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Preparation method for gentamicin C1a

A technology of gentamicin and seeds, which is applied in the field of preparation of gentamicin C1a, can solve the problems of cumbersome operation and high production cost, and achieve the effect of stable output and low production cost

Inactive Publication Date: 2017-03-15
WUXI FORTUNE PHARMA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] The purpose of the present invention is to disclose a preparation method of gentamicin C1a, in order to overcome the defects of high production cost and cumbersome operation in the preparation method in the prior art

Method used

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  • Preparation method for gentamicin C1a

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Experimental program
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Effect test

Embodiment 1

[0029] The preparation method of gentamicin C1a comprises the following steps.

[0030] Step (1), carry out microwave mutagenesis with gentamicin B (the mutant of Micromonospora aculeatus) producing bacterium, draw 2mL spore liquid and be suspended in EP tube, EP tube is placed in ice-bath beaker, in Treat in a microwave oven at 700W, 2450MHz for 10s; then use a chemical mutagen for chemical mutagenesis, the chemical mutagen is nitrosoguanidine, and the concentration of the mutagen is 0.4mg / mL, at 20°C, in a constant temperature incubator Constant temperature cultivation was carried out for 6 h. Gentamicin C1a-producing strains were screened.

[0031] Step (2), selecting stable and relatively high-yielding gentamicin C1a-producing bacteria, and performing slant culture in slant medium. The components in the slant culture medium are (unit: g / L): cornstarch 1.8, KNO 3 0.05, NaCl0.04, MgSO 4 0.03,K 2 HPO 4 0.02, wheat flour 1.8, calcium carbonate 0.09, agar 1.7, and the bal...

Embodiment 2

[0040] The preparation method of gentamicin C1a comprises the following steps.

[0041] Step (1), carry out microwave mutagenesis to gentamicin B producing bacteria, draw 2mL spore liquid and suspend in EP tube, put EP tube in ice bath beaker, process 30s with 700W, 2450MHz in microwave oven; Then use A chemical mutagen is used for chemical mutagenesis, the chemical mutagen is nitrosoguanidine, the mutagen is used at a concentration of 0.7 mg / mL, and a constant temperature culture is carried out in a constant temperature incubator at 30° C. for 7 hours. Gentamicin C1a-producing strains were screened.

[0042] Step (2), selecting stable and relatively high-yielding gentamicin C1a producing bacteria, and carrying out slant culture. The components in the slant medium are (unit: g / L): corn starch 2.0, KNO 3 0.1, NaCl0.05, MgSO 4 0.05,K 2 HPO 4 0.03, wheat flour 2.0, calcium carbonate 0.10, agar 2.0, and the balance is water. The pH of the slant medium was natural (pH7±0.2). ...

Embodiment 3

[0051] The preparation method of gentamicin C1a comprises the following steps.

[0052] Step (1), carry out microwave mutagenesis to gentamicin B producing bacteria, draw 2mL spore liquid and suspend in EP tube, put EP tube in ice-bath beaker, process 60s with 700W, 2450MHz in microwave oven; Then use A chemical mutagen was used for chemical mutagenesis, the chemical mutagen was nitrosoguanidine, and the concentration of the mutagen was 1.0 mg / mL, and the culture was carried out in a constant temperature incubator at 38° C. for 8 hours. Gentamicin C1a-producing strains were screened.

[0053] Step (2), select stable and relatively high-yielding gentamicin C1a producing bacteria, carry out slant culture, the components in the slant medium are (unit: g / L): cornstarch 2.2, KNO 3 0.15, NaCl0.06, MgSO 4 0.07,K 2 HPO 4 0.04, wheat flour 2.2, calcium carbonate 0.11, agar 2.3, and the balance is water. The pH of the slant medium was natural (pH7±0.2). Cultured at 40°C for 15 day...

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Abstract

The invention provides a preparation method for gentamicin C1a. The preparation method comprises the following steps: performing the microwave mutagenesis and chemical mutagenesis to gentamicin B producing bacteria, selecting a new bacterial strain in which the gentamicin C1a is the main component, performing the microwave mutagenesis firstly and performing the chemical mutagenesis secondly to obtain the producing bacteria of the gentamicin C1a, performing the fermenting cultivation, separating the obtained fermentation liquor through a micro-filtration membrane, an ultra-filtration membrane and a nano-filtration membrane so as to obtain a crude product of the gentamicin C1a. The yield of the gentamicin C1a is very stable through the continuous passage of the producing bacteria of the obtained gentamicin C1a, the production cost is lower, and the preparation method is suitable for the industrialization application.

Description

technical field [0001] The invention relates to the technical field of biopharmaceuticals, in particular to a preparation method of gentamicin C1a. Background technique [0002] Gentamicin is an aminoglycoside antibiotic mainly used to treat bacterial infections, especially those caused by Gram-negative bacteria. Gentamicin can bind to the 30s subunit of bacterial ribosomes and block bacterial protein synthesis. [0003] "Pharmacopoeia of the People's Republic of China" 2000 edition stipulates that the C components of gentamicin include C1, C1a, C2a, and C2, and among them, gentamicin C1a can be used as the parent body of the synthetic antibacterial drug emetycin. The molecular structural formula of gentamicin C1a is as follows: [0004] [0005] Gentamicin C1a is the precursor for the synthesis of etimicin, and etimicin sulfate is a semi-synthetic aminoglycoside antibiotic with high efficiency, low toxicity and resistance to drug-resistant bacteria developed by my coun...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P19/48C12N15/01C12N13/00C12R1/29
CPCC12P19/485C12N13/00C12N15/01
Inventor 杨亮付静曹峥游云龙赵永俊金丹甜蔡云洁
Owner WUXI FORTUNE PHARMA
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