinosporum), and then prepare gentamicin B. The specific steps are as follows:

[0012] 1.1 Preparation of protoplasts:

[0013] Naturally selected micropromycin strain 201-9 from the activated # (Smallomycin component 50%, shake flask titer 1085μ / ml) and 201-65 # (Smallomycin component 55%, shake flask titer 1130μ / ml) was inoculated into mycelial growth medium. The formula of the medium was starch 4.0%, soybean meal 3.5%, glucose 0.5%, corn Powder 2.0%, peptone 0.2%, fish meal 0.2%, ammonium sulfate 0.05%, potassium nitrate 0.01%, calcium carbonate 0.4%, cobalt chloride 8mg / ml, and its pH is 7.7. Shake culture for 60-65 hours, collect the bacteria by centrifugation, wash with 10% sucrose solution, add 15mg / ml lysozyme for enzymolysis in a water bath at 37°C±0.5°C, and microscopically detect the formation of a large number of protoplasts.

[0014] 1.2 Fusion: 201-9 # And 201-65 # The protoplas...

Embodiment 2

[0040] According to the same method as in Example 1, micropromycin strain 201-9 was prepared # And 201-65 # , The formula of the medium in the preparation of protoplasts is starch 3.5%, soybean meal 3.0%, glucose 0.4%, corn flour 1.5%, peptone 0.18%, fish meal 0.3%, ammonium sulfate 0.04%, potassium nitrate 0.01% , Calcium carbonate 0.5%, cobalt chloride 8mg / ml-10mg / ml, its pH is 7.5-7.8; other processes and raw material ratios are the same as in Example 1.

Embodiment 3

[0042] The weight ratio of the culture medium in the preparation of the slant spores of the strain in this example is: bran 2%, soluble starch (C.P) 1.0%, MgSO 4 (C.P)0.05%, KNO 3 (C.P) 0.1, aspartame 0.002%, K 2 HPO 4 (C.P) 0.03%, NaCl (C.P) 0.05%, CaCO 3 (C.P) 0.1%, agar 2.0%, pH adjusted to 7.5. The formula of the medium in the preparation of protoplasts is starch 4%, soybean meal 3.2%, glucose 0.5%, corn meal 2%, peptone 0.18%, fish meal 0.4%, ammonium sulfate 0.05%, potassium nitrate 0.008%, carbonic acid Calcium is 0.35%, cobalt chloride is 8mg / ml-10mg / ml, and its pH is 7.5-7.8; other processes and raw material ratios are the same as in Example 1.

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PUM

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Abstract

The invention discloses a process for producing gentamicin B and a cell protoplast fusion technique is adopted to select a new strain which takes the gentamicin B as a main component through breeding producing strains (micromonospora echinospora) of micronomicinin. The process for producing the gentamicin B which is disclosed by the invention has the advantages that the whole production cycle is shortened, the purity of the gentamicin B is increased, the purity of the gentamicin B which is produced is increased from original 85% to present 90%, the production cost is lowered, and the production capacity of the gentamicin B is extremely increased.

Description

Technical field [0001] The present invention relates to a gentamicin B-producing bacteria, and the present invention relates to a production process of gentamicin B. Background technique [0002] The gentamicin B described in U.S. Patent No. 3091572 1975.10.28 is a mixture of antibiotics obtained by fermentation of Micromonospora crassa and Micromonospora spinosa isolated from by-products of antibiotics. A column chromatography separation method is adopted, that is, the lower layer of a solvent system is adopted, and the solvent system is composed of methanol, chloroform, and ammonium hydroxide as the eluting solvent for separation. Before chromatography, equal volumes of these solvents are mixed and placed to separate the layers. Gentamicin C (consisting of 3 components, C 1 , C 1a And C 2 ) Is eluted first, followed by gentamicin B 1 , Gentamicin X, Gentamicin B, and finally Gentamicin A. It can be seen from the patent that gentamicin B is extracted as one of the by-products (s...

Claims

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Application Information

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IPC IPC(8): C12P19/48C12N15/03C12R1/29
Inventor 黎俊付志纲张威熊兴生张良栋赵鹰英王青吴龙江
Owner 江西制药有限责任公司
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